- Research
- Open access
- Published:
Potential antidiabetic activity of probiotic and Garcinia kola-yoghurt and its role in regulation of male fertility-stimulating hormones in high-fat diet/low dose streptozotocin-treated rats
Food Production, Processing and Nutrition volume 6, Article number: 55 (2024)
Abstract
Garcinia kola Heckel seed is widely used in the African traditional medicine as a aphrodisiac and male fertility enhancer. Probiotics can reestablish glucose homeostasis and improve blood lipid profiles by altering the composition of the intestinal flora. The study was planned to assess the efficacy of co-administration of Garcinia kola seed aqueous infusion and probiotic yoghurt in the management of diabetes and associated male fertility-stimulating hormones abnormalities. G. kola seed infusion was prepared, assessed for radical scavenging capacity, total phenolic content and phenolic profile using HPLC. Fermented yoghurt was prepared and inoculated with probiotic mixture. Rats were given a high-fat diet for four weeks and received an intraperitoneal injection of streptozotocin (30 mg/kg) to induce type 2 diabetes. Diabetic rats were received 200 mg/kg freeze-dried infusion of G. kola seed, probiotics yoghurt, and probiotic yoghurt mixed with G. kola once a day for four weeks. The levels of glucose, insulin, testosterone, follicle-stimulating hormone, luteinizing hormone, inflammatory indicators, oxidative markers, lipid profiles and liver as well as kidney biochemical indicators were measured. The administration of G. kola seed, probiotic yoghurt, or their combination to diabetic rats demonstrated potential anti-diabetic effects as evidenced by the down-regulation in glucose, insulin, lipid profile, oxidative markers, and inflammatory markers simultaneously with an up-regulation in testosterone, FSH, and LH levels compared to diabetic rats. G. kola seed, probiotic yoghurt, or their combination increased testosterone, FSH, and LH levels and are thought to have therapeutic promise for T2DM and its related oxidative stress.
Graphical Abstract
Introduction
By 2045, there will be 629 million cases of type 2 diabetes mellitus (T2DM) worldwide according to the prediction of the International Diabetes Federation (Cho et al. 2018). T2DM, a complex disorder that affects the host’s homeostasis, is characterized by high fasting blood glucose, decreased insulin efficacy, abnormal carbohydrate and lipid metabolism, and mild gut microbial dysbiosis (Al Bataineh et al. 2023). The risk of vascular inflammation, major micro- and macro-vascular changes, and multiorgan dysfunction which can result in a wide range of diabetes complications, including neuropathy, nephropathy, retinopathy, and cardiovascular disorders is, in fact, increased by persistent hyperglycemia)Mansour et al. 2023). It has been determined that 25% to 50% of men with T2DM have low testosterone (T) levels, resulting in male hypogonadism or erectile dysfunction (Van Cauwenberghe et al. 2022). Testosterone, follicle-stimulating hormone (FSH) and luteinizing hormone (LH) are the major fertility-stimulating hormones in males. Testosterone stimulates and maintains spermatogenesis and fertility in conjunction with FSH. LH regulates testosterone production by Leydig cells, which are found in the interstitium of the testis (Oduwole et al. 2021). Insulin resistance or insufficient insulin levels can have an adverse effect on the brain, pituitary gland, gonads, and perigonads. As a result, T2DM is associated with the reduced secretion of testosterone, FSH, LH, and gonadal steroids such gonadotropin-releasing hormone (Andlib et al. 2023). The astonishing rise in the prevalence of type 2 diabetes around the world, as well as its economic cost, highlights the need for new strategies to halt the disease’s onset and progression, especially since currently available diabetes reducers have side effects such as hypoglycemia and digestive tract problems (Mohamed & Abdel-Salam 2021). Additionally, there hasn’t been medications that addresses diabetics’ reproductive health in male up to this point (Andlib et al. 2023).
Human health is impacted by the gut microbiota’s balance. Diabetes may be facilitated by an imbalance in the intestinal flora. However, dietary interventions can prevent diabetes by restoring normal intestinal flora (Wang et al. 2023). Probiotics, live microorganisms that have health benefits when consumed in appropriate amounts, can improve reestablish glucose homeostasis and blood lipid profiles by altering the composition of the intestinal flora. In addition, probiotics can generate short-chain fatty acids (SCFAs), which can improve gut flora and suppress pathogenic ones, lowering the incidence of type 2 diabetes mellitus (Luo et al. 2023).
Garcinia kola Heckel seeds from the African rain forest (Malpighiales order, Clasiaceae family, Bull. Soc. Bot. France 30 (Rev. Bibliogr.): 150 (1883), theplantlist.org, website accessed on August 29, 2023) are known as “Onie” (in the Fang-Beti languages of Cameroon, Gabon, and Equatorial Guinea), “Orogbo” (in the Yoruba language of Nigeria), or even male kola or bitter-kola (Farahna et al. 2017) because of their bitter flavour. In West African traditional medicine, these seeds are used to treat a variety of ailments, such as diabetes mellitus (Adedara et al. 2015) and malaria (Ogunkunle et al. 2014). A growing number of research (Adedara et al. 2015; Farahna et al. 2017; Idris et al. 2020) indicate that G. kola seeds have anti-inflammatory and anti-diabetic properties. The seed is used in folk medicine to treat fever, digestive, inflammatory, brain, and liver issues. It is also used as an aphrodisiac and a fertility-boosting substance. The major phytochemicals present in G. kola seeds are flavonoids, saponins, tannins, phenols, glycosides, and alkaloids (Kareem et al. 2022).
It has been determined that probiotics hypoglycemic mechanism is connected to the control of the gut microbiota. Consequently, it is thought that taking the aphrodisiac and fertility enhancer G. kola with probiotics would be more efficient in management of diabetes and its associated problems, particularly the decrease in male fertility-stimulating hormones. Therefore, the study was planned to assess the role of supplementation with G. kola, probiotic yoghurt or their mixture in regulation of blood glucose and male fertility-stimulating hormones (testosterone, FSH and LH) levels in a T2DM rat model.
Materials and methods
Materials
Garcinia kola Heckel seeds were purchased from a market in Saudi Arabia. Botanists from Qassim University College of Agriculture verified the seeds. G. kola seeds were peeled, dried, grounded and stored until use. Fresh cow milk samples was obtained from healthy lactating animals in local farm, Cairo, Egypt. Cow milk is available in Arab countries, and G. kola seeds can be imported from the Kingdom of Saudi Arabia or from Nigeria. Probiotic starter cultures of DVS-ABT2 (containing Streptococcus thermophilus, Lactobacillus acidophilus and Bifidobacterium bifidum) were imported from Chr. Hansen’s Lab., Copenhagen Denmark. Chemicals and pure reagents were imported from Sigma (Sigma-Aldrich, St. Louis, MO, USA). Elisa kits were imported from Sunlong (Sunlong Co., Ltd. China).
Animals
Adult male Sprague Dawley rats that were 4 weeks old and weighed 111.36 ± 8.01 g as Mean ± SD were used. The National Research Centre’s animal house in Cairo, Egypt, supplied the animals. Individually housed in stainless steel metabolic cages with free access to food and water, the animals were maintained at room temperature (24 ± 2 °C and 40–60% relative humidity) with 12-h light and dark cycles. The study was approved by the National Research Center’s Medical Research Ethics Committee (MREC), in accordance with the requirements of the pertinent Egyptian legal framework, the Helsinki Declaration, the institutional Animal Care and Use Committee’s (IACUC) guidelines and recommendations, and WHO regulations governing the ethical conduct of scientific research. The No of Ethical Approval Certificate is 24911122022.
Methods
Preparation of aqueous infusion of G. kola (GK)
According to Farahna et al. (2017) and Abdel-salam et al. (2009 and 2018), fifty grams of G. kola seeds powder were placed in a beaker, 500 ml of distilled water (70 °C) was poured over the plant material, then the beaker was closed and the material infused for 6 h, then the mixture was filtered twice, first through cheese-cloth (50% cotton, 50% polyester) and then through filter paper (Whatman No.2). The amount of the obtained infusion was preserved in sterile dark bottles in a cool environment (4° C) until further use. The infusion of G. kola was freeze dried by freeze drier (Crest Alpha 1–4 LSC plus Germany). Later, the freeze-dried powder was sieved by 140 meshes then kept, in polyethylene pouch bags, in refrigerator until used.
High-performance liquid chromatography (HPLC) fingerprinting of GK aqueous infusion
An Agilent 1260 series was used for the HPLC analysis. 4.6 mm × 250 mm i.d., 5 μm, Eclipse C18 column was used for the separation. Water (A) and 0.05% trifluoroacetic acid in acetonitrile (B) were the components of the mobile phase, which had a flow rate of 0.9 ml/min. The linear gradient was sequentially programmed into the mobile phase as follows:0 min (82% A); 0–5 min (80% A); 5–8 min (60% A); 8–12 min (60% A); 12–15 min (82% A); 15–16 min (82% A) and 16–20 (82%A). At 280 nm, the multi-wavelength detector was monitored. For each of the sample solutions, 5 l of injection volume was used. The column was kept at a constant temperature of 40 °C. The retention times of the identified compounds of interest were measured. To create a curve showing peak area and retention time in a chromatogram, the extract was injected into a high-performance liquid chromatographic device. The concentration of the sample is then determined by comparing the peak area of the sample with that of the standard relative to the standard’s concentration.
Determination of total phenolic content of GK aqueous infusion
Using the Folin-Ciocalteu reagent, the total phenolic content of freeze-dried GK infusion was colorimetrically assessed (Singleton et al. 1999). Gallic acid equivalents (GAE), reported as mg/g of sample, were used to measure total phenolic content. The assay was performed in triplicate, and the mean and standard deviation are shown for the outcomes.
Determination of antioxidant activity of GK aqueous infusion
GK infusion’s capacity to scavenge free radicals was assessed using three methods DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging assay, ABTS (2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonate) radical cation assay and FRAP (ferric ion reducing antioxidant power). The DPPH radical scavenging activity was assessed according to the method of Nguyen et al. (2017). To 1 ml of the freeze-dried GK infusion in ethanol with varied concentrations (25, 50, and 75 µg/ml), 0.1 mM of DPPH was added. After fully vortexing, the reaction mixture was kept at room temperature for 30 min. At 517 nm, the absorbance was spectrophotometrically quantified. The assay was performed in triplicate. The following equation was used to calculate the % DPPH scavenging activity:
The ABTS radical cation assay was performed as described by Prior et al., (2005). The ABTS+ cation radical was generated by mixing 10 mg of ABTS and 2 mg potassium persulfate in water and storing it in the dark at room temperature for 12–16 h before use. After diluting the ABTS+ solution (1 ml) with methanol (60 ml), the absorbance was measured at 734 nm, 30 min after the addition of 5 μl of each freeze-dried GK infusion concentration (25, 50, and 75 µg/ml) to 3.995 ml of diluted ABTS+ solution. The assay was performed in triplicate. The following equation was used to calculate the % ABTS scavenging activity:
The FRAP assay was performed as described by Gomez et al. (2023). The components 300 mM acetate buffer, 10 ml tripiridyltriazine in 40 mM hydrochloric acid, and 20 mM ferric chloride (FeCl36H2O) were combined in a ratio of 10:1:1 at 37°C to create the FRAP reagent. 5 μl of each freeze-dried GK infusion concentration (25, 50, and 75 µg/ml) was combined with 3.995 ml of FRAP reagent, and thoroughly mixed. The reduction of ferric tripiridyltriazine to ferrous tripiridyltriazine form in the reaction mixture produced a complex with a strong blue colour. After 30 min of incubation at 37°C, the absorbance was measured at 593 nm against a reagent blank made up of 3.995 ml of FRAP reagent and 5 μl of distilled water. The assay was performed in triplicate. Plotting the absorbance at 593 nm allowed for the creation of a calibration curve using various FeSO4 concentrations.
Preparation of fermented probiotics-yoghurt (Zabady)
According to the procedures outlined by Tamime and Robinson (1999) and Abdel-Salam et al. (2018), probiotic fermented yoghurt was prepared. Whey protein powder was added to milk samples to standardize them, and then the milk was pasteurized for 15 min at 85 °C before being cooled to 40 °C. Streptococcus thermophilus, Lactobacillus acidophilus, and Bifidobacterium bifidum probiotic cultures were inoculated and incubated for 4 to 8 h at 42 °C.
Preparation of diets for animals
A regular, balanced diet with a total kcal value of 412 kcal/kg (12% protein, 10% corn oil, 10% sucrose, 58.5% maize starch, 5% fibre, 3.5% AIN-93 salt mixture, and 1% AIN-93 vitamin mixture) was prepared according to Reeves et al. (1993). A high-fat diet (HFD) with a total kcal value of 528 kcal/kg (12% protein source, 30% lard, 1% cholesterol, 10% sucrose, 42.5% maize starch, 3.5% salt mixture, and 1% vitamin mixture) was prepared according to Zhu et al. (2017). Each diet, with a known weight, was added to crockery inside the metabolite cages on a daily basis.
Type 2 diabetes induction
According to the method of Srinivasan et al. (2005) and Qian et al. (2015), type 2 diabetes was induced in rats. Thirty rats were divided into normal group (n = 6) and HFD group (n = 24). Rats in the normal group were given the balanced diet, while rats in the HFD group were given the HFD for four weeks until the abdominal circumference increased by 7 cm compared to the normal group rats (Novelli et al. 2007), then the HFD group’s rats were injected with streptozotocin (STZ; Sigma-Aldrich, St. Louis, MO). Each rat received an intraperitoneal injection of a single low dosage of STZ (30 mg/kg, dissolved in 0.1 M sodium citrate buffer at pH 4.4). After 72 h, a blood glucose metre (One Touch, LifeScan, Zug, Switzerland) was used to measure the animal’s fasting blood glucose (FBG). The high fasting blood glucose level (over 200 mg/dl) in rats who had diabetes was evidence that the disease had started.
Groups and administrations
The non-diabetic group was considered as the normal control (NC) group and maintained on the balanced diet. The twenty four diabetic rats were continued to feed on the HFD for further 4 weeks after dividing them into four groups (6 rats in each group depending on Mead’s resource equation which is often used for estimating sample sizes of laboratory animal experiments) as follows; diabetic control (DC) group where diabetic rats were untreated, G. kola (GK) group where diabetic rats orally administrated with freeze-dried G. kola aqueous infusion (200 mg/kg, dissolved in distilled water) for 4 weeks, probiotics-yoghurt (PY) group where diabetic rats orally administrated 1 ml of probiotic yoghurt containing 108 CFU of probiotics for 4 weeks, and Gk + PY group where diabetic rats orally administrated 1 ml of probiotic yoghurt (containing 108 CFU of probiotics) mixed with freeze-dried G. kola aqueous infusion (200 mg/kg rat body wight) for 4 weeks.
The oral dose of 200 mg/kg of G. kola infusion was used in this study based on studies that estimated the daily doses of G. kola used in traditional medicine to be between approximately 100 and 300 mg/kg (orally) (Adedara et al. 2015; Ogunkunle et al. 2014), as well as taking into account the outcomes of Ajayi et al. (2011) study who reported that 200 mg/kg of G. kola aqueous extract exhibited neuro-protective effect in mice while Cetkovic-Cvrlje et al. (2022) found that 100 mg/kg of G. kola aqueous extract had no improvement effect on type 1 diabetes in mice.
Thus, the experiment lasted approximately 8 weeks, during which time the total food intake, body weight gain and feed efficiency ratio (Body weight gain/total food intake) were calculated. Blood samples were collected from slightly anaesthetized rats after overnight fast over heparin and without heparin. The blood samples were centrifuged at 3500 rpm for 15 min to separate plasma and serum which stored at -80 ºC till analysis. Finally, rats in the study were euthanized by decapitation and dissected to obtain kidney and liver.
Biochemical analysis
Serum of each rat was analyzed for insulin, testosterone (T), luteinizing hormone (LH), C-reactive protein (CRP), interlukin-6 (IL-6), tumor necrosis factor (TNF-α) and transforming growth factor β1 (TGF-β1) using sandwich ELISA detection kits (SinoGeneclon Biotech Co., Ltd.), Fasting blood glucose levels was determined according to Trinder (1969). The activities of alkaline phosphatase (ALP), aspartate transaminase (AST) and alanine transaminase (ALT) were determined according to Rheinhold and Seligron (1953) and Reitman and Frankel (1957), respectively. The levels of albumin, creatinine and urea and the activity of lactate dehydrogenase (LDH) were determined depending on Doumas et al. (1997), Larsen (1972) and Fawcett and Scott (1960) and Zimmerman and Weinstein (1956), respectively. Plasma total cholesterol (T-Ch), high-density lipoprotein cholesterol (HDL-Ch), low-density lipoprotein cholesterol (LDL-Ch), and triglycerides (TG) were determined according to Watson (1960), Burstein et al., (1970), Schriewer et al., (1984) and Megraw et al. (1979), respectively. Cholesterol/HDL-C ratio was also calculated. Red blood cells lipid peroxidation (MDA) glutathione peoxidase (GPx), nitric oxide (NO) and superoxide dismutase (SOD) activity were determined according to Ohkawa et al. (1979), Paglia and Valentine (1967), Montgomery and Dymock (1961) and Nishikimi et al. (1972), respectively.
Statistical analysis
Statistical analyses were done using SPSS version 21. The results were expressed as mean ± standard error (SE) and analyzed statistically using one-way analysis of variance (ANOVA) Duncan test was used to analyze the statistical differences between groups. The results of body weight gain of normal and diabetic rats post feeding on the high fat diet for 4 weeks were statistically analyzed using T-test. The statistical significance of difference was taken at P ≤ 0.05.
Results
HPLC fingerprinting analysis of GK aqueous infusion
The HPLC detected 18 known phenolic compounds in the freeze-dried G. kola infusion. The presence of different flavonoids and phenolic components in the GK freeze-dried infusion was demonstrated by the HPLC chromatogram (Fig. 1). These phytochemicals were examined from the chromatogram in significant amounts and are shown in Table 1. The most prevalent phenolic component in GK was querectin (103955.7 g/g), which was followed by hesperetin (5913.21 μg/g), daidzein (4504.99 μg/g), and naringenin (345.45 μg/g). Coffeic acid (3.71 μg/g) was the least prevalent substance.
The HPLC chromatogram of GK aqueous infusion
Total phenolic content and antioxidant activity of GK aqueous infusion
Table 2 displays the total phenolic content (146.4 mg/g) of the freeze-dried GK infusion. Owing to antioxidant molecules in GK namely phenolic compounds, it exhibited radical scavenging activity as revealed in Table 2. The DPPH radical scavenging, ABTS radical cation and FRAP assays were used to determined the antioxidant activity of the freeze-dried GK infusion. At low concentration (25 μl/ml), GK scavenged 52.93 and 75.53% of the DPPH and ABTS radicals, respectively and recorded 78.00% FRAP.
Bioassays in a T2DM rat model
Effects of GK, probiotics-yoghurt and their combine on the growth performance
Figure 2 depicts the body weight gain of HFD and normal rats after four weeks of consuming a high-fat diet. Compared to the NC group, the high fat diet considerably accelerated the body’s weight gain. Table 3 displays the effects of G. kola, fermented probiotics, and their combination on the development and performance indices of diabetic rats. Despite the fact that there was no discernible difference in the food intake among the various groups after the 4-week experiment, diabetic rats showed a significant (p < 0.05) weight loss as compared to the NC group. In comparison to the DC group, final body weight of the G. kola, fermented probiotics-yoghurt, and their combined groups significantly (p < 0.05) increased.
Body weight gain of normal (n = 6) and HFD (n = 24) rats post feeding on HFD (4 weeks). #, significantly different from the normal rats at P < 0.05
Effects of GK, probiotics-yoghurt and their combine on glucose, insulin and inflammatory markers
Figure 3 shows serum FBG, insulin and inflammatory markers levels of experimental rats. FBG of DM group (231.17 mg/dl) was significantly higher than NC group (94.07 mg/dl). Compared to NC group (4.82 mU/L), DM group was at significantly higher insulin concentration (7.83 mU/L). Inflammatory markers (TGF-β1, TNF-α, CRP and IL-6) of DM group (67.27 ng/ml, 17.87 pg/ml, 5.57 ng/ml and 75.68 pg/ml, respectively) were significantly higher than NC group (24.42 ng/ml, 6.45 pg/ml, 2.63 ng/ml and 25.92 pg/ml, respectively). However, FBG, insulin and inflammatory markers of G. kola, fermented probiotics-yoghurt and their combine groups were significantly (p < 0.05) less than DM group. The co-administration with G. kola and fermented probiotics-yoghurt revealed the highest improvement in FBG, TGF-β1, TNF-α, CRP and IL-6 levels (138.34 mg/dl, 35.98 ng/ml, 8.95 pg/ml, 3.05 ng/ml and 28.66 pg/ml, respectively).
Effects of GK, probiotics-yoghurt and their combine on FBG (A), insulin (B), TGF-β1 (C), TNF-α (D), CRP (E) and IL-6 (F). Values are set as mean ± SE (n = 6). At a statistical significance level of P < 0.05, various superscript letters were added to represent group differences
Effects of GK, probiotics-yoghurt and their combine on the male fertility-stimulating hormones and oxidative markers
Compared to NC group, all diabetic rats (Fig. 4) showed significant decrease in testosterone, LH and FSH levels along with significant increase in RBCs MDA and NO and drop in RBCs antioxidant enzymes (SOD and GPx). Contrary to the DC group, significant increase in testosterone, LH and FSH levels accompanied with significant decline in RBCs MDA and NO levels in addition to increase in the activity of SOD and GPx were observed in G. kola, fermented probiotics-yoghurt and their combine groups. Among the experimental treatment groups, the combine of G. kola and fermented probiotics-yoghurt showed the most significantly increase in testosterone, LH and FSH levels (1.82, 4.13 and 11.93 ng/ml, respectively), the less MDA and NO levels (9.5 and 1.93 nmol/g Hb, respectively) and the highest activity of SOD and GPx (4.5 and 32.18 U/g Hb, respectively).
Effects of GK, probiotics-yoghurt and their combine on testosterone (A), LH (B), FSH (C), MDA (D), GPx (E), NO (F) and SOD (G). Values are set as mean ± SE (n = 6). At a statistical significance level of P < 0.05, various superscript letters were added to represent group differences
Effects of GK, probiotics-yoghurt and their combine on the lipid profile and dyslipidemia
Regarding the lipid profile and dyslipidemia parameters (Fig. 5), significant elevation in serum cholesterol, TG, LDL, non-HDL-Ch, Ch/HDL ratio and VLDL levels accompanied with significant decrease in HDL level was observed in high-fat diet/low STZ-treated rats versus the NC group. Meanwhile, the administration of G. kola, fermented probiotics-yoghurt and their combine caused a significant (p < 0.05) decline in serum cholesterol, TG, LDL, non-HDL-Ch, Ch/HDL ratio and VLDL levels and significant elevation in HDL level as compared with the DC group. Notably, the combine of G. kola and fermented probiotics-yoghurt significantly showed the most reduction in lipid parameters recording 109.67, 86.23, 46.98 and 38.15 mg/dl for TC, TG, LDL and HDL, respectively and the less atherogenic indices (non-HDL-Ch, VLDL and Ch/HDL ratio) at 71.52, 17.25 mg/dl and 2.88, respectively.
Effects of GK, probiotics-yoghurt and their combine on total cholesterol (A), TG (B), LDL-Ch (C), HDL-Ch (D), Ch/HDL ratio (E), non HDL-Ch (F) and VLDL (G). Values are set as mean ± SE (n = 6). At a statistical significance level of P < 0.05, various superscript letters were added to represent group differences
Effects of GK, probiotics-yoghurt and their combine on the liver and kidney biochemical indicators
Concerning liver an kidney biochemical indicators, Fig. 6 illustrates that the activity of AST, ALT, ALP, LDH and the levels of urea and creatinine were significantly (p < 0.05) higher in DC group than NC group while, albumin and total protein levels were significantly lower in DC group than NC group. Compared to DC group, the administration of G. kola, fermented probiotics-yoghurt and their combine to the diabetic rats led to suppression in the dysfunction of liver and kidney. Despite the converging between the different treatments in their effects on liver and kidney functions, the co-administration of G. kola and fermented probiotics-yoghurt significantly showed the less activity of AST, ALP and LDH and the highest level of total protein.
Effects of GK, probiotics-yoghurt and their combine on AST (A), ALT (B), ALP (C), LDH (D), albumin (E), total protein (F), urea (G) and creatinine (H). Values are set as mean ± SE (n = 6). At a statistical significance level of P < 0.05, various superscript letters were added to represent group differences
Discussion
T2DM is a dangerous chronic disease that poses a threat to the public’s health. It is an endocrine and metabolic disorder primarily defined by high blood glucose levels. Additionally connected to it include obesity, liver damage, insulin resistance, and malfunction of the pancreatic islet cells (Ouyang et al. 2022). In addition to combating diabetes through a number of mechanisms, including glucose utilization in extra-hepatic tissues, a direct diminution in macrophage infiltration, an enhancement in beta cell function, and an increase in the functional activity of glucose transporters, the bioactive phytochemicals of G. kola seeds demonstrated several beneficial effects, including antioxidant and anti-inflammatory (Oyenihi et al. 2022).
Studies using animal models suggest that probiotics may lower blood glucose levels by reducing inflammation and preventing the breakdown of β-pancreatic cells. Consuming probiotics is thought to protect the β-pancreatic cells from oxidative damage by reducing lipid peroxidation and elevating antioxidants like glutathione and peroxide dismutase. Additionally, probiotics have been found to affect insulin resistance and inflammation by boosting natural killer cells and controlling the expression of TNF-α. The increase in gliclazide bioavailability, the blockage or delay of intestinal glucose absorption, and changes to the gut flora are other potential pathways that could have an impact on glucose metabolism (Yanni et al. 2020).
Thus, the freeze-dried aqueous infusion of G. kola seeds, fermented probiotics-yoghurt and both together were evaluated for their potential therapeutic effect of type 2 diabetes and its problems included the dysfunction of male fertility-stimulating hormones. Among the most prevalent secondary metabolites in nature are phenolic compounds. Because they include phenolic groups that donate electrons to other molecules, polyphenols have the ability to act as antioxidants. Few polyphenols are hydrolyzed or absorbed in the stomach because they are resistant to the stomach’s acid. Large amounts of ingested polyphenols interact with the gut microbiota, the indigenous bacteria, in the large intestine. Epidemiological studies demonstrate that a long-term, moderate diet of foods high in polyphenols can halt the progression of chronic illnesses like type 2 diabetes, cardiovascular disease, and neurodegenerative diseases as well as obesity (Bié et al. 2023). Thus, the phenolic profile of the aqueous infusion of G. kola was evaluated in this study and 18 known phenolic compounds in the freeze-dried G. kola infusion were identified. The radical scavenging activity of G. kola is most probably attributed to the phenolic compounds. The greatest way for phenolic compounds to act as antioxidants is via directly scavenging free radicals. A hydrogen atom transport mechanism and an electron transfer mechanism are both involved in this process (Kostić et al. 2023).
Because the high fat diet and STZ treatments are designed to resemble the stages and transitions in the progression of type 2 diabetes (Andonova et al. 2023), the high fat diet and STZ-induced type 2 diabetic rats model was used in this study. The high fat diet significantly increased the body weight gain compared with the NC group. While, diabetic rats were found to have significant weight loss, compared with NC group. Andonova et al. (2023) also found that feeding rats on a diet high in fat, mainly lard both saturated and monounsaturated fatty acids, led to an increase in the weight of rats compared to normal rats, while the weight of those fed on fat decreased after being injected with STZ for ten days as a result of the development of type 2 diabetes. T2DM can display a number of symptoms, including hyperglycemia, hyperlipidaemia, and loss of body weight. High blood glucose levels prevent the absorption of energy sources and eliminated through urination to cause weight reduction (El-Newary et al. 2021). In this study, the different therapies converged greatly in their efficacy on improving diabetes-related weight loss. Data indicate that the administration of G. kola, fermented probiotics-yoghurt or their combine could effectively improve weight loss in T2DM rats. The amalgamation of G. kola and probiotics-yoghurt had the most promising effect on diabetic and its complications, which can be strongly attributed to the richness of seeds in phenolic compounds, as confirmed by the results of the current study since the effective role of phenolic compounds as prebiotics has been indicated, and it has been shown that the combination of phenolic compounds and probiotics may have health-promoting effects (Plamada & Vodnar 2021). The synergistic effect between flavonoids and Bifidobacterium spp was reported by Pan et al. (2023), in 10 healthy Chinese volunteers, an in vitro induced fermentation method was utilized to assess the effect of flavonoids included hesperetin-7-O-glucoside, hesperetin, naringin, naringenin, rutin, isoquercitrin, and quercetin. The results showed that hesperetin-7-O-glucoside and quercetin supplementation greatly increased the abundance of Bifidobacterium spp.
In this study,through the antioxidant, anti-inflammatory, nephroprotective, hepatoprotective, and cardioprotective properties of probiotic strains and the phytochemicals of G. kola seeds, the probiotic yoghurt and garcina infusion represented their ameliorative effect against type 2 diabetes and its complications. FBG level, which represents the blood glucose level in right now, is a crucial reference point in the diagnosis of T2DM. The phytochemicals found in G. kola, mainly flavonoids, saponins, tannins, phenols, glycosides, and alkaloids are involved in the hypoglycemic effect of G. kola infusion. Furthermore, the detected phenolics, namely daidzein and quercetin, exert hypoglycemic effect and suppress inflammation and oxidative stress resulting in attenuating of cardiac, hepatic and renal injury (Laddha & Kulkarni 2021 and Kareem et al. 2022). In this study, the hypoglycemic effect of the probiotic yoghurt strongly attributed to the selected strains, as L. acidophilus could relieve T2DM via control hepatic glucose, lipid metabolism, and gut microbiota in mice (Yan et al. 2019). The combined of L. acidophilus and B. bifidum positively correlated to SCFAs and type 2 diabetes management (Meng et al. 2023).
There is a correlation between blood glucose and serum insulin, reflecting the balance between glucose metabolism, glycogenolysis, and insulin secretion (Li et al. 2022). The results demonstrated that G. kola and probiotic yoghurt decreased the insulin resistant. Such effect might be duo to flavonoids compounds in G. kola (Li et al. 2023) and L. acidophilus along with bifidbacterium in the fermented yoghurt as confirmed by Rezazadeh et al. (2021) who found that probiotic yogurt containing L. acidophilus and B. lactis exhibited antioxidant activity and improved insulin sensitivity in metabolic syndrome-patients.
Inflammatory processes are implicated in diabetes mellitus. However, inflammatory responses could be modulated by pharmacological interventions, to prevent and treat complications of diabetes (Li et al. 2023). The anti-inflammatory effect of garcina infusion might be attributed to flavonoids, namely quercetin, hesperidin, naringenin and rutin detected in garcina seeds and suppress inflammatory mediators such as TNF-α and IL-6, thus controlling NO generation as reported by Li et al. (2023). The anti-inflammatory effect of probiotic yoghurt may correlated to L. acidophilus, Bifidobacterium as these probiotic strains apply a potent anti-inflammatory effect through adjusting toll like receptor 2-mediated NF-κB and MAPK signaling pathways in inflammatory cells (Li et al. 2019).
As diabetes worsens, hyperglycemia induces mitochondrial dysfunction and the production of reactive oxygen species (ROS), which causes oxidative stress in multiple organs, including blood vessels, pancreatic beta cells, liver and kidney (Mohamed & Abdel-Salam 2021). Oxidative stress originates as a result of uncontrollable generation of ROS and decreased the activity of glutathion and SOD as well as mediated lipid peroxidation and protein carbonylation (Singh et al. 2019). This was evident in diabetic rats in this study. The dysfunctional anti-oxidative system was restored by treatment with G kola and probiotic yoghurt. Such effect might be duo to polyphenols and alkaloids in G kola (Kareem et al. 2022) and L. acidophilus along with bifidbacterium in the fermented yoghurt (Rezazadeh et al. 2021).
Pancreatic cells normally release insulin. The intracellular protein tyrosine kinase (PTK) is then activated once insulin connects to their receptors. To activate phosphoinositide 3-kinase (PI3K), activated PTK is able to phosphorylate and activate insulin receptor substrates (IRS). The PI3K signalling cascade is triggered, which increases gonadotropin-releasing hormone (GnRH) production in the hypothalamus. GnRH then drives pituitary secretion of LH and FSH, which in turn triggers the release of sex hormones by the gonads. Accordingly, increased insulin resistance followed by a defect in the production of testosterone, FSH and LH (Liu et al. 2023). Furthermore, in adipose tissue, modified sex hormone binding globulin (SHBG) levels, elevated levels of oxidative stress, and elevated levels of aromatase (CYP19) activity are exist during diabetes. This causes testosterone and androstenedione to be converted into estradiol and estrogen, respectively, which lowers the levels of testosterone in the blood in diabetic males (Liu et al. 2023). The antioxidant and insulin sensitivity stimulating effects of garcinia may be implicated in its effect on improving testosterone, FSH and LH hormone imbalances associated with type 2 diabetes. Additionally, G. kola is one of the plants that have a role against infertility and reproductive dysfunction of males through pretesticular effect by increasing testosterone, FSH and LH levels (Farombi et al. 2013).
The occurrence and progression of diabetic vascular problems are caused by a low-grade inflammatory reaction of the blood vessel wall caused by hyperglycemia. Inflammatory mediators continually activate vascular endothelial cells, and the integrity of the connection between endothelial cells, as well as cell gaps, is damaged as a result of increased vascular permeability and macro-molecular material leakage. When these leaky macro-molecular compounds accumulate in the blood artery wall, they cause tissue edema and accelerate atherosclerosis (Li et al. 2023). Dyslipidemia accompanied by impaired liver and kidney functions were evident in diabetic rats in this study. The disturbance in lipid metabolism and impaired liver and kidney functions were attenuated by treatment with G kola and probiotic yoghurt. The anti-inflammatory effect of G kola phytochemicals and probiotic strains might be contributed to their improvement effect against dyslipidemia and impaired liver and kidney biochemical indicators. The activation of the AMP-activated protein kinase (AMPK) has frequently been linked to the molecular mechanisms behind the beneficial effects of flavonoids. The control of lipid metabolism and adipogenesis depends heavily on the enzyme AMPK. Catabolic activities like glucose absorption and glycolysis are enhanced by AMPK phosphorylation and activation, while anabolic processes like fatty acid synthesis and gluconeogenesis are inhibited (Sandoval et al. 2020). Bifidobacterium, as a probiotic strain, has the potential to reduce blood TC and LDL-C levels in hyperlipidemic patients via gut microbiota modulation. The greater abundance of anti-obesity-related genera and fecal metabolites could be the underlying mechanism (Chu et al. 2023).
Conclusion
The results of the HPLC analysis of freeze-dried aqueous infusion of G. kola demonstrated the presence of several polyphenols such as querectin and hesperetin. Owing to these antioxidant compounds, the freeze-dried aqueous infusion of G. kola exhibited potent radical scavenging activity. By analyzing the results, it was possible to draw the conclusion that the co-administration of the freeze-dried aqueous infusion of G. kola and the fermented probiotics-yoghurt was promising in regulation of glucose, insulin and the male fertility-stimulating hormones in diabetic rats. Additionally, this co-administration resulted in down-regulation of inflammatory markers (CRP, TNF-α, IL-6 and TGF-1β) and oxidative markers (MDA and NO), up-regulation of antioxidant enzymes (SOD and GPx) and reduction of dyslipidemia in type 2 diabetic rats.
Availability of data and materials
The datasets used and/ or analyzed during the current study are available from the corresponding author on reasonable request.
References
Abdel-Salam, A. M., Al Hemaid, W. A., Afifi, A. A., Othman, A. I., Farrag, A. H., & Zeitoun, M. M. (2018). Consolidating probiotic with dandelion, coriander and date palm seeds extracts against mercury neurotoxicity and for maintaining normal testosterone levels in male rats. Toxicology Reports, 5(2018), 1069–1077. https://doi.org/10.1016/j.toxrep.2018.10.013
Abdel-Salam, A. M., El-Ziney, M. G., Zaghloul, A. H., Babiker, A. Y., & Mousa, H. M. (2009). The effectiveness of whey proteins mixed with hot-water extract of Artemisia and Capparis spp. against lead acetate-contamination in rats. Food Agriculture and Environment, 7(3&4), 139–141.
Adedara, I. A., Awogbindin, I. O., Anamelechi, J. P., & Farombi, E. O. (2015). Garcinia kola seed ameliorates renal, hepatic, and testicular oxidative damage in streptozotocin-induced diabetic rats. Pharmaceutical Biology, 53(5), 695–704. https://doi.org/10.3109/13880209.2014.937504
Ajayi, S. A., Ofusori, D. A., Ojo, G. B., Ayoka, O. A., Abayomi, T. A., & Tijani, A. A. (2011). The microstructural effects of aqueous extract of Garcinia kola (Linn) on the hippocampus and cerebellum of malnourished mice. Asian Pacific Journal of Tropical Biomedicine, 1(4), 261–265. https://doi.org/10.1016/s2221-1691(11)60039-7
Al Bataineh, M. T., Künstner, A., Dash, N. R., Alsafar, H. S., Ragab, M., Schmelter, F., Sina, C., Busch, H., & Ibrahim, S. M. (2023). Uncovering the relationship between gut microbial dysbiosis, metabolomics, and dietary intake in type 2 diabetes mellitus and in healthy volunteers: A multi-omics analysis. Scientific Reports, 13(1), 17943. https://doi.org/10.1038/s41598-023-45066-7
Andlib, N., Sajad, M., Kumar, R., & Thakur, S. C. (2023). Abnormalities in sex hormones and sexual dysfunction in males with diabetes mellitus: A mechanistic insight. Acta Histochemica, 125(1), 151974. https://doi.org/10.1016/j.acthis.2022.151974
Andonova, M., Dzhelebov, P., Trifonova, K., Yonkova, P., Kostadinov, N., Nancheva, K., Ivanov, V., Gospodinova, K., Nizamov, N., Tsachev, I., & Chernev, C. (2023). Metabolic markers associated with progression of type 2 diabetes induced by high-fat diet and single low dose streptozotocin in rats. Veterinary Sciences, 10(7), 431. https://doi.org/10.3390/vetsci10070431
Bié, J., Sepodes, B., Fernandes, P. C. B., & Ribeiro, M. H. L. (2023). Polyphenols in health and disease: gut microbiota, bioaccessibility, and bioavailability. Compounds, 3(1), 40–72. https://doi.org/10.3390/compounds3010005
Burstein, M., Scholnick, H. R., & Morfin, R. (1970). Rapid method for the isolation of lipoproteins from human serum by precipitation with polyanions. Journal of Lipid Research, 11(6), 583–595. https://doi.org/10.1016/s0022-2275(20)42943-8
Cetkovic-Cvrlje, M., Rogan, S., & Barbaro, E. (2022). Garcinia kola treatment exhibits immunomodulatory properties while not affecting type 1 diabetes development in an experimental mouse model. International Journal of Immunopathology and Pharmacology, 36, 20587384211069830. https://doi.org/10.1177/20587384211069831
Cho, N. H., Shaw, J. E., Karuranga, S., Huang, Y., Fernandes, J. D. R., Ohlrogge, A. W., & Malanda, B. (2018). IDF Diabetes Atlas: Global estimates of diabetes prevalence for 2017 and projections for 2045. Diabetes Research and Clinical Practice, 138, 271–281. https://doi.org/10.1016/j.diabres.2018.02.023
Chu, C., Jiang, J., Yu, L., Li, Y., Zhang, S., Zhou, W., Wang, Q.,Zhao, J., Zhai, Q., Tian, F., & Chen, W. (2023). Bifidobacterium longum CCFM1077 attenuates hyperlipidemia by modulating the gut microbiota composition and fecal metabolites: a randomized, double-blind, placebo-controlled clinical trial. Engineering. https://doi.org/10.1016/j.eng.2023.04.010. In Press.
Doumas, B. T., Watson, W. A., & Biggs, H. G. (1997). Albumin standards and the measurement of serum albumin with bromcresol green. Clinica Chimica Acta, 258(1), 21–30. https://doi.org/10.1016/s0009-8981(96)06447-9
El-Newary, S. A., Afifi, S. M., Aly, M. S., Ahmed, R. F., El Gendy, A. N., Abd-ElGawad, A. M., Farag, M. A., Elgamal, A. M., & Elshamy, A. I. (2021). Chemical profile of Launaea nudicaulis ethanolic extract and its antidiabetic effect in streptozotocin-induced rats. Molecules, 26(4), 1000. https://doi.org/10.3390/molecules26041000
Farahna, M., Etet, P. F. S., Osman, S. Y., Yurt, K. K., Amir, N., Vecchio, L., Aydin, I., Aldebasi, Y. H., Sheikh, A., Chijuka, J. C., Kaplan, S., & Adem, A. (2017). Garcinia kola aqueous suspension prevents cerebellar neurodegeneration in long-term diabetic rat–a type 1 diabetes mellitus model. Journal of Ethnopharmacology, 195, 159–165. https://doi.org/10.1016/j.jep.2016.11.001
Farombi, E. O., Adedara, I. A., Oyenihi, A. B., Ekakitie, E., & Kehinde, S. (2013). Hepatic, testicular and spermatozoa antioxidant status in rats chronically treated with Garcinia kola seed. Journal of Ethnopharmacology, 146(2), 536–542. https://doi.org/10.1016/j.jep.2013.01.018
Fawcett, J. K., & Scott, J. E. (1960). A rapid and precise method for the determination of urea. Journal of Clinical Pathology, 13(2), 156–159. https://doi.org/10.1136/jcp.13.2.156
Gomez, S., Anjali, C., Kuruvila, B., Maneesha, P. K., & Joseph, M. (2023). Phytochemical constitution and antioxidant activity of functional herbal drink from Indian gooseberry (Emblica officinalis Gaertn.) fruits containing spices and condiments. Food Production, Processing and Nutrition, 5(1), 1–13. https://doi.org/10.1186/s43014-022-00127-8
Idris, A. E., Etet, P. F. S., Saeed, A. A., Farahna, M., Satti, G. M., AlShammari, S. Z., & Hamza, M. A. (2020). Evaluation of metabolic, antioxidant and anti-inflammatory effects of Garcinia kola on diabetic rats. Saudi Journal of Biological Science, 27(12), 3641–3646. https://doi.org/10.1016/j.sjbs.2020.08.006
Kareem, A., Josiah, S. S., Saliu, I. O., Olayeriju, O. S., Akinmoladun, A. C., & Akindahunsi, A. A. (2022). Ameliorative influence of Garcinia kola seed extracts against multiple organ toxicity in monosodium glutamate-administered Wistar rats. Comparative Clinical Pathology, 31(6), 987–1004. https://doi.org/10.1007/s00580-022-03406-5
Kostić, K., Brborić, J., Delogu, G., Simić, M. R., Samardžić, S., Maksimović, Z., Dettori, M. A., Fabbri, D., Kotur-Stevuljević, J., & Saso, L. (2023). Antioxidant activity of natural phenols and derived hydroxylated biphenyls. Molecules, 28(6), 2646. https://doi.org/10.3390/molecules28062646
Laddha, A. P., & Kulkarni, Y. A. (2021). Daidzein mitigates myocardial injury in streptozotocin-induced diabetes in rats. Life Science, 284, 119664. https://doi.org/10.1016/j.lfs.2021.119664
Larsen, K. (1972). Creatinine assay by a reaction-kinetic principle. Clinica Chimica Acta, 41(C), 209–217. https://doi.org/10.1016/0009-8981(72)90513-X
Li, L., Zhang, Y., Luo, Y., Meng, X., Pan, G., Zhang, H., Li, Y., & Zhang, B. (2023). The molecular basis of the anti-inflammatory property of astragaloside IV for the treatment of diabetes and its complications. Drug Design, Development and Therapy, 17, 771–790. https://doi.org/10.2147/DDDT.S399423
Li, M., Chi, X., Wang, Y., Setrerrahmane, S., Xie, W., & Xu, H. (2022). Trends in insulin resistance: Insights into mechanisms and therapeutic strategy. Signal Transduction and Targeted Therapy, 7(1), 216. https://doi.org/10.1038/s41392-022-01073-0
Li, S. C., Hsu, W. F., Chang, J. S., & Shih, C. K. (2019). Combination of Lactobacillus acidophilus and Bifidobacterium animalis subsp. lactis shows a stronger anti-inflammatory effect than individual strains in HT-29 cells. Nutrients, 11(5), 969. https://doi.org/10.3390/nu11050969
Liu, J., Liu, Z., Sun, W., Luo, L., An, X., Yu, D., & Wang, W. (2023). Role of sex hormones in diabetic nephropathy. Frontiers in Endocrinology, 14, 1135530. https://doi.org/10.3389/fendo.2023.1135530
Luo, J., Jiang, L., Gao, B., Chai, Y., & Bao, Y. (2023). Comprehensive in silico analysis of the probiotics, and preparation of compound probiotics-Polygonatum sibiricum saponin with hypoglycemic properties. Food Chemistry, 404, 134569. https://doi.org/10.1016/j.foodchem.2022.134569
Mansour, A., Mousa, M., Abdelmannan, D., Tay, G., Hassoun, A., & Alsafar, H. (2023). Microvascular and macrovascular complications of type 2 diabetes mellitus: Exome wide association analyses. Frontiers in Endocrinology, 14, 1143067. https://doi.org/10.3389/fendo.2023.1143067
Megraw, R., Dunn, D., & Biggs, H. (1979). Manual and continuous flow colorimetry of triglycerols by a fully enzymatic method. Clinical Chemistry, 25(2), 273–278. https://doi.org/10.1093/clinchem/25.2.273
Meng, F., Zhang, F., Meng, M., Chen, Q., Yang, Y., Wang, W., Xie, H., Li, X., Gu, W., & Yu, J. (2023). Effects of the synbiotic composed of mangiferin and Lactobacillus reuteri 1–12 on type 2 diabetes mellitus rats. Frontiers in Microbiology, 14, 1158652. https://doi.org/10.3389/fmicb.2023.1158652
Mohamed, R. S., & Abdel-Salam, A. M. (2021). Efficiency of a formulated condiment (duqqa) in mitigation of diabetes and its complications induced by streptozotocin-nicotinamide in rats. Journal of Herbmed Pharmacology, 10(2), 218–225. https://doi.org/10.34172/jhp.2021.24
Montgomery, H. A. C. D. J., & Dymock, J. F. (1961). Determination of nitrite in water. The Analyst, 86(102), 414.
Nguyen, T. T. K., Laosinwattana, C., Teerarak, M., & Pilasombut, K. (2017). Potential antioxidant and lipid peroxidation inhibition of Phyllanthus acidus leaf extract in minced pork. Asian-Australas Journal of Animal Science, 30(9), 1323–1331. https://doi.org/10.5713/ajas.17.0036
Nishikimi, M., Rao, N. A., & Yagi, K. (1972). The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochemical and Biophysical Research Communications, 46(2), 849–854. https://doi.org/10.1016/s0006-291x(72)80218-3
Novelli, E. L. B., Diniz, Y. S., Galhardi, C. M., Ebaid, G. M. X., Rodrigues, H. G., Mani, F., Fernandes, A. A. H., Cicogna, A. C., & Novelli Filho, J. L. V. B. (2007). Anthropometrical parameters and markers of obesity in rats. Laboratory Animals, 41(1), 111–119. https://doi.org/10.1258/002367707779399518
Oduwole, O. O., Huhtaniemi, I. T., & Misrahi, M. (2021). The roles of luteinizing hormone, follicle-stimulating hormone and testosterone in spermatogenesis and folliculogenesis revisited. International Journal of Molecular Science, 22(23), 12735. https://doi.org/10.3390/ijms222312735
Ogunkunle, A. T., Oyelakin, T. M., Enitan, A. O., & Oyewole, F. E. (2014). A quantitative documentation of the composition of two powdered herbal formulations (antimalarial and haematinic) using ethnomedicinal information from ogbomoso, Nigeria. Evidence-Based Complementary and Alternative Medicine, 2014, 751291. https://doi.org/10.1155/2014/751291
Ohkawa, H., Ohishi, N., & Yagi, K. (1979). Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry, 95(2), 351–358. https://doi.org/10.1016/0003-2697(79)90738-3
Ouyang, Y., Liu, D., Zhang, L., Li, X., Chen, X., & Zhao, C. (2022). Green alga enteromorpha prolifera oligosaccharide ameliorates ageing and hyperglycemia through gutbrain axis in age-matched diabetic mice. Molecular Nutrition and Food Research, 66(4), 210056. https://doi.org/10.1002/mnfr.202270011e2100564
Oyenihi, O. R., Cerf, M. E., Matsabisa, M. G., Brooks, N. L., & Oguntibeju, O. O. (2022). Effect of kolaviron on islet dynamics in diabetic rats. Saudi Journal of Biological Science, 29(1), 324–330. https://doi.org/10.1016/j.sjbs.2021.08.095
Paglia, D. E., & Valentine, W. N. (1967). Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. Journal of Laboratory and Clinical Medicine, 70(1), 158–169.
Pan, L., Ye, H., Pi, X., Liu, W., Wang, Z., Zhang, Y., & Zheng, J. (2023). Effects of several flavonoids on human gut microbiota and its metabolism by in vitro simulated fermentation. Frontiers in Microbiology, 2023(14), 1092729.
Plamada, D., & Vodnar, D. C. (2021). Polyphenols—gut microbiota interrelationship: A transition to a new generation of prebiotics. Nutrients, 14(1), 137. https://doi.org/10.3390/nu14010137
Prior, R. L., Wu, X., & Schaich, K. (2005). Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements. Journal of Agriculture and Food Chemistry, 53(10), 4290–4302. https://doi.org/10.1021/jf0502698
Qian, C., Zhu, C., Yu, W., Jiang, X., & Zhang, F. (2015). High-fat diet/low-dose streptozotocin-induced type 2 diabetes in rats impacts osteogenesis and Wnt signaling in bone marrow stromal cells. PLoS One, 10(8), e0136390. https://doi.org/10.1371/journal.pone.0136390
Reeves, P. G., Nielsen, F. H., & Fahey, G. C. (1993). AIN-93 purified diets for laboratory rodents: Final report of the American institute of nutrition Ad hoc writing committee on the reformulation of the AIN-76A rodent diet. Journal of Nutrition, 123, 1939–1951. https://doi.org/10.1093/jn/123.11.1939
Reitman, S., & Frankel, S. (1957). A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. American Journal of Pathology, 28(1), 56–63. https://doi.org/10.1093/ajcp/28.1.56
Rezazadeh, L., Alipour, B., Jafarabadi, M. A., Behrooz, M., & Gargari, B. P. (2021). Daily consumption effects of probiotic yogurt containing Lactobacillus acidophilus La5 and Bifidobacterium lactis Bb12 on oxidative stress in metabolic syndrome patients. Clinical Nutrition, 41, 136–142. https://doi.org/10.1016/j.clnesp.2020.12.003
Rheinhold, J., & Seligron, D. (1953). Total protein, albumin and globulin in standard methods of clinical chemistry (pp. 88). J Academic Press, Inc.
Sandoval, V., Sanz-Lamora, H., Arias, G., Marrero, P. F., Haro, D., & Relat, J. (2020). Metabolic impact of flavonoids consumption in obesity: from central to peripheral. Nutrients, 12(8), 2393. https://doi.org/10.3390/nu12082393
Schriewer, H., Kohnert, U., & Assmann, G. (1984). Determination of LDL cholesterol and LDL apolipoprotein B following precipitation of VLDL in blood serum with phosphotungstic acid/MgCl2. Clinical Chemistry and Laboratory Medicine, 22(1), 35–40. https://doi.org/10.1515/cclm.1984.22.1.35
Singh, A., Kukreti, R., Saso, L., & Kukreti, S. (2019). Oxidative stress: A key modulator in neurodegenerative diseases. Molecules, 24(8), 1583. https://doi.org/10.3390/molecules24081583
Singleton, V. L., Orthofer, R., & Lamuela-Raventos, R. M. (1999). Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods in Enzymology, 299, 152–178. https://doi.org/10.1016/s0076-6879(99)99017-1
Srinivasan, K., Viswanad, B., Asrat, L., Kaul, C. L., & Ramarao, P. (2005). Combination of high-fat diet-fed and low-dose streptozotocin-treated rat: A model for type 2 diabetes and pharmacological screening. Pharmacological Research, 52(4), 313–320. https://doi.org/10.1016/j.phrs.2005.05.004
Tamime, A. Y., & Robinson, R. K. (1999). Yoghurt: Science and technology. Wood head Publishers.
Trinder, P. (1969). Determination of blood glucose using an oxidase-peroxidase system with a non-carcinogenic chromogen. Journal of Clinical Pathology, 22(2), 158–161. https://doi.org/10.1136/jcp.22.2.158
Van Cauwenberghe, J., De Block, C., Vanderschueren, D., & Antonio, L. (2022). Effects of treatment for diabetes mellitus on testosterone concentrations: A systematic review. Andrology, 11(2), 225–233. https://doi.org/10.1111/andr.13318
Wang, H., Fu, Y., Zhao, Q., Liu, Z., Wang, C., Xue, Y., & Shen, Q. (2023). Effects of heat-treated starch and protein from foxtail millet (Setaria italica) on type 2 diabetic mice. Food Chemistry, 404, 134735. https://doi.org/10.1016/j.foodchem.2022.134735
Watson, D. (1960). A simple method for the determination of serum cholesterol. Clinica Chimica Acta, 5(5), 637–642. https://doi.org/10.1016/0009-8981(60)90004-8
Yan, F., Li, N., Shi, J., Li, H., Yue, Y., Jiao, W., Wang, N., Song, Y., Huo, G., & Li, B. (2019). Lactobacillus acidophilus alleviates type 2 diabetes by regulating hepatic glucose, lipid metabolism and gut microbiota in mice. Food and Function, 10(9), 5804–5815. https://doi.org/10.1039/c9fo01062a
Yanni, A. E., Kartsioti, K., & Karathanos, V. T. (2020). The role of yoghurt consumption in the management of type II diabetes. Food and Function, 11(12), 10306–10316. https://doi.org/10.1039/d0fo02297g
Zhu, Z., Lin, Z., Jiang, H., Jiang, Y., Zhao, M., & Liu, X. (2017). Hypolipidemic effect of Youcha in hyperlipidemia rats induced by high-fat diet. Food and Function, 8(4), 1680–1687. https://doi.org/10.1039/c7fo00089h
Zimmerman, H. J., & Weinstein, H. G. (1956). Lactic dehydrogenase activity in human serum. Journal of Laboratory and Clinical Medicine, 48(4), 607–616.
Acknowledgements
The authors are thankful for the technical support by the National Research Centre, Egypt.
Funding
None.
Author information
Authors and Affiliations
Contributions
R.S.M. Conceptualization, animal experiment, analysis, sampling, methodology, writing- original draft preparation. K.F. Conceptualization, animal experiment, analysis, sampling, methodology, writing- original draft preparation. A.H.Z. Conceptualization, sampling, methodology, writing- original draft preparation. A.M.A. Conceptualization, sampling, methodology, writing- original draft preparation.
Corresponding author
Ethics declarations
Ethics approval and consent to participate
The study has been approved by the Medical Research Ethics Committee, National Research Centre, Cairo, Egypt (No.24911122022) and all the procedures on animals were carried out according to guidelines and recommendations of the above committee.
Consent for publication
Not applicable.
Competing interests
The authors declare that they have no competing interests.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
About this article
Cite this article
Mohamed, R.S., Fouda, K., Zaghloul, A.H. et al. Potential antidiabetic activity of probiotic and Garcinia kola-yoghurt and its role in regulation of male fertility-stimulating hormones in high-fat diet/low dose streptozotocin-treated rats. Food Prod Process and Nutr 6, 55 (2024). https://doi.org/10.1186/s43014-024-00229-5
Received:
Accepted:
Published:
DOI: https://doi.org/10.1186/s43014-024-00229-5