Berberine reinforces Sertoli cells niche and accelerates spermatogonial stem cells renewal in experimentally-induced varicocele condition in rats

Abstract Background Varicocele is present in 10–20% of the male infertile population. Purpose: Present study was done to demonstrate the reinforcing effect of berberine (BBR), as an antioxidant and anti-inflammatory agent, on Sertoli cells-related niche and spermatogonial stem cells (SSCs) self-renewal in experimentally-induced VCL condition. Study design 50 mature male Wistar rats were divided into control, control-sham, non-treated VCL-induced, 50 mg kg−1 and 100 mg kg−1 BBR-treated VCL-induced groups. Methods The Leydig and Sertoli cells distribution and Leydig cells steroidogenic activity, expression of glial cell line-derived neurotrophic factor (GDNF), proto-oncogene Rearranged during Transfection (c-RET) receptor, Ets variant gene 5 (Etv5) and B-cell chronic lymphocytic leukemia (CLL)/lymphoma 6, member B (Bcl-6b) at mRNA and protein levels were analyzed. The mRNA integrity and DNA fragmentation were assessed. Finally, the serum levels of testosterone, inhibin B and testicular total antioxidant capacity, total thiol molecules, catalase, and malondialdehyde were evaluated. Results Observations revealed that, the BBR significantly enhanced VCL-reduced Leydig and Sertoli cells population, maintained Leydig–Sertoli cells network, enhanced GDNF, c-RET Etv5 and Bcl6b expression, up-regulated testicular antioxidant and endocrine status. Conclusion The BBR by boosting Leydig–Sertoli cells network up-regulates the GDNF, Etv5 and Bcl-6b expression/synthesis in SSCs, which in turn improves SSCs self-renewal activities. Thus, the BBR could be considered as an appropriate agent for antioxidant therapy of VCLs. However, more studies with bigger sample number and focus on BBR-induced effects on other genes involving in the self-renewal process are needed to have more deterministic results. Keywords Varicocele Berberine Spermatogonial stem cells Self-renewa

One-carbon cycle support rescues sperm damage in experimentally induced varicocoele in rats

Objectives: To investigate whether micronutrients in support of the one-carbon cycle and glutathione synthesis are effective in improving sperm damage after surgical varicocoele induction in rats and whether any effect is achieved without a rebound reductive stress as seen with oral antioxidants. Materials and Methods: Surgical varicocoele was induced in adult male Wistar rats and resulted in significant damage to the testis and sperm cells measured at 2 and 4 months after surgery. At 2 months after surgery, rats received a 2-month oral supplementation in support of the one-carbon cycle containing B vitamins (B2, B3, B6, folic acid and B12), N-acetyl-cysteine, zinc, small amounts of vitamin E, and a natural source of betalains and quercetine (Condensyl ® ; Parthenogen SAGL, Lugano, Switzerland and Nurilia SARL, Lyon, France). Results: One-carbon cycle supplementation, compared to untreated controls, significantly improved the morphometric characteristics of testis (P < 0.05), sperm concentration, motility and abnormal morphology (P < 0.001), sperm chromatin condensation (aniline blue staining, P < 0.05), sperm DNA damage (acridine orange staining, P < 0.05) and sperm lipid peroxidation (BODIPY C11, P < 0.001). The improvement in both nuclear condensation and DNA damage and the lack of excessive inhibition of lipid peroxidation confirmed that no reductive stress had occurred. Conclusions: Micronutrients in support of the one-carbon cycle are effective in the treatment of surgically induced varicocoele in rats, probably by activating natural antioxidant defences and epigenetics. These results support the idea that essential micronutrients including B vitamins may also have a positive influence in clinical varicocoele, which should be tested in prospective clinical trials. © 2018 The Authors BJU International © 2018 BJU International Published by John Wiley & Sons Lt

The protective effect of coenzyme Q10 and berberine on sperm parameters, with and without varicocelectomy in rats with surgically induced varicoceles

The current study aimed to investigate the protective effects of coenzyme Q10 (Co Q10) and berberine (BB) with and without varicocelectomy on sperm parameters in postoperative varicocele rats. For the current purpose, a total of 60 mature male Wistar rats were randomly divided into control (n = 6 rats), control-sham (n = 6 rats), and experimental (n = 6 rats) groups. The animals in the experimental groups were undergone experimental varicocele, and simple laparotomy was performed in control-sham group. The experimental group was subdivided into the following groups 60 days after varicocele (VCL) induction: non-treated VCL-induced rats (n = 6 rats), VCL-induced rats administered 100 mg (kg per day) BB (n = 6 rats), VCL-induced rats administered Co Q10 75 mg (kg per day) (n = 6 rats), VCL-induced rats administered 100 mg (kg per day) BB + Co Q10 75 mg (kg per day) (n = 6 rats), varicocelectomy rats (n = 6 rats), varicocelectomy rats administered 100 mg (kg per day) BB (n = 6 rats), varicocelectomy rats administered Co Q10 75 mg (kg per day) (n = 6 rats), varicocelectomy rats administered 100 mg (kg per day) BB + Co Q10 75 mg (kg per day) (n = 6 rats). Following 60 days, the animals were euthanized and sperm parameters were evaluated. Non-treated VCL-induced animals indicated a significant (P < 0.05) decrease in sperm parameters and a significant (P < 0.05) increase in sperm DNA damage compared to control and control-sham groups. Insignificant changes were found between control and control-sham groups. Meanwhile, each treatment group showed a remarkable (P < 0.05) increase in sperm parameters as well as a significant (P < 0.05) decrease in sperm DNA damage. Based on current results, BB and Co Q10 alone and/or together could improve sperm parameters and reduce sperm DNA damage in varicocele-induced rats compared to control and control-sham groups. Varicocelectomy alone will improve sperm parameters, but this recovery will be greater when combined with Co Q10 and BB. © 2018, Springer-Verlag London Ltd., part of Springer Nature

Evaluation of the serum sex hormones levels and alkaline phosphatase activity in rats� testis after administering of berberine in experimental varicocele

Current study was aimed to investigate the protective effect of berberine (BB) on the serum gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B (INHB), testosterone (T) and alkaline phosphatase (Alk-p) activity in the testis of experimental varicocele-induced animals. For the current objective, 30 mature-male Wistar rats were randomly divided into control (n = 6 rats), control-sham (n = 6 rats) and experimental groups (n = 18 rats). The animals in the experimental groups were undergone experimental varicocele and simple laparotomy was conducted in control-sham group. 60 days after varicocele (VCL) induction the experimental group subdivided into: non-treated VCL-induced and 50 mg/kg and 100 mg/kg BB-treated groups (intra-peritoneally). Following 60 days, the animals were euthanized and serum levels of testosterone and testicular activity of alkaline phosphatase were measured. Non-treated VCL-induced animals indicated a significant (P < 0.05) reduction in serum levels of T and INHB and a remarkable (P < 0.05) increase in GnRH, FSH, LH and Alk-p activity compared to control and control-sham groups. Insignificant changes were found between control and control-sham groups. Meanwhile, each BB administered group showed a remarkable (P < 0.05) increase in serum levels of T and INHB and a significant (P < 0.05) decrease in GnRH, FSH, LH and alkaline phosphatase activity in testis tissue. According to the current findings, BB by increasing serum levels of testosterone and INHB increases the testicular endocrine capacity and protects Leydig cell against inflammatory and oxidant injury of varicocele. In addition, BB by inhibiting GnRH, FSH, LH and alkaline phosphatase activity, regulate the levels of serum sex hormones in experimental varicocele and reduces varicocele-induced inflammatory reactions. © 2019, Institute of Korean Medicine, Kyung Hee University

Neuroprotective Effects of Oxytocin Hormone after an Experimental Stroke Model and the Possible Role of Calpain-1

Background Different mechanisms will be activated during ischemic stroke. Calpain proteases play a pivotal role in neuronal death after ischemia damage through apoptosis. Anti-apoptotic activities of the oxytocin (OT) in different ischemic tissues were reported in previous studies. Recently, a limited number of studies have noted the protective effects of OT in the brain. In the present study, the neuroprotective potential of OT in an animal model of transient middle cerebral artery occlusion (tMCAO) and the possible role of calpain-1 in the penumbra region were assessed. Methods Adult male Wistar rats underwent 1 hour of tMCAO and were treated with nasal administration of OT. After 24 hours of reperfusion, infarct size was evaluated by triphenyltetrazolium chloride. Immunohistochemical staining and Western blotting were used to examine the expression of calpain-1. Nissl staining was performed for brain tissue morphology evaluation. Results OT reduced the infarct volume of the cerebral cortex and striatum compared with the ischemia control group significantly (P < .05). Calpain-1 overexpression, which was caused by ischemia, decreased after OT administration (P < .05). The number of pyknotic nuclei in neurons increased dramatically in the ischemic area and OT attenuated the apoptosis of neurons in the penumbra region (P < .01). Conclusion We provided evidence for the neuroprotective role of OT after tMCAO through calpain-1 attenuation. Key Words Stroke tMCAO calpain-1 oxytoci

Berberine ameliorates experimental varicocele-induced damages at testis and sperm levels; evidences for oxidative stress and inflammation

The present study was performed to show the ameliorative effect of berberine (BBR), as an antioxidant and anti-inflammatory agent, against experimental varicocele (VCL)-induced molecular and histological damages. For this purpose, 50 mature Wistar rats were divided into control, control-sham, VCL-sole, 50 mg/kg and 100 mg/kg BBR-treated VCL-induced groups. The tissue levels of interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), nitric oxide (NO), total antioxidant capacity (TAC), malondialdehyde (MDA), superoxide dismutase (SOD) and gluthatione peroxidase (GSH-px) as well as the mRNA levels of testicular CuZn SOD, MnSOD, EC-SOD and GSH-px were evaluated. The serum concentration of testosterone and germ cells mRNA damage were analysed. Finally, the sperm viability, motility, DNA integrity and chromatin condensation were analysed. Observations revealed that, the BBR significantly downregulated VCL-increased IL-6, TNF-α and NO levels, upregulated the CuZn SOD, MnSOD, EC-SOD and GSH-px mRNA level, decreased testicular MDA content, enhanced serum testosterone level and ameliorated testicular TAC, SOD and GSH-px levels. The animals in BBR-treated groups exhibited diminished mRNA damage versus non-treated VCL-induced group. The BBR has significantly (p < 0.05) improved sperm parameters. In conclusion, the BBR by promoting testicular antioxidant potential and by downregulating inflammatory reactions fairly promotes spermatogenesis and upregulates the sperm quality. © 2018 Blackwell Verlag Gmb

Effect of diet contains sesame seed on epididymal histology of adult rat

Sesamin is a major lignan constituent of sesame seed and considered as a key factor in a number of beneficial effects on human health. Sesame leaves intake improve and increase epididymal spermatocytes reserve in adult male sprague dawley rat. The present study was aimed to investigate the effects of contain sesame seed on epididymis histologhical structure. Thirty adult male rats were divided into two groups of 15 rats each. The regimen group received diet containing 30 sesame seed, while the control group received diet along 12 weeks. The right epididymis were removed and minced into several pieces on a specimen bottle containing normal saline for some few minutes to allow the sperms to become motile and swim out and sperm parameters were analyzed. Left epididymis were divided into three sections and fixed into bouin,s solution for further investigation. Serum FSH and LH concentration were estimated by ELISA technique, Testosterone concentration was done by using Chemo-Luminance method. The body weight gained during the treatment period did not differ significantly among groups. The mean epididymal sperm motility and count of the experimental group was significantly higher than control group. LH levels significantly increased in experimental group compared to controls but significant changes in FSH and testosterone levels were not observed in both groups. The results obtained showed that the mean epididymal diameter of the tubular, lumen and epithelium in three parts were not significant in two groups. It can be concluded that sesame seed improves sperm parameters (motility and count) and also can increase LH. But diet contains sesame seed did not affect on epididymal tissue and body weight

The effect of vitamin E & vitamin B on sperm function in rat varicocele model

We assessed the effect of vitamin E and vitamin B (Vit E & Vit B) and their combination on sperm functional parameters in the rat varicocele model. Male rats (n = 120) were divided into control (n = 30), sham (n = 30) and varicocele induction (n = 60) groups. After 2 months, 10 rats from each group were sacrificed to verify varicocele model. This part of results showed that sperm parameters, DNA damage, lipid peroxidation and residual histone were adversely effected in the varicocele group. From the 50 remaining rats in varicocele group, 10 rats received Vit B complex (6, 9.6, 30.4, 9.6 and 0.006 mg/kg for B6, B2, B9, B1 and B12 respectively), 10 rats received Vit E (40 mg/kg), 10 rats received Vit B & E, 10 rats only received water and 10 rats were only received sesame oil as a solvent for Vit E, for 2 months. From 40 remaining rats in control and sham groups, 20 rats only received water and other 20 rats only received sesame oil for 2 months. Then, all the aforementioned parameters were assessed. These results showed that Vit B antioxidant was more efficient in improvement of sperm parameters, chromatin integrity and lipid peroxidation in varicocelized rats compared with Vit E. © 2019 Blackwell Verlag Gmb