Effect of dilute Ethanol Intake on DEHP (di (2-ethylhexyl) phthalate)-induced Testicular Atrophy

Background: Di(2-ethylhexyl) phthalate (DEHP) is the most commonly used as a plasticizer for polyvinyl chloride (PVC), but recently, concern has arisen over the DEHP which may act as a reproductive toxicant to humans. On the other hand, ethanol is the most common supplement of beverages and foods, and so many persons ingest a large quantity of ethanol in daily life. However, interactions between ethanol and DEHP toxicity are not well known. Method: To investigate the effect of dilute ethanol ingestion on the DEHP induced testicular atrophy, rats were received a 1% (w/w) DEHP diet and 2.5 or 5% (v/v) ethanol water for 7 days. Result: The rats treated with DEHP-diet alone for 7 days were observed significant testicular weight loss. On the other hand, testicular weight loss was significantly suppressed in rats treated with DEHP diet and ethanol water. A significant negative correlation between relative testicular weight (as a percentage of body weight) and testicular MEHP concentration was found among rats treated with DEHP-free diet (Control) and DEHP diet alone. Most of the data plots for the DEHP diet plus ethanol water group were scattered above the regression line. Conclusion: These results suggest that dilute ethanol may be effective in preventing DEHP testicular atrophy. However, the mechanism of prevention is unknown and further research is needed.

ZDHHC19 localizes to the cell membrane of spermatids and is involved in spermatogenesis

Sperm is the ultimate executor of male reproductive function. Normal morphology, quantity, and motility of sperm ensure the normal reproductive process. Palmitoylation is a posttranslational modification mediated by palmitoyltransferases whereby palmitoyl is added to proteins. Seven palmitoyltransferases have been identified in Saccharomyces cerevisiae and 23 in humans (including ZDHHC1–9 and ZDHHC11–24), with corresponding homologs in mice. We identified two testis-specific palmitoyltransferases ZDHHC11 and ZDHHC19 in mice. The Zdhhc11 and Zdhhc19-knockout mouse models were constructed, and it was found that the Zdhhc11 knockout males were fertile, while Zdhhc19 knockout males were sterile. ZDHHC19 is located in the cell membrane of step 4–9 spermatids in the mouse testis, and phenotypic analysis showed that the testicular weight ratio in the Zdhhc19−/− mice decreased along with the number and motility of the sperm decreased, while sperm abnormalities increased, mainly due to the “folded” abnormal sperm caused by sperm membrane fusion, suggesting the involvement of ZDHHC19 in maintaining membrane stability in the male reproductive system. In addition, Zdhhc19−/− mice showed abnormal sperm morphologies and apoptosis during spermatogenesis, suggesting that spermatogenesis in the Zdhhc19−/− mice was abnormal. These results indicate that ZDHHC19 promotes membrane stability in male germ cells. Summary sentence: ZDHHC19 is located in the cell membrane of Step4–9 spermatids in mouse testis; Zdhhc19 knockout mice showed male infertility, abnormal spermatogenesis, sperm morphology and motility.

Effect Of Suvarna Bhasma (Gold Calx) On Reproductive System Of Male Albino Rats

Suvarna Bhasma” (gold calx) (SB) prepared as per Ayurveda, a practicing medical system, is prescribed as a medicine in several ailments including conditions like oligozoospermia and asthenoszoospermia in male infertility. SB is prepared from pure metal gold by one of the processing methods detailed in Ayurveda classics. Gold along with other elements was reported in normal human seminal plasma. . In human pathological semen samples, the level of gold was less than normal. Gold may have a role in physiological activity of sperm as shown in case of other elements. The present study supports the hypothesis that the presence of gold is essential for sperm motility. Here we studied the effect of SB on male reproductive organs and semen from epididymis of rat. Present study was performed using Charles foster strain albino rats. Rats were divided into two groups of ten each namely, Group 1 (control) and Group 2 (study). Animals in study group were orally given a fixed amount of SB for 15 days. At the end of the study treated animals showed increase in body weight (<0.05) and testicular weight. In treated animals total sperm count (<0.05) and percentage of sperm motility were increased in epididymal fluid. Histological study showed increase in interstitial area of testes (<0.001), proliferation and branching of the epithelial layer of seminal vesicle. In the Study (gold treated) Group, the increased gold level in genital system may be responsible for the increase in sperm production and motility.

Effect of Methanol Extract, Methanol Fraction, and N-Hexane Fraction of Parijoto Fruit (Medinilla speciosa) on Seminal Parameters and Testicular Weight of Male Sprague Dawley Rats

Infertility occurs in 2 million couples or 17% of couples who are married more than 2 years but are not pregnant or have signs of pregnancy. In couples who do not have children, 50% of male infertility factors are caused by abnormalities in semen. Empirically, parijoto fruits (Medinilla speciosa Blume) are used by the people of Kudus, Central Java, Indonesia to increase fertility. The purpose of this research was to find out the effect of methanol extract, methanol fraction, and n-hexane fraction of parijoto fruit (Medinilla speciosa) on seminal parameters and testicular weight of male Sprague Dawley rats. This research used 36 two-month-old Sprague Dawley rats with 200-300 gram of body weight which were divided into 6 groups. Group 1 (normal); groups 2, 3, and 4, used parijoto fruit methanol extract at 100mg/kgBW, 250mg/kgBW, and 500mg/kgBW doses respectively; groups 5 and 6 used methanol fraction and n-hexane fraction of parijoto fruit at 500 mg/kgBW dose respectively for 14 days. Rats were dissected and had examinations on sperm motility, morphology, abd testicular weight. The data were analyzed using Kruskal Wallis and Mann Whitney. The results found that groups 1, 2, 3, 4, 5 and 6 had mean spermatozoa motility (%) that were 56.5±2.43, 72.5±6.89, 77.6±12.99, 83.3±7.53, 84.7±3.98, and 74.2±11.58, mean spermatozoa morphology (%) that were 95.5±3.67, 95±2.76, 92.6±5.13, 96.5±3.27, 94.3±4.37, and 93.2±6.11, and mean testicular weight (gram) that were 1.08±0.10, 0.90±0.14, 0.98±0.10, 1.18±0.21, 1.28±0.43, and 1.02±0.13. There were significant differences between the normal group and all treatments (P <0.05) on spermatozoa motility. There was no significant difference on spermatozoa morphology and testicular weight. Based on the results, it can be concluded that methanol extract, methanol fraction, and n-hexane fraction of parijoto fruit can increase spermatozoa motility of male Sprague Dawley rats.

Effect of Robusta Coffee Extract on Histopathological in Mice Testes Induced with Monosodium Glutamat

Monosodium glutamate (MSG) is a sodium salt from glutamic acid which is currently very popular to be used as a food flavoring ingredient to stimulate appetite. Exessive consumption of MSG can disrupt the balance of antioxidants and ROS, and cause the negative effects of oxidative stress on testes. Coffee has high chlorogenic acid, function for antioxidants and reduce the negative effect on cell damage in the testes. This study aims to analyze the effect of robusta coffee extract on histological of mice testes induced with MSG. This study was completely randomized design (CRD), using 20 male mice for 5 treatment groups: K+ (MSG 0.12 mg and Vitamin C 6 mg), K- (MSG 0.12 mg and CMC Na 0.1 ml), P1 (MSG 0.12 mg and RCE 0.1 mg), P2 (MSG 0.12 mg and RCE 0.2 mg), and P3 (MSG 0.12 mg and RCE 0.4 mg) orally for 42 days. Histophatological scores were analyzed with Saphiro-Wilk Test and Anova. There is no effect of increasing testicular weight. 0,12 mg MSG can cause a decrease in the number of spermatogonia and spermatocytes. 0,2 mg robusta coffee extract can maintain the number of spermatogonia and spermatocytes.

Toxicological evaluation of a Nigeria-made polyherbal product on selected reproductive functions in adult male Wistar rats

Ruzu bitters black for men (RBBM) is a polyherbal product widely used amongst men in Nigeria to enhance libido, rejuvenate male organs and to manage erectile dysfunctions, prostate anomalies, weak erection, and premature ejaculation. This study was carried out to investigate the toxicological effect from the use of herbal product. Acute toxicity test of RBBM on rats was carried out in two phases; 10 mg/kg, 100 mg/kg and 1000 mg/kg for phase I and 1600mg/kg, 2900 mg/kg and 5000 mg/kg for phase II, were administered respectively. For sub-acute toxicity, two groups of 5 animals each received RBBM (0.87 mg/kg and1.17 mg/kg respectively) and a third group received water orally for 28 days. The study analyzed the median lethal dosage, and sperm morphology, sperm motility, sperm count, sperm viability and histology of the testes as indices for sub-acute toxicity. No death was recorded for the acute and sub-acute studies but there was a moderate physical sign of toxicity. In the sub-acute toxicity study, there was a significant increase (p˂0.05) in testicular weight of Group 1 animals. Also, sperm count, and sperm motility increases significantly (p˂0.05) while there was a decrease in multiple tail sperm across the test groups. RBBM is not toxic to sperm morphology and causes no death at 5000 mg/kg in male albino Wistar rats.

Quercetin ameliorates testosterone secretion disorder by inhibiting endoplasmic reticulum stress through the miR-1306-5p/HSD17B7 axis in diabetic rats

Testicular damage and testosterone secretion disorder are associated with diabetes mellitus. Quercetin, a common flavonoid, has antioxidant, anti-cancer, and blood sugar lowering effects. Therefore, this study aims to investigate the effect of quercetin on the reproductive system of male rats with diabetes in vivo and in vitro and elucidate its mechanism. Streptozotocin (STZ) induction was used to establish a diabetes model in forty male Sprague Dawley (SD) rats, which were subsequently administered with 20 or 50 mg/kg of quercetin. Leydig cells of rat testes were treated by high glucose (HG) followed by 5 or 10 μM quercetin. Two doses of quercetin increased rat body weight and testicular weight, decreased blood glucose,and inhibited oxidative stress. RT-qPCR and Western blotting revealed that quercetin alleviated STZ-induced testicular damage and promoted testosterone synthesis. Both doses of quercetin reduced ROS and MDA levels, and increased SOD level in HG-treated cells. Both, in vivo and in vitro results confirmed that a high dose of quercetin was more effective. MiR-1306-5p was upregulated in testicular tissue of diabetic rats and HG-treated cells. 17β-hydroxysteroid dehydrogenase (HSD17B7) was a target of miR-1306-5p and HSD17B7 was downregulated in STZ-induced rat tissues and HG-treated cells. HSD17B7 overexpression reversed the increase of C/EBP homologous protein (CHOP) and glucose-regulated protein 78 (Grp78) protein levels as well as eIF2α phosphorylation level and promotion of cell apoptosis caused by miR-1306-5p overexpression. Moreover, overexpression of HSD17B7 activated the Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) axis in HG-treated cells. In conclusion, quercetin inhibits ER stress and improves testosterone secretion disorder through the miR-1306-5p/HSD17B7 axis in diabetic rats.

Efek Serbuk Kunyit dan Kurkumin pada Spermatogenesis Mencit (Mus musculus) yang Diberi Minuman Beralkohol

Kunyit mengandung senyawa kurkumin yang dapat digunakan sebagai zat antiinflamasi dan membantu memperbaiki sel-sel yang rusak. Tujuan dari penelitian ini menganalisis pengaruh serbuk kunyit dan kurkumin pada jumlah dan ukuran sel spermatogonium; spermatosit primer; dan spermatosit sekunder; bobot testis serta diameter tubulus seminiferus Mus musculus yang diberi minuman beralkohol. Penelitian ini merupakan penelitian eksperimental dengan Rancangan Acak Lengkap (RAL), menggunakan 12 ekor Mus musculus jantan yang dibagi kedalam 4 kelompok perlakuan dan 3 kali ulangan. R0 merupakan kontrol, R1 kontrol alkohol, R2 pemberian serbuk kunyit sebanyak 0,1 mg/hari, R3 pemberian kurkumin sebanyak 0,01 mg/hari. Perlakuan diberikan selama 30 hari. Data penelitian dianalisis menggunakan analysis of variance (ANOVA) pada taraf kepercayaan 95%. Berdasarkan hasil penelitian yang diperoleh dapat disimpulkan bahwa tidak terdapat perbedaan bermakna (p>0.05) pada jumlah spermatogonium dan ukuran sel (spermatogonium, spermatosit primer, dan spermatosit sekunder), namun terdapat perbedaan bermakna pada (P<0,05) pada bobot testis, diameter tubulus seminiferus dan jumlah sel (spermatosit primer, dan spermatosit sekunder). Turmeric contains curcumin compounds that can be used as anti-inflammatory substances and help repair damaged cells. The purpose of this study was to analyze the effect of turmeric powder and curcumin on the number and size of spermatogonia cells; primary spermatocytes; and secondary spermatocytes; testicular weight and diameter of the seminiferous tubules of Mus musculus given alcoholic beverages. This study is an experimental study with a completely randomized design (CRD), using 12 male Mus musculus which were divided into 4 treatment groups and 3 replications. R0 is control, R1 is alcohol control, R2 is 0.1 mg/day of turmeric powder, R3 is 0.01 mg/day of curcumin. The treatment was given for 30 days. The research data were analyzed using analysis of variance (ANOVA) at the 95% confidence level. Based on the results obtained, it can be concluded that there is no significant difference (p>0.05) in the number of spermatogonia and cell size (spermatogonia, primary spermatocytes, and secondary spermatocytes), but there is a significant difference (P<0.05) in testicular weight, diameter of the seminiferous tubules and the number of cells (primary spermatocytes, and secondary spermatocytes).

Riboflavin Recovery of Spermatogenic Dysfunction via a Dual Inhibition of Oxidative Changes and Regulation of the PINK1-Mediated Pathway in Arsenic-Injured Rat Model

Arsenic trioxide (As2O3) poisoning and associated potential lesions are of a global concern. Inversely, riboflavin (vitamin B2) as a component of flavoproteins could play a vital role in the spermatogenic enzymatic reactions. Thus, this research aimed to explore potential beneficial roles of vitamin B2 during As2O3-injured-toxicity. Rats were randomly allocated into 4 groups (n=8/group) and challenged as follows (for 30 days continuously): Group 1 received normal saline, Group 2 was treated with 3 mg/l As2O3, Group 3 received 40 mg/l vitamin B2, Group 4 received 3 mg/l As2O3 + 40 mg/l vitamin B2. Both As2O3 and vitamin B2 were dissolved in deionized water. Malondialdehyde (MDA), Glutathione Peroxidase (GSH-Px), Superoxide dismutase (SOD), and Catalase (CAT) were assessed for the oxidative profile, while TAS (Total Antioxidative Status) levels were evaluated for the antioxidant system, in both serum and testicular tissue. P<0.05 was considered statistically significant. The results show that As2O3 significantly decreased the body weight, testicular weight and testis volume, semen quality and testicular cell count (p<0.05). Furthermore, MDA content in the testicular tissue of the As2O3 group rats was significantly higher in comparison to the vehicle group (p<0.05). Likewise, TAS and the activities of GSH-Px, CAT and SOD were reduced (p<0.05) when compared to the control. As2O3 induced testicular damage and seminiferous tubular atrophy. Monodansylcadaverine assays mirrored the histopathology observations. Meanwhile, As2O3 upregulated the expression of mitophagy-related genes including PINK1, Parkin, USP8, LC3-I, Fis1 and Mfn2. The p38 gene, responsible to stress stimuli, was also upregulated by As2O3 administration. Meanwhile, exposure to Vitamin B2 led to a significant decrease of the expression levels of mitophagy related genes. Our study revealed that vitamin B2 supplementation protected testicular structures against As2O3-induced injury via a dual inhibition of oxidative changes and a regulation of the PINK1-mediated pathway.

Rubicon prevents autophagic degradation of GATA4 to promote Sertoli cell function

Autophagy degrades unnecessary proteins or damaged organelles to maintain cellular function. Therefore, autophagy has a preventive role against various diseases including hepatic disorders, neurodegenerative diseases, and cancer. Although autophagy in germ cells or Sertoli cells is known to be required for spermatogenesis and male fertility, it remains poorly understood how autophagy participates in spermatogenesis. We found that systemic knockout mice of Rubicon, a negative regulator of autophagy, exhibited a substantial reduction in testicular weight, spermatogenesis, and male fertility, associated with upregulation of autophagy. Rubicon-null mice also had lower levels of mRNAs of Sertoli cell–related genes in testis. Importantly, Rubicon knockout in Sertoli cells, but not in germ cells, caused a defect in spermatogenesis and germline stem cell maintenance in mice, indicating a critical role of Rubicon in Sertoli cells. In mechanistic terms, genetic loss of Rubicon promoted autophagic degradation of GATA4, a transcription factor that is essential for Sertoli cell function. Furthermore, androgen antagonists caused a significant decrease in the levels of Rubicon and GATA4 in testis, accompanied by elevated autophagy. Collectively, we propose that Rubicon promotes Sertoli cell function by preventing autophagic degradation of GATA4, and that this mechanism could be regulated by androgens.