Blepharis persica increases testosterone biosynthesis by modulating StAR and 3β-HSD expression in rat testicular tissues

Objective: To evaluate the effect of methanolic extract and ethyl acetate fraction of methanol extract prepared from the seeds of Blepharis (B.) persica on testosterone biosynthesis and also to elucidate the underlying mechanism. Methods: Forty-eight male Wistar rats were divided into eight groups (n=6 per group). Group I received 0.3% w/w gum acacia suspension p.o. and served as the normal control group. Group II was administered testosterone propionate in arachis oil i.m. as the positive control group. Group III to V received B. persica methanolic extract p.o. at doses of 50, 100 and 200 mg/kg body weight. Group VI to VIII received B. persica ethyl acetate fraction p.o. at doses of 50, 100 and 200 mg/kg body weight. The testis was used for biochemical estimation and histological studies. The effects of methanolic extract and ethyl acetate fraction of B. persica on testicular testosterone, mRNA expression corresponding to steroidogenic acute regulatory protein (StAR) and 3β-hydroxysteroid dehydrogenase (3β-HSD) along with 3β-HSD enzyme assay were evaluated in testicular tissues and sperm concentration. Ethyl acetate fraction of B. persica was subjected to column chromatography. In-vitro studies were performed using TM3 cell line at three dose levels (50, 100, 200 μg/mL), each for methanolic extract, ethyl acetate fraction and 2-benzoxazolinone for evaluation of their comparative effect on testosterone production. Results: Ethyl acetate fraction and methanolic extract of B. persica could elevate the testicular testosterone content compared to the normal control group. The treatment with methanolic extract and ethyl acetate fraction of B. persica increased the expression of mRNA corresponding to StAR by 6.7 fold and 10.6 fold, respectively, whereas the mRNA expression of 3β-HSD increased by 5.7 fold and 7.3 fold, respectively. Moreover, fraction and extract treatment exhibited increased 3β-HSD activity in the testicular tissues and were found to elevate sperm concentration in seminal fluid. The spermatogenic potential was further ensured by histological observations. 2-benzoxazolinone was isolated from ethyl acetate fraction and identified using spectral studies. It showed the ability to increase the testosterone content in the TM3 Leydig cells. Conclusions: Methanolic extract and ethyl acetate fraction of B. persica are able to increase the testicular testosterone in rats by elevating mRNA expression of StAR and 3β-HSD in testicular tissues, leading to increase the sperm concentration.