The present study aims to investigate the roles of steroidogenic acute regulatory protein (StAR) in Yangjing Capsule (YC) induced anti-apoptotic effects on Leydig cells and the related mechanism. Leydig tumor cells (MLTC-1) were cultured and treated with YC, and immunofluorescence assay was performed to examine the expression of StAR; furthermore, luciferase reporter assay was conducted to evaluate the impact of YC on StAR promoter; next, MLTC-1 cells were treated with StAR small interfering RNA (siRNA), and flow cytometry was carried out to examine the effect of StAR siRNA on the apoptosis of the cells; furthermore, quantitative (q)RT-PCR and Western blot methods was used to determine the expression of StAR and apoptosis related molecules Bcl-2, Bax and Caspase-3 on both mRNA and protein levels in different groups; finally the secretion of testosterone in different groups was examined by radioimmunoassay. We observed that the YC can increase the expression of StAR in a dose-dependent manner, and YC can activate the promoter of StAR; moreover, transfection of StAR siRNA can block YC induced anti-apoptotic effects and increased production of testosterone. In conclusion, our results suggested that YC might suppress the apoptosis of MLTC-1 cells and enhance the production of testosterone through regulating the expression of StAR.

Purpose: To explore the protective effects of Yangjing capsule (YC) on testicular microcirculation in a mouse model of deficiency of testicular microcirculation. Methods: Immunohistochemistry was applied to determine the effects of YC on microvascular density of mice. The protein level of CD34 and vascular endothelial growth factor A (VEGF A) was measured by western blot. The viability of Testicular cell line (TM4 cells) was examined by CCK-8 assay. Results: Histopathological changes demonstrated that CP-induced decrease of microvascular density of the mice was rescued by YC dose-dependently (p < 0.5). Western blot data showed that the protein levels of CD34 and VEGF A in CP group were significantly decreased, but dose-dependently increased by YC, respectively, following co-administration of CP + YC, compared with those in CP group (p < 0.5). The results from CCK-8 assay showed that the cell viability of TM4 cells increased with the amount of YC administered, and that high concentrations of YC (0.1 and 1 mg/mL) showed significant effects (p < 0.5). Moreover, YC showed little effect on VEGF A mRNA and protein expression in TM4 cells. Conclusion: YC may be considered an alternative therapeutic agent for the management of testicular microcirculation disease. However, further studies are required to ascertain this. Keywords: Yangjing Capsule, Testicular microcirculation, Cyclophosphamide, Vascular endothelial growth factor A

Background. The testicular microcirculation was an important aspect of testicular physiology and it offered a stable environment for the transport of nutrients and secretary products in the testis. Yangjing capsule (YC), a traditional Chinese compound herbal prescription, has been proved as an effective drug to ameliorate spermatogenesis, promote testosterone synthesis in vivo, and cure spermatogenesis in clinical practice. Objective. This study was aimed at understanding the potential mechanisms of YC exerting angiogenic effects in the mouse spermatogenesis dysfunction model induced by cyclophosphamide (CP) and MLTC-1 cells. Materials and Methods. Balb/c mice were randomly divided into five groups: control, CP, CP plus YC (630 mg/kg), CP plus YC (1260 mg/kg), and CP plus YC (2520 mg/kg). After 30 days, mice were sacrificed and the expressions of endothelial marker CD34+, angiogenic marker VEGFA, VEGFR1, VEGFR2, and eNOS in the testes of the mice were examined; moreover, Leydig cell line MLTC-1 cells were cultured and treated with different concentrations of YC extracts (YCE), and the expressions of VEGFA, VEGFR1, VEGFR2, and eNOS, as well as the secretion of NO, were evaluated. Results. We observed that YC significantly increased the expressions of VEGFA, VEGFR1, VEGFR2, and eNOS in testes of CP-treated mice; moreover, YCE has led to increased expressions of VEGFA, VEGFR1, VEGFR2, and eNOS and secretion of NO in MLTC-1 in vitro. These data suggested that the YC might be an alternative treatment for the dysfunction of testicular microcirculation by promoting the angiogenesis in the testis.