科研之友
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摘要
Follistatin (FST), a local regulator of gonadal functions is a powerful inhibitor of follicle stimulating hormone (FSH) secretion. In the present study, the expression of FST was partially silenced at both transcriptional and translational levels by RNAi-Ready pS1REN-RetroQ-ZsGreen Vector mediated recombinant pshRNA vectors in bovine granulosa cells (bGCs). The results showed that transfection with FST-1 and FST-2 vectors significantly down-regulated mRNA and protein expressions of follistatin by 51% (P=0.0093) and 72% (P=0.0078) respectively. After down-regulation of FST in bGCs, cell cycle was arrested at S-phase (9.2 +/- 0.6 vs 12.5 +/- 0.2, P=0.0055), and apoptosis was significantly (21.3 +/- 2.7 vs 13.9 +/- 2.5, P=0.0051) increased. These findings were further verified by down-regulation of protein level of B-cell leukemia/lymphoma 2 (Bcl2, P=0.0423), and up-regulation of caspase-3 (P=0.0362), p21 (P=0.0067) and mRNA levels of Bcl2-associated X protein (Bax, P=0.041). Knockdown of FST in bGCs significantly increased activin A concentration in culture medium, while level of estradiol (E2) was suppressed without affecting progesterone production. In addition, mRNA levels of all activin receptor subtypes [activin receptor types I (ACRI) and II (ACRIIA and ACRIIB)] and inhibin alpha-subunit were augmented (P<0.05) without altering both inhibin beta-subunits. These findings suggest that follistatin may participate in caspase3-dependent apoptosis through Bcl2/Bax gene family in bovine GCs, whereas, activin and its receptors are associated with its regulation. Activin-induced up-regulation of inhibin-alpha subunit in bGCs seems to be involved in the regulation of steroidogenesis.
- 出版日期2015-4
- 单位华中农业大学
