目的:研究原癌基因c-src在大鼠卵巢的表达,及其在原始卵泡启动过程中的作用。方法:取2日龄SD雌性大鼠卵巢,在Waymouth培养体系中培养0.4、8d后,首先采用RT-PCR方法证实大鼠卵巢中有c-src的表达,再体外合成其RNA小干扰片段(small interference RNA,siRNA)转染培养中的卵巢组织进行RNA干扰,用HE染色及RT-PCR筛选最佳干扰片断并用慢病毒包装后检测干扰效果。结果:随着培养天数的增加,原始卵泡在卵泡总数中所占比例逐渐减少;c—src mRNA在原始卵泡中有表达,经筛选用最佳干扰片断siRNA1慢病毒包装进行RNA干扰,发现干扰后,与空白组、空白载体组相比,最佳干扰组c—src mRNA含量明显下降,原始卵泡在卵泡总数中所占比例相对更多,原始卵泡发育受到抑制。结论:c-src在原始卵泡中有表达,并在一定程度上促进了原始卵泡的发育。
Objective: To explore the role of c-src on the initiation of primordial follicles. Methods: 2-days-old female SD rats' ovaries were cultured in Waymouth culture system and were used HE staining and immunohistochemy to observe the number of follicles after 0,4,8 days cultured. Use chemically synthesized small interference RNA(siRNA)transfected into ovarian tissue in cultured for RNA interference, and use HE staining and RT-PCR to detect the best siRNA and packaging it by lentiviruses to test the interference effect. Results: With the increase of culturing days, the nummber of the primordial follicles in ovarian gradually reduced. We packed the best siRNA by lentiviruses to doing RNA interference and found comparing with the blank control group and blank vector group, c-src mRNA of the best interference group were significantly decreased, the total number of primordial follicles was relatively greater and the development of primordial folliculars was inhibited. Conclusion: c-sre plays an important role in primordial follicle development and folliculogenesis initiation.