二仙汤对顺铂所致大鼠卵巢早衰模型中卵巢颗粒细胞增殖及周期的影响

Influences of Erxian decoction on proliferation and cell cycle of ovarian granulosa cells in premature ovarian failure model caused by CDDP

目的观察二仙汤对顺铂损伤大鼠卵巢颗粒细胞增殖及细胞周期的影响。方法以不同剂量的二仙汤药理血清作用于体外培养的顺铂损伤卵巢颗粒细胞,并以正常药理血清为空白对照,戊酸雌二醇药理血清为阳性对照,同时对比加入PI3K/AKT通路的抑制剂LY294002后各组的变化。MTT法检测各组药理血清作用于卵巢颗粒细胞24小时、48小时和72小时的增殖情况。流式细胞术检测各组中细胞周期各时相的变化。结果 (1)二仙汤对顺铂损伤后的卵巢颗粒细胞具有增殖促进作用(P<0.05);且对于顺铂损伤后并给予LY294002抑制剂的颗粒细胞也具有增殖改善作用(P<0.05)。(2)二仙汤各剂量组及戊酸雌二醇组均能使处于S期的顺铂损伤颗粒细胞比例显著增多,G_0/G_1期细胞相对比例减少(P<0.01),且增殖指数升高(P<0.01)。结论二仙汤对卵巢颗粒细胞增殖有显著促进作用,其作用机制可能是激活或增强AKT信号通路,抑制细胞凋亡,从而促进细胞增殖;另一方面,也可能是通过促进颗粒细胞从G_0期向S期转化,增加S期细胞比率来实现的。

Objective To observe the influences of Erxian decoction on the proliferation and cell cycle ovarian granulosa cells caused by CDDP. Methods Different doses of Erxian decoction pharmacological serum were injected in ovarian granulosa cells injured by CDDP,normal pharmacological serum was used as blank control,progynova pharmacological serum was used as positive control,and focus on the comparison of changes after joined LY294002 which was PI3 K / AKT pathway inhibitor in each group at the same time. Each group of ovarian granulosa cells proliferation was detected by MTT method when pharmacological serum dosing for 24 h,48h and 72 h. Cell cycle and phase changes in each group were determined by flow cytometry. Results（ 1） Erxian decoction has proliferation promoting effect on CDDP injury ovarian granulosa cells,and has proliferation effect of ovarian granulosa cells after injured by CDDP and added LY294002 inhibitors.（ 2） Erxian decoction each dose group and progynova group can rise the proliferation index by enhancing the proportion of S phase granulosa cells and decreasing G0/ G1 phase proportion. Conclusions Erxian decoction has significant proliferation promoting effect of CDDP injuryovarian granulosa cells. The possible mechanism on one hand is that it activates or enhances AKT signaling pathways and inhibits cell apoptosis. On the other hand,it perhaps promotes S phase proportion transformation from G0 phase therefore increases the ratio of S phase cells.