党参炔苷对雌性大鼠卵巢颗粒细胞增殖分化的影响及其作用机制

Effects of lobetyolin on ovarian granulose cells' proliferation,differentiation and the mechanism

目的:探讨党参炔苷对雌性大鼠卵巢颗粒细胞(GCs)增殖分化的影响及其作用机制。方法:观察大鼠卵巢GCs的生长状况,通过CCK-8实验和细胞雌二醇(E2)分泌量测定确定细胞体外培养时间以及最佳给药浓度,进一步观察党参炔苷对体外培养的卵巢GCs增殖分化及分泌功能的影响,并通过阻断GCs中的c AMP信号通路、ERK 1/2信号通路、p38MAPK信号通路和钙离子(Ca2+)通道来探究其对卵巢GCs功能影响的作用机制。结果:确定24 h为GCs最佳共培养时间;党参炔苷浓度为3.125 mg/L时,能显著增加E2的分泌量(P<0.01),对细胞3β-羟基固醇脱氢酶(3β-HSD)含量和CCK-8实验吸光度(D)值均无影响(P>0.05)。c AMP通路抑制剂H89和p38MAPK通路抑制剂SB203580能够抑制党参炔苷对卵巢GCs分泌E2的促进作用(P<0.01)。结论:党参炔苷可有效促进卵巢GCs分泌E2,且不影响细胞的增殖和正常分化;党参炔苷可能通过c AMP和p38MAPK信号通路来促进GCs对E2的合成及分泌。

Objective： To investigate the effects of lobetyolin on proliferation, differentiation of ovarian granulosa ceils （GCs） and its mechanism. Methods： Ovarian GCs＇ growth, tested by cell counting experiment （CCK-8） and the estradiol （E2）＇s secretion was deserved to determine the time of rats＇ ovarian GCs cultured in vitro and the optimal dosing concentration so that to research the effect of lobetyolin on ovarian GCs＇ proliferation, differentiation and secretion function. At the same time, by blocking cAMP signal pathways, ERK1/2 signal pathways, p38MAPK signal pathways and calcium ion channels, the mechanism of the influence on ovarian GCs＇ secretion function was explored. Results： Twenty-four hours was the optimal culture time for cells. When the concentra- tion of lobetyolin was 3.125 mg/L, it significantly increased the secretion of E2 （P〈0.01）. Compared with the control, the absorbance values of CCK-8 experiments and the content of 3β-HSD were not significantly different （P〉0.05）. H89 and SB203580 could resist the effect of lobetyolin on GCs （P〈0.01）. Conclusion： Lobetyolin could promote ovarian GCs to secrete E2, possibly through the cAMP and p38MAPK signal pathways. At the same time, lobetyolin didn＇t affect the GCs＇ differentiation and proliferation.