补肾健脾方对卵巢颗粒细胞激素水平和细胞周期的影响

Effect of Bushen Jianpi on Ovarian Granulosa Cell Hormone Levels and Cell Cycle

目的通过卵巢颗粒细胞原代培养,观察补肾健脾方含药血清对原代培养的卵巢颗粒细胞激素水平和细胞周期的影响。方法采用机械分离法结合胰蛋白酶消化法体外原代培养大鼠卵巢颗粒细胞,通过HE染色进行细胞鉴定。采用放免法检测E_2的水平,并用流式细胞仪检测细胞周期的变化。结果补肾健脾方含药血清对卵巢颗粒细胞分泌物中的E_2水平明显升高,补肾健脾方含药血清大剂量组、中剂量组、小剂量组卵巢颗粒细胞分泌物中的E_2水平与空白血清组比较具有显著性差异(P<0.01,P<0.05)。补肾健脾方干预后卵巢颗粒细胞的G_2、S期所含的细胞比例有明显的影响,G_2期所含的细胞比例明显降低,与正常细胞比较具有显著性差异(P<0.05),S期所含的细胞比例明显升高,与正常细胞比较具有显著性差异(P<0.05)。结论补肾健脾方含药血清可促进卵巢颗粒细胞由G0期向S期转化,从而促进细胞的增殖。并且具有明显的升高原代培养的卵巢颗粒细胞的E_2水平的作用。

Objective Observe the effects of kidney and spleen contained serum on primary cultured ovarian granulosa cells hormone levels and cell cycle. Methods Using mechanical separation method combined with trypsin digestion method in vitro primary culture of rat ovarian granulosa cells, cell identification by HE staining. Put free method is used to measure the levels of E2, and the change of flow cytometry instrument to detect the cell cycle. Results Bushen Jianpi drug-containing serum E2 level in ovarian granular cell secretion increased significantly. Bushen Jianpi drug-containing serum large dose group, middle dose group and small dose group of ovarian granular cell secretion E2 level compared with the blank serum group with significant difference （P 〈 0.01, P 〈 0.05）.Prognosis ofBushen Jianpi drug-containing serum, ovarian granulosa ceils ofG2, S phase cell proportion have the obvious effect, significantly lower proportion of cells in G2 phase that compared with normal cells with significant difference （P 〈 0.05）. S period, the proportion of cells with increased significantly, compared with normal cells with significant difference （P 〈 0.05）. Conclusion Bushen Jianpi drug-containing serum can promote ovarian granulosa cells from Go to S phase transformation, thus promotes cell proliferation.And it has the obvious rise plateau generation of cultured ovarian granulosa cells E2 levels.