不同浓度补肾健脾方对卵巢颗粒细胞增殖作用的影响

The Influence of Kidney-Supplementing and Spleen-Activating Decoction with Different Concentrations on Ovarian Granulosa Cell proliferation

目的:观察不同浓度补肾健脾方含药血清对原代培养的卵巢颗粒细胞增殖作用的影响以及不同浓度补肾健脾方对小鼠卵巢组织卵泡颗粒细胞层增殖的影响。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染色法进行细胞形态鉴定。采用MTT法检测,分别观察含质量分数5%、10%、20%补肾健脾方含药血清对卵巢颗粒细胞增殖作用的影响。体内实验通过给雌性小鼠灌胃不同浓度的补肾健脾方,连续灌胃4周,取卵巢组织做病理形态学观察。结果:质量分数20%、10%、5%的补肾健脾方含药血清对大鼠卵巢颗粒细胞增殖作用各不相同,质量分数20%、10%的含药血清、20%的空白血清对卵巢颗粒细胞的增殖作用最明显,与正常组比较,差异具有统计学意义(P<0.05)。补肾健脾方大剂量、中剂量、小剂量对卵巢卵泡颗粒细胞层的厚度呈现一定的增高趋势。结论:补肾健脾方对小鼠的卵巢颗粒细胞具有一定的增殖作用。

Objective： To observe the influence of serum in Kidney-Supplementing and Spleen-Activating Decoction with different concentrations on the proliferation of primary cultured ovarian granulnsa cells, and the influence of Kidney-Supplementing and Spleen-Activating Decoction with different concentrations on the proliferation of grannlosa cells in the ovary of rats. Methods ： The primary cultured rat granulosa cells were cultured in vitro by mechanical separation and trypsin digestion methods. The cell mor- phology was identified by HE staining. MTT method was used to observe the influence of serum in Kidney-Supplementing and Spleen-Activating Decoction with 5% , 10% ,or 20% concentrations on the proliferation of ovarian granulosa ceils. In vivo experi- ment,the mice were fed with different concentrations of Bnshen Jianpi Decoction for 4 weeks. Ovarian tissue was observed by pathological morphoiogy. Results. The serum in Kidney-Supplementing and Spleen-Activating Decoction with 20%, 10%, or 5% concentration had different influence on rat granulosa cell proliferation. Medicated serum with 20%, or 10% concentration, the blank serum with 20% concentration had the most obvious effect on the proliferation of ovarian granulosa cells. Compared with the normal group, the differences were statistically significant （ P 〈 0.05 ）. The high dose, medium dose and small dose of Kidney-Supplementing and Spleen-Activating Decoction had certain tendency of increasing the thickness of granulosa cell layer. Conclusion ： Kidney-Supple menting and Spleen-Activating Decoction has a certain effect on proliferation of mice＇s ovarian granulosa cells.