补肾健脾方对自然衰老小鼠卵巢端粒的调控作用

Effects of Bushen Jianpi Formula on ovary telomeres in natural aging mice

目的:观察自然衰老小鼠模型和端粒酶的相关性以及补肾健脾方对自然衰老小鼠模型端粒调控作用的影响。方法:BALB/c小鼠按月龄分为正常组、模型组、补肾健脾方组,分别给予药物或蒸馏水,干预4周后,取小鼠血清,卵巢。放射免疫法检测小鼠血清雌二醇(E2)的水平,HE染色观察卵巢组织病理形态学改变,TUNEL法检测组织细胞凋亡,Real Time PCR方法检测卵巢端粒酶活性以及端粒的相对长度。结果:与正常组比较,模型组小鼠E_(2)水平、端粒酶活性及端粒相对长度显著降低(P<0.05,P<0.01),卵巢颗粒细胞凋亡率显著升高(P<0.05);与模型组比较,补肾健脾方组E_(2)水平、端粒酶活性及端粒相对长度显著升高(P<0.05,P<0.01),卵巢颗粒细胞凋亡率显著降低(P<0.05)。模型组卵巢闭锁卵泡数量多于正常组,补肾健脾方组卵巢中的生长卵泡和成熟卵泡数量多于模型组。结论:补肾健脾方通过提高雌激素水平,促进卵泡的发育,抑制卵巢颗粒细胞的凋亡,而且可以增加卵巢端粒酶的活性和端粒长度,进而抑制小鼠卵巢衰老。

Objective: To investigate the correlation between telomerase and natural aging mouse model and the effects of Bushen Jianpi Formula on telomere regulation in natural aging mouse model. Methods: BALB/c mice were divided into normal group, model group and Bushen Jianpi Formula group. Mice were administrated drugs or normal saline by gastric gavage, and serum and ovaries were collected after intervention for four weeks. The serum E2 levels were detected by radioimmunoassay,ovarian histopathological changes were observed by HE staining, tissue apoptosis was detected by TUNEL assay, and the changes in telomerase activity and telomere relative length in ovarian tissue samples were detected by Real-Time PCR. Results:Comparison of the normal group, the estradiol level, the telomerase activity and telomere relative length in the natural aging model group were significantly decreased(P<0.05, P<0.01), and the apoptosis rate of ovarian granulosa cells was significantly increased(P<0.05). Compared with model group, the estradiol level, the telomerase activity and telomere relative length in the Bushen Jianpi Formula group were significantly decreased(P<0.05, P<0.01), and the apoptosis rate of ovarian granulosa cells was significantly decreased(P<0.05). The model group had more atretic follicles than the normal group. There were more growing follicles and mature follicles in the ovaries of the treated group than that in the model group. Conclusion: Bushen Jianpi Formula had estrogenlike effects to inhibit the effect of ovarian aging in mice by promotion of the development of follicles, inhibition of the apoptosis of ovarian granulosa cells, and increasement of the activity of ovarian telomerase and telomere length.