补益阳明津气延缓初老雌性大鼠卵巢机能衰老的机理研究

【作者】 李燕；

【导师】 谭万信；

【作者基本信息】 成都中医药大学 ， 中医妇科学， 2004， 博士

【摘要】 研究目的：探讨“补益阳明津气”延缓初老雌性大鼠卵巢机能衰老的机理，并以此佐证中医传统经典理论“五七，阳明脉衰”在绝经前期生殖轴机能衰老中的重要影响。为促进绝经前期妇女健康，延缓绝经前期卵巢机能衰老，防治相关病症，提供一种新的研究与论治思路，并为研制相关方药提供药效学依据。 研究方法：4～6月龄雌性大鼠为正常对照组；10～12月龄，阴道细胞学表现动情期延长的雌性大鼠作为初老大鼠模型。模型动物随机分为：(1)益胃汤高剂量组(2)益胃汤中剂量组(3)益胃汤低剂量组(4)己烯雌酚组(5)模型对照组。灌药4周后，股动脉取血测E2、P、T、FSH、LH；断头处死，取左侧卵巢，HE染色和免疫组化法检测生长卵泡、黄体和卵巢血管数，血管形态以及卵巢ER、FSHR表达，细胞凋亡指标Bcl-2、Bax、Fas、FasL、Caspase3、细胞色素C。部分卵巢作单细胞悬液用于流式细胞凋亡检测；右侧卵巢用于RT-PCR测卵巢抑制素。亚基mRNA的表达。 研究结果：与模型对照组比较，生殖轴内分泌调控方面：补益阳明津气中药复方益胃汤能增加子宫指数、卵巢指数，调整血中性激素水平，改变卵泡生长的激素内环境，促进卵泡生长，使卵巢微血管数目增加，血管壁变薄，管腔变大，使卵巢抑制素α亚基mRNA表达增加，正向调节卵巢的ER和FSHR；卵巢细胞凋亡的研究方面：益胃汤可使卵巢Bcl-2表达增强，Bax表达减弱，Bcl-2／Bax比例增加，Caspase表达减少，细胞色素C减少，流式细胞双标百分比降低，从线粒体途径抑制卵巢细胞凋亡。但未能恢复至正常对照组水平。成都中医药大学2001级博士学位论文 结论:实验研究提示益胃汤具有改善初老雌性大鼠卵泡生长的激素内环境，促进卵泡生长，增加卵巢抑制素的分泌，抑制FSH，改善卵巢血供;提高激素受体表达及抑制卵巢细胞凋亡等作用。这些作用可能是其延缓卵巢机能衰老的机理。更多还原

【Abstract】 Objects: To explore the effect and mechanism of "tonifying Yang Ming Fluid and Qi" to delay the aging of the primary senile rat ovary and so that to testify the importance of TCM traditional classic theory "Yang Ming Meridian Declines during Wu Qi period" in premenopause. The study may offer a new study and differentiational thought to improve premenopausal women health, delay the aging of ovary founction, prevent and treat relative diseases. It also can offer pharmacodynamics evidence for manufacturing relative effective formulas.Methods: 4~6 months old SD female rats were set to be normal control group and 10~12 months old SD female rats whose oestrus elongation were set as primary senile female rat model. Primary senile female rats were devided into 5 groups randomly as following: I. High dose Yiweitang group; 2. Middle dose Yiweitang group; 3. Low dose Yiweitang group; 4. Diethylstilbestrol group; 5. Model control group. The intervention period was 4 weeks. The blood samples were tested by radio-immunity method to detect serum E2, P, T, FSH and LH. The ovaries were treated by HE and Immune histochemistry method to observe growing follicles, corpus luteum, vascular amount, structure of vascular across section, expression of Bcl-2, Bax, Fas, FasL, Caspase3, Cytochrome C. Ovary cells suspension was tested by Flow Cytometry (FCM) to detect apoptosis and the expression of right ovary Inhibin amRNA was measured by RT-PCR.Results: In modulating H-P-O-Ar aspect, compound Yiweitang which could tonify YangMing Fluid and Qi could increase uterus index, ovary index, adjust sex hormone level in blood, change the growth internal environment of follicle, promote the growth of follicle, vascular production of ovary, up regulate the expression of ovary Inhibin amRNA, ER and FSHR. In modulating ovary apoptosis aspect, Yiweitang could improve the expression of Bcl-2 in ovary and weaken the expression of Bax, which could increase the ratio of Bcl-2/Bax. It could also decrease the expression of Caspase3, Cytochrome C and reduce the percentage of apoptosis. Yiweitang could inhibit apoptosis from mitochondrial pathway, while couldn’t reach the normal group level. Conclusions: The results showed that Yiweitang, which could tonify YangMing Fluid and Qi, could change the growth internal environment of follicle, promote the growth of follicle, increase the excretion of ovary inhibin and decrease FSH, improve the blood supply of ovary, increase the expression of hormone receptor and restrain ovary apoptosis. These effects may be the mechanism of Yiweitang delay the aging of primary senile rat ovary.更多还原