雷公藤多苷致DOR小鼠子宫内膜生长发育及卵巢相关干细胞因子、缝隙连接蛋白表达变化

Changes in endometrial growth and expressions of ovarian related stem cell factors and connexins in DOR mice induced by tripterygium glycosides

目的探讨雷公藤多苷致卵巢储备下降小鼠子宫内膜生长发育及卵巢相关干细胞因子、缝隙连接蛋白表达变化。方法购入小鼠后进行1周适应性喂养,按随机数字表法分为对照组及模型组各10只;对照组予以正常饮食;模型组予以雷公藤制剂灌胃造模,末次给药24小时后,将小鼠进行麻醉取血并处死。计算两组小鼠子宫内膜指数;采用免疫组化法检测小鼠子宫内膜血管内皮生长因子(VEGF)及其血管内皮生长因子受体2(VEGFR2)、卵巢干细胞因子(SCF)、缝隙连接蛋白37(Cx37)的表达;采用双抗体夹心检测法检测小鼠黄体生成素(LH)、促卵泡生成素(FSH)、雌二醇(E2)水平。结果显微镜下观察,模型组卵巢萎缩,各级卵泡数量减少,颗粒细胞排列紊乱,黄体较少,可见病理性纤维化增生及炎性浸润;模型组小鼠子宫指数较对照组降低(t=3.00,P=0.01);模型组小鼠子宫内膜VEGF和VEGFR2的表达水平较对照组均明显降低,差异有统计学意义(t=3.86,P<0.01;t=111.80,P<0.01);模型组小鼠SCF的表达水平较对照组升高,Cx-37表达水平下降,差异有统计学意义(t=14.57,P<0.01;t=27.95,P<0.01);模型组小鼠血液中FSH、LH水平均高于对照组(t=15.40,P<0.01;t=23.90,P<0.01);模型组小鼠血液中E2水平低于对照组,差异有统计学意义(t=12.92,P<0.01)。结论雷公藤多苷可引起小鼠体内Cx-37表达下降,SCF表达上升,导致小鼠卵巢储备功能下降,抑制子宫内膜血管生成。

Objective To explore changes in growth and development of endometrium and expressions of ovarian-related stem cell factor(SCF)and connexin(Cx-37)of mice with decreased ovarian reserve(DOR)caused by Tripterygium wilfordii polyglycosides.Methods 20 female mice bought were fed for one week of adaption and then they were randomly divided into control group and model group in number table method(n=10 in each group).The mice in the control group were given normal diet,and those in the model group were given Tripterygium wilfordii preparation by gavage.24 hours later after the last administration,the mice were anesthetized and killed.The uterine index was caculated and expressions of VEGF,VEGFR2,SCF and Cx-37 in endometrium of the mice were detected by immunohistochemistry method.The serum levels of LH,FSH and E2 of the mice were detected by double antibody sandwich assay method.Results Under the microscope,atrophied ovaries,reduced number of follicles at all levels,disordered arrangement of granulosa cells,less number of corpus luteum,and pathological fibrosis proliferation and inflammatory infiltration of the mice in the model group could be seen.The uterine index of the mice in the model group was lower than that in the control group(t=3.00,P=0.01).The expression levels of VEGF and VEGFR2 in endometrium of the mice in the model group were significantly lower than those in the control group,there were significant differences between the two groups(t=3.86 and 111.80 respectively,both P<0.01).The expression level of SCF in the model group was higher than that in the control group,while the expression level of Cx-37 was lower(t=14.57 and 27.95 respectively,both P<0.01).The serum levels of FSH and LH of the mice in the model group were higher than those in the control group,there were significant differences between the two groups(t=15.40 and 23.90 respectively,both P<0.01).The serum level of E2 of the mice in the model group was lower than that in the control group and there was a significant difference between the two groups(t=12.92,P<0.01).Conclusions Tripterygium wilfordii polyglycosides can reduce expression of Cx-37 and increase expression of SCF in mice,resulting in decrease of ovarian reserve function and inhibition of endometrial angiogenesis.