大鼠子宫内膜慢性缺血引起子宫内膜端粒酶和c-kit表达减少

Decrease of endometrial thickness and gland number and decreased expression of telomerase and c-kit in rats with chronic endometrial ischemia

目的观察大鼠子宫内膜慢性缺血后子宫内膜厚度、子宫内膜腺体数目的变化、同时检测端粒酶(TERT)、POU5f1、c-kit和caspase3的表达。方法实验组通过结扎8周龄SD大鼠双侧子宫动脉构建慢性子宫内膜缺血性损伤模型(实验组),对照组为假手术组。通过阴道细胞涂片判定性周期。3个月过后,于发情期处死SD大鼠,取子宫内膜组织HE染色比较实验组和对照组子宫内膜厚度、腺体数量;免疫组织化学技术检测端粒酶和caspase3的表达;荧光定量PCR检测c-kit、POU5f1和caspase3的表达;Western blot检测端粒酶、c-kit和caspase3的表达情况。结果和对照组比较,模型组内膜厚度(502±148)、腔上皮厚度(17.20±4.98)、腺上皮厚度(9.17±1.96)、内膜腺体数目(18.9±4.8)以及内膜腺体体积分数(0.15±0.02)和对照组内膜厚度(800±130)、腔上皮厚度(29.46±9.27)、腺上皮厚度(15.68±2.50)、内膜腺体数目(30±5.5)以及内膜腺体体积分数(0.40±0.16)比较均显著降低(P<0.05);模型组c-kit mRNA和蛋白表达水平[(0.23±0.01)、(0.31±0.13)]较对照组[(1.00±0.06)、(0.75±0.09)]显著降低(P<0.05);模型组caspase3 mRNA和蛋白表达水平[(1.43±0.22)、(0.67±0.23)]明显高于对照组[(1.00±0.16)、(0.40±0.08),P<0.05],端粒酶蛋白表达水平(0.64±0.19)明显低于对照组[(0.87±0.12),P<0.05]。结论子宫内膜慢性缺血以及子宫内膜慢性缺血引起的端粒酶和c-kit表达下调,caspase3表达增加可能是薄型子宫内膜发病的一个因素。

Objective To investigate the decrease of endometrial thickness and gland number and the decreased expressions of telomerase,POU5f1,c-kit,and caspase3 in rats with chronic endometrial ischemia.Methods The experimental group had 30 8-week-old SD rats with the uterine arteries on both sides ligated to build the animal models of chronic endometrial ischemia,as contrasted with a control group（Sham group）.The sexual cycles of the rats were determined by vaginal smear method.The SD rats were killed when entering estrus after 3 months.The endometrial tissue of the rats was collected and treated by HE staining to compare the experimental group and control group in terms of endometrial thickness and gland number.The expressions of telomerase and caspase3 were investigated by immunohistochemistry,the expressions of c-kit,POU5f1,and caspase3 by fluorescence quantitative PCR,and the expressions of telomerase,C-kit,and caspase3 by Western blotting.Results The endometrial thickness（502±148）,luminal epithelium thickness（17.20±4.98）,gland epithelium thickness（9.17±1.96）,and gland number（18.9±4.8）in the experimental group were significantly lower than those in the control group（800±130,29.46±9.27,15.68±2.50,and 30±5.5,respectively）（P0.05）.The expressions of c-kit mRNA and protein（0.23±0.01 and 0.31±0.13） in the experimental group were significantly lower than those in the experimental group（1.00±0.06 and 0.75±0.09）（P0.05）.The expressions of caspase3 mRNA and protein（1.43±0.22 and 0.67±0.23） in the experimental group were significantly higher than those in the control group（1.00±0.16 and 0.40±0.08）（P0.05）.The expression of telomerase protein（0.64±0.19） in the experimental group was significantly lower than that in the control group（0.87±0.12）（P0.05）.Conclusion Chronic endometrial ischemia and its associated expression decrease of c-kit and telomerase and expression increase of caspase3 may be a factor in the pathogenesis of thin endometrium.