目的:研究Sonic Hedgehog(Hh)信号通路关键蛋白Shh、Ptch1、Smo、Gli1在机械损伤后大鼠子宫内膜的表达及意义.方法:将24只8周龄wistar大鼠随机分为正常组(6只)及实验组(18只).实验组大鼠采用机械刮宫法建立大鼠子宫内膜损伤模型,分别于子宫机械损伤后24h、72h和7d各取6只大鼠子宫行苏木精-伊红染色、Masson染色、免疫组织化学染色和实时定量多聚核苷酸链式反应(PCR)检测.结果:子宫内膜机械损伤后24h时大鼠子宫内膜结构破坏,子宫内膜上皮缺失,腺体减少;损伤后72h时大鼠子宫腔被薄层内膜上皮覆盖;损伤后7d子宫内膜较正常内膜薄,腺体明显减少,子宫腔形态差.Masson染色结果显示大鼠子宫内膜机械损伤后7d胶原纤维较正常大鼠子宫内膜明显增加.免疫组化结果显示Shh、Smo、Ptch1及Gli1染色主要位于子宫腺体及内膜周围.Shh在各组大鼠均无表达.Smo在刮宫后24h表达很弱,72h后表达逐渐增强.Ptch1在刮宫后24h为弱表达,72h后表达量明显上升.Gli1在刮宫后72h及7d表达量明显增加.实时定量PCR结果显示Shh mRNA在各组各时间点大鼠子宫均无表达.与正常组子宫内膜相比刮宫后24h Smo mRNA表达量为,0.7倍,Ptch1 mRNA的表达量为1.2倍,Gli1 mRNA表达量为1.6倍(P＜0.05).刮宫后72h,Smo mRNA表达量为1.4倍(P＜0.05),Ptch1 mRNA的表达量为3.3倍(P＜0.01),Gli1 mRNA表达量为9.3倍(P＜0.01).刮宫后7d,Smo mRNA表达量为1.5倍(P＜0.05),Ptch1 mRNA的表达量为2.5倍(P＜0.05),Gli1 mRNA表达量为5.7倍(P＜0.01).大鼠子宫机械损伤后72h Ptch1和Gli1的mR-NA的表达量均高于24h的表达量,且72h Gli1的mRNA表达量显著高于7d的表达量(P＜0.01).结论:Ptch1、Smo、Gli1在机械损伤后大鼠子宫内膜均有表达,Ptch1 、Smo、Gli1的mRNA表达量的变化与大鼠子宫内膜损伤后纤维化修复相关.Hh信号通路可能参与子宫内膜损伤后纤维化修复.%Objective:To investigate the expression and significance of Shh,Ptch1,Smo and Gli1 of Hedgehog signaling pathway in rat endometrium after mechanical injury.Methods:24 eight weeks old wistar rats were randomly divided into normal group (6 rats) and experimental group (18 rats).The rat endometrium injury models in experimental group were established by mechanical curettage.24 hours,72 hours and 7 days after curettage respectively,the uterine specimens of six rats were taken from experimental group and were examined by hematoxylin eosin staining,Masson staining,immunohistochemistry and real time quantitative polymerase chain reaction at different time points.Results:24 hours after endometrial mechanical injury,the endometrial structure of rats was destroyed,the endometrial epithelium was missing and the glands were reduced.72 hours after the mechanical damage of rat uterus,the uterus cavity was covered by thin layer epithelium.7 days after the endometrial mechanical injury,the rat endometrium was slightly thic ker than that at 72 hours,glands were decreased significantly,uterine cavity form was poor,endometrium was still thinner than the normal endometrium.The results of Masson staining showed that 7 days after the mechanical injury of the endometrium,the collagen fibers was significantly increased in the endometrium of rats than that of the normal rats.The results of Masson staining showed that collagen fibers were significantly increased in the endometrium,which were more wounded than those of the normal rats' endometrium at7 days after the mechanical injury.The immunohistochemical results showed that Shh,Smo,Ptch1 and Gli1 staining were mainly located in the endometrium and the u terine gland.No expression of Shh was found in all rats in each group.After curettage,the expression of Smo was very weak,and it was increased gradually in 72 hours after curettage.The expression of Ptch1 was weakly expressed at 24 hours after curettage and increased significantly at 72 hours after curettage.The expression of Gli1 was significantly in creased at 72 hours and 7 days after curettage.The results of real time quantitative PCR showed that Shh mRNA had no expression in the uterus of each time point of each group.Compared with the normal group,the expression of Smo mRNA was 0.7 times (P＜0.05) at 24 hours after curettage,the expression of Ptch1 mRNA was 1.2 times (P＞0.05),and the expression of Gli1 mRNA was 1.6 times (P＜0.05).At 72 hours after curettage,the expression of Smo mRNA was 1.4 times (P＜0.05),the expression of Ptch1 mRNA was 3.3 times (P＜0.01),and the expression of Gli1 mRNA was 9.3 times (P＜0.01).At 7 days after curettage,the expression of Smo mRNA was 1.5 times (P＜0.05),the expression of Ptch1 mRNA was 2.5 times (P＜0.05),and the expression of Gli1 mRNA was 5.7 times (P＜0.01).Conclusion:Ptch1,Smo and Gli1 are all expressed in rats' endometrium after mechanical injury.The expression of Ptch1 mRNA,Smo mRNA and Gli1 mRNA are related to the rat endometrium fibrosis.Hedgehog (Hh) signaling pathway may be involved in the endometrium fibrosis repair after mechanical injury.
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