环孢素对同种异基因大鼠子宫移植急性排斥反应的影响

The immunosuppressive effect of cyclosporine(CsA) in allograft uterus transplantation in rats

目的:探讨免疫抑制剂环孢素对同种异基因大鼠子宫移植排斥反应的影响及其机制。方法:以SD大鼠为供体,Wistar大鼠为受体,建立同种异基因大鼠子宫移植排斥模型。移植后第一天起使用环孢素,每天每公斤体重10mg肌肉注射;对照组以等量生理盐水替代。根据受体大鼠应用环孢素的时间,分为3组,即移植后7天(POD-7)、14天(POD-14)及28天(POD-28),每组10对。对照组仅移植后7天组10对。观察移植子宫的排斥反应,应用Real-time PCR方法检测移植物内细胞因子IL-2,IFN-γ,TGF-β1,IL-10在各组的动态变化,并用免疫组化方法检测移植子宫雌激素受体的表达。结果:(1)实验组移植子宫在观察期内未见明显排斥反应,对照组则在7天时出现严重的排斥反应,移植子宫明显坏死;(2)实验组移植物内IL-2,IFN-γ表达量明显少于对照组(P<0.05),TGF-β1表达亦下调(P<0.05),而POD-14组与POD-28组IL-10表达上调(P<0.05);(3)对照组ER表达缺失,而实验组ER表达强度与用药时间呈负相关(P<0.01)。结论:环孢素能有效地抑制子宫移植的急性排斥反应。主要通过调节移植子宫的Th1/Th2细胞因子平衡向Th2漂移起作用。长期应用环孢素对大鼠移植子宫的内分泌功能有一定的影响。

Objective：To explore the immunosuppressive effect of cyclosporine （CsA） in allograft uterus transplantation in rats and its mechanisms. Methods： Heterotopic uterine transplantations were performed on Wistar rats and grafts were harvested on postoperation days 7,14,28. They were divided into two groups ： control group ： CsA-untreated group （ n = 10） ; experimental group：dosage of CsA-treated group （10mg/kg per day）. The grafts were examined under microscopy by HE stain. The intragraft mRNA expression levels of IL-2 ,IFN-γ,TGF-β1, IL-10 were detected by real-time quantitative PCR. And the estrogen receptor （ER） proteins were detected by Immuohistochemical analysis. Results： （ 1 ） The histopathological examination showed that the grafts displayed edema,black and stiffness on 7 days after uterine transplantation in the control group;but in the experimental group, the survival grafted uterus had no signs of necrosis and demonstrated normal morphology by histopathological examination ; （2） The expression levels of IL-2, IFN-γ, were significantly lower in the experimental group as compared with control group （ P 〈 0.05 ）, and TGF-β1 also lower in the experimental group （ P 〈 0.05 ）. However,The IL-10 expression levels were increased in the POD-14 and POD-28 group;（3） The protein expression of rat ER gene was not detected in control group. There were positive expression in the experimental group, and the expression level had a negative correlation with the dose of CsA（P 〈 0.01 ）. Conclusions： CsA has significant therapeutic effect on the allograft rejection of uterine transplantation in rats. Maybe, It works by inducing the balance of Thl/Th2 cytokine shifting from Thl to Th2. The long-term use of CsA has impact on the reproductive endocrine function in rats.