摘要
目的探讨CD25siRNA基因转染对大鼠穿透性角膜移植排斥反应的作用,及术后植片中IL-10和FOXP3的含量变化。方法配制50μL EntransterTM-反义CD25siRNA和50μL EntransterTMCD25siRNA点眼纳米转染试剂,50μL生理盐水为对照。建立Wistar-SD大鼠穿透性角膜移植模型,右眼为术眼,采用单纯随机抽样方法,分为对照组(A组)、EntransterTM-反义CD25siRNA转染组(B组)和EntransterTM-CD25siRNA转染组(C组),每组28只。术后每日对植片行临床评分并分别于3、7、14、21 d行HE染色、免疫组化、Western blot和qRT-PCR检测。结果角膜植片生存曲线显示CD25siRNA转染组角膜植片的生存率显著优于其余2组(P<0.05);HE染色显示CD25siRNA转染组较其余2组,角膜植片新生血管减少,炎症细胞渗入减少;同时,术后各时间点CD25siRNA转染组中CD25表达均低于其余2组(P<0.05),但IL-10表达均高于其余2组(P<0.05),FOXP3表达在3组间无统计学意义(P>0.05)。结论CD25siRNA转染通过上调抗炎性细胞因子IL-10表达,抑制炎症反应,进而延长角膜植片生存时间,提高植片生存率。
Objective To determine the effects of CD25siRNA transfection in corneal graft on the transplantation rejection in rats undergoing penetrating keratoplasty,and on the expression of IL-10 and FOXP3 in the graft.Methods Fifty I^L EntransterTM-control CD25siRNA and 50 p^L EntransterTM-CD25siRNA were formulated as nanoparticle transfection reagents,and 50 txL normal saline was defined as control.Orthotopic corneal transplantation was performed only on the right eyes of Wistar SD rats.Corneal recipients were divided into the control group,EntransterWM-control CD25siRNA group and EntransterTM-cD25siRNA group,with 28 rats in each group.After surgery,the recipient eyes were examined daily for 21 d,and HE staining,immunohistochemistry,Western blotting and qRT-PCR were performed on 3,7,14 and 21 d postoperatively.Results The survival curves indicated that CD25siRNA treatment could significantly prolong graft survival time compared to the other 2 groups(P〈0.05).HE staining showed that CD25siRNA treatment could aUeviate infiltration of inflammation cells and reduce neovascularization.Meanwhile,lower CD25 expression and higher IL-10 expression were detected in the EntransterYM-CD25siRNA group compared with the other 2 groups(P〈0.05),and FOXP3 expression exhibited no significantly difference among the groups(P〉0.05).Conclusion CD25siRNA transfection significantly prolongs the median survival time of corneal allografts and increases the graft survival rate via up-regulating anti-inflammatory molecule IL-10 and inhibiting tissue inflammatory response.
作者
秦琴
石韵洁
赵青青
陈元
吴静
赵敏
Qin Qin;Shi Yunjie;Zhao Qingqing;Chen Yuan;Wu Jing;Zhao Min(Department of Ophthalmology,Chongqing Key Laboratory of Ophthalmology,the First Affiliated Hospital of Chongqing Medical University,Chongqing,400016,China)
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2015年第15期1543-1549,共7页
Journal of Third Military Medical University
基金
国家自然科学基金面上项目(81170822)
