CD25siRNA基因转染可提高大鼠穿透性角膜移植片的存活率

发布时间：2018-06-08 01:24

本文选题：CDsiRNA + 移植排斥反应　；参考：《第三军医大学学报》2015年15期

【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(P0.05);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(P0.05),但IL-10表达均高于其余2组(P0.05),FOXP3表达在3组间无统计学意义(P0.05)。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。
[Abstract]:Objective to investigate the effect of CD25siRNA gene transfection on penetrating corneal allograft rejection in rats and the changes of IL-10 and FOXP3 levels in grafts after transplantation. Methods 50 渭 L EntransterTM- antisense CD25siRNA and 50 渭 L EntransterTMCD25 siRNA were prepared as control. The penetrating corneal transplantation model of Wistar SD rats was established. The right eye was used as the operative eye. The model was divided into control group (group A) and EntransterTM-CD25siRNA transfection group (group C, 28 rats in each group) by a simple random sampling method. The rats were divided into two groups: control group A (n = 28) and EntransterTM-CD25siRNA transfected group (n = 28). The grafts were graded daily and HE staining was performed on 21 days after operation. Western blot and qRT-PCR were detected by immunohistochemistry. Results the survival rate of corneal grafts in CD25siRNA transfected group was significantly better than that in the other two groups (P 0.05). The corneal graft neovascularization and inflammatory cell infiltration were decreased in CD25siRNA transfection group as compared with the other two groups, meanwhile, the survival rate of corneal graft in CD25siRNA transfection group was significantly higher than that in the other two groups. The expression of CD25 in CD25siRNA transfection group was lower than that in the other two groups, but the expression of IL-10 was higher than that of the other two groups. There was no significant difference between the three groups in the expression of FOXP3. Conclusion CD25siRNA transfection inhibits inflammatory response by upregulating the expression of anti-inflammatory cytokine IL-10, thus prolonging the survival time of corneal grafts and increasing the survival rate of grafts.
【作者单位】：重庆医科大学附属第一医院眼科重庆市眼科学重点实验室;
【基金】：国家自然科学基金面上项目(81170822)~~
【分类号】：R779.6

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