[1]秦琴,石韵洁,赵青青,等.CD25siRNA基因转染可提高大鼠穿透性角膜移植片的存活率[J].陆军军医大学学报(原第三军医大学学报),2015,37(15):1543-1549.
 Qin Qin,Shi Yunjie,Zhao Qingqing,et al.CD25siRNA transfection in corneal graft improves its survival by up-regulating IL-10 in rats after penetrating keratoplasty[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(15):1543-1549.
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CD25siRNA基因转染可提高大鼠穿透性角膜移植片的存活率(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
37卷
期数:
2015年第15期
页码:
1543-1549
栏目:
论著
出版日期:
2015-08-15

文章信息/Info

Title:
CD25siRNA transfection in corneal graft improves its survival by up-regulating IL-10 in rats after penetrating keratoplasty
作者:
秦琴石韵洁赵青青陈元吴静赵敏
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
Author(s):
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

关键词:
CD25siRNA移植排斥反应纳米转染试剂IL-10FOXP3
Keywords:
CD25siRNA graft rejection nanoparticle transfection reagents IL-10 FOXP3
分类号:
R392.11; R394.3; R779.65
文献标志码:
A
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50  μL EntransterTM-反义CD25siRNA和50 μL EntransterTM-CD25siRNA 点眼纳米转染试剂,50  μL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。

Abstract:

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 μL EntransterTM-control CD25siRNA and 50  μL EntransterTM-CD25siRNA were formulated as nanoparticle transfection reagents, and 50  μL 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, EntransterTM-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 alleviate infiltration of inflammation cells and reduce neovascularization. Meanwhile, lower CD25 expression and higher IL-10 expression were detected in the EntransterTM-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.

更新日期/Last Update: 2015-08-19