siRNA干预树突状细胞CD40表达对大鼠炎症性肠病的治疗作用

doi:10.13481/j.1671-587x.20150111

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (01): 54-59.doi: 10.13481/j.1671-587x.20150111

siRNA干预树突状细胞CD40表达对大鼠炎症性肠病的治疗作用

王丽英¹, 杨勇², 李占东³, 李东复¹, 刘力宾¹, 王敏¹

1. 吉林大学第二医院消化内科, 吉林长春 130041;
2. 解放军第208医院消化内科, 吉林长春 130062;
3. 吉林工程技术师范学院食品工程学院生物工程教研室, 吉林长春 130052

收稿日期:2014-10-15 发布日期:2015-01-30
通讯作者: 王丽英,副教授,硕士研究生导师(Tel:0431-88796807,E-mail:wly2588@163.com) E-mail:wly2588@163.com
作者简介:王丽英(1965-),女,山东省高密市人,副教授,医学博士,博士后,主要从事消化系统疾病的诊断及治疗方面的研究。
基金资助:
吉林省科技厅自然科学基金资助课题(201115086)

Therapentic effect of siRNA inhibiting CD40 expression in dendritic cells on inflammatory bowel disease in rats

WANG Liying¹, YANG Yong², LI Zhandong³, LI Dongfu¹, LIU Libin¹, WANG Min¹

1. Department of Gastroenterology, Second Hospital, Jilin University, Changchun 130041, China;
2. Department of Gastroenterology, No.208 of PLA, Changchun 130062, China;
3. Department of Biological Engineering, College of Food Engineering, Jilin Teachers' Institude of Engineering and Technology, Changchun 130052, China

Received:2014-10-15 Published:2015-01-30

摘要/Abstract

摘要：

目的: 应用小干扰RNA(siRNA)沉默树突状细胞(DCs)CD40分子的表达,观察其对大鼠炎症性肠病(IBD)的影响。方法: 设计并合成高效的siRNA载体及真核表达载体pcDNA3-CD40,分别转染入大鼠DCs细胞。采用PCR和Western blotting法检测DCs中CD40 mRNA及蛋白表达水平。18只Wistar大鼠采用2,4,6-三硝基苯磺酸(TNBS)建立IBD大鼠模型,分为对照组、干预组及过表达组。对照组大鼠给予生理盐水腹腔注射;干预组大鼠给予含有干涉表达载体pSilencer 4.1-CMV neo-CD40的DCs腹腔注射;过表达组大鼠给予含有pcDNA3-CD40的DCs腹腔注射。15 d后观察各组大鼠的腹泻、黏液脓血便等临床症状。结果: 成功构建RNA干涉载体并转染DCs,PCR及Western blotting法检测DCs中CD40 mRNA及蛋白表达水平均下调。转染后的DCs作用于各组大鼠,与对照组比较,干预组大鼠的腹泻、黏液脓血便症状得到明显改善;过表达组大鼠腹泻、黏液脓血便情况加重,并有2只大鼠死亡。结论: siRNA干预DCs 中CD40分子的表达对大鼠IBD具有治疗作用。

关键词: 炎症性肠病, 树突状细胞, 小干扰RNA, CD40

Abstract:

Objective To use small interfering RNA (siRNA) to silence the expression of CD40 molecule in dendritic cells (DCs), and to observe its effect on inflammatory bowel disease (IBD) in the rats. Methods A highly efficient siRNA carrier and eukaryotic expression vector pcDNA3-CD40 were designed and synthesized, and they were transfected into DCs of the rats respectively to build the inflammatory bowel disease (IBD) models in the rats.18 Wistar rats were divided into control group, intervention group and overexpression group .All rats models were built using trinitro-benzene -sulfonic acid(TNBS).After successful construction, the rats in control group were injected intraperitoneally with normal saline, the rats in intervention group were injected intraperitoneally with interference expression vector, and the rats in overexpression group were injected intraperitoneally with pcDNA3-CD40.The clinical symptoms such as diarrhea and mucopurulent bloody stool were observed after 15 d. Results The interference expression vector was constructed successfully.The expression levels of CD40 mRNA and protein in DCs were both reduced detected by PCR and Western blotting method.The transfected DCs were successfully acted on model rats.The clinical symptoms such as diarrhea, mucus purulent blood in intervention group were improved significantlycompared with control group.The symptoms such as diarrhea and mucopurulent bloody stool were deteriorated in overexpression group, of which two rats were dead. Conclusion The intervened expression of CD40 in DCs with siRNA has a therapeutic effect on IBD in rats.

Key words: inflammatory bowel disease, dendritic cells, small interfering RNA, CD40

中图分类号:

R-332

王丽英, 杨勇, 李占东, 李东复, 刘力宾, 王敏. siRNA干预树突状细胞CD40表达对大鼠炎症性肠病的治疗作用[J]. 吉林大学学报(医学版), 2015, 41(01): 54-59.

WANG Liying, YANG Yong, LI Zhandong, LI Dongfu, LIU Libin, WANG Min. Therapentic effect of siRNA inhibiting CD40 expression in dendritic cells on inflammatory bowel disease in rats[J]. Journal of Jilin University Medicine Edition, 2015, 41(01): 54-59.

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siRNA干预树突状细胞CD40表达对大鼠炎症性肠病的治疗作用

Therapentic effect of siRNA inhibiting CD40 expression in dendritic cells on inflammatory bowel disease in rats

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