晚期糖化终产物受体、核因子-κB 双基因反义RNA抑制糖化终产物刺激的炎症因子表达

中国临床药理学与治疗学 ›› 2006, Vol. 11 ›› Issue (7): 784-788.

晚期糖化终产物受体、核因子-κB 双基因反义RNA抑制糖化终产物刺激的炎症因子表达

游捷^1,2, 赵蓉², 刘礼斌¹, 林建银²

¹福建医科大学附属协和医院省内分泌研究所, 福州 350001, 福建;
²福建医科大学分子医学研究中心, 福州 350004, 福建

收稿日期:2006-03-14 修回日期:2006-06-10 出版日期:2006-07-26 发布日期:2020-10-30
作者简介:游捷, 女, 博士生, 副主任医师, 研究方向:糖尿病慢性并发症。Tel:0591-83357896-8452 E-mail:youjie987@sina.com.cn
基金资助:
福建省科技开发计划项目(No2003D09); 福建省卫生厅青年科研基金(No2004-1-1)

Inhibitory effect of receptor of advanced glycation endproducts, nuclear factor-κB double gene antisense RNA on productions of inflammatory factor treated by advanced glycation endproducts

YOU Jie^1,2, ZHAO Rong², LIU Li-bin¹, LIN Jian-yin¹

¹Department of Endocrinology, Union Hospital , Fujian Medical University, Fuzhou 350001 , Fujian , China;
²Molecular Medical Centre, Fujian Medical University , Fuzhou 350004, Fujian, China

Received:2006-03-14 Revised:2006-06-10 Online:2006-07-26 Published:2020-10-30

摘要/Abstract

摘要： 目的观察晚期糖化终产物受体(receptor of advanced glycation endproducts, RAGE ) 、核因子-κB(NF-κB) 双基因反义RNA 对晚期糖化终产物(advanced glycation endproducts, AGEs) 刺激ECV304 细胞分泌炎症因子的影响。方法酶联免疫吸附测定(enzyme linked immunosorbent assay, ELISA) 法观察AGEs 对ECV304 细胞分泌TNF-α和IL-6 的影响, 应用脂质体将RAGE 、NF-κB 单/双基因反义RNA 转染ECV304, 流式细胞仪、筛选低表达RAGE 、NF-κB 的细胞株, 观察RAGE 、NF-κB 双基因反义RNA 对AGEs刺激ECV304 细胞分泌TNF-α和IL-6 的影响。结果 AGEs 可引起ECV304 细胞分泌TNF-α和IL-6 增加, 诱导作用具有时间和剂量依赖规律。稳定转染筛选出低表达RAGE 、NF-κB 的细胞株, 在AGEs100 mg·L^-1 诱导下双基因转染细胞ECV-asRAGEasP65克隆中RAGE 、NF-κBp65 表达抑制率分别为(62.2±8.7) %及(37.2±7.1) %。AGEs 100 mg·L^-1刺激下ECV-asRAGE-asP65 克隆分泌TNF-α、IL-6 较空载体转染细胞、单基因转染细胞减少更明显(P<0.01) 。结论 RAGE 、NF-κB 双基因反义RNA 可抑制AGEs 刺激的ECV304 细胞的TNF-α和IL-6 释放。

关键词: RNA, 糖基化终产物, 受体, 核因子-κB, 基因转染, 炎症因子

Abstract: AIM: To observe the effects of receptor of advanced glycation endproducts(RAGE) , NF-κB double gene antisense RNA on the productions of TNF-αand IL-6 treated by advanced glycation endproducts (AGEs ).METHODS: The levels of TNF-αand IL-6 in the supernatants were measured by enzyme linked immunosorbent assay (ELISA) in ECV304 cells treated by AGEs.The RAGE, NF-κB single double gene antisense RNA were transfected into ECV304 cell.The expressions of RAGE and NF-κB were detected by flow cytometry and RT-PCR.In different clone cells, the effects of antisense RNA on the productions of TNF-α、IL-6 were determined by ELISA.RESULTS: AGEs, instead of human serum albumin (HSA) , stimulated ECV304 cell to produce TNF-α and IL-6 with a time-and dose-dependent manner.The RAGE, NF-κB single double gene antisense RNA were transfected into ECV304 cell.Induced by AGEs, the expressions of RAGE and NF-κB in double gene cotransfected cell were inhibited by (62.2 ±8.7) % and (37.2 ±7.1) %, respectively.Induced by AGEs, the amount of TNF-α、IL-6 in the medium were lower in single gene transfected cells ECV-asRAGE, ECV-asP65 than ECVVector(P <0.05).The amount of TNF-α、IL-6 in the double gene transfected cells ECV-asRAGE-asP65 were lower than ECV-Vector and ECV-asRAGE, ECV-asP65(P<0.05).CONCLUSION: The production of TNF-α and IL-6 is increased in ECV304 cell treated by AGEs.RAGE, NF-κB double gene antisense RNA can inhibited the production of inflammatory factor treated by AGEs.

Key words: antisense RNA, advanced glycation endproducts, receptor, nuclear factor-κB, gene transfection, inflammatory factor

中图分类号:

R965.2
R966

游捷, 赵蓉, 刘礼斌, 林建银. 晚期糖化终产物受体、核因子-κB 双基因反义RNA抑制糖化终产物刺激的炎症因子表达[J]. 中国临床药理学与治疗学, 2006, 11(7): 784-788.

YOU Jie, ZHAO Rong, LIU Li-bin, LIN Jian-yin. Inhibitory effect of receptor of advanced glycation endproducts, nuclear factor-κB double gene antisense RNA on productions of inflammatory factor treated by advanced glycation endproducts[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2006, 11(7): 784-788.

参考文献

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