›› 2010, Vol. 30 ›› Issue (5): 535-.

• 论著(基础研究) • 上一篇    下一篇

CTGF反义寡核苷酸对人肾小管上皮细胞转分化的影响

廖晓莉1, 陈 超1,2, 陈 涛1,3, 严玉霞1, 林春兰1, 蒋建伟1   

  1. 1.暨南大学 医学院生物化学教研室, 广州 510632;2.河南省焦作市人民医院普外科, 焦作 454002;3.广东省结核病防治研究所, 广州 510632
  • 出版日期:2010-05-25 发布日期:2010-05-28
  • 通讯作者: 蒋建伟, 电子信箱: jjw703@163.com。
  • 作者简介:廖晓莉(1984—), 女, 硕士生;电子信箱: 719958217@qq.com。
  • 基金资助:

    广东省医学科研基金(A2009348)

Effects of CTGF antisense oligodeoxynucleotide on transition of human kidney epithelial cells

LIAO Xiao-li1, CHEN Chao1,2, CHEN Tao1,3, YAN Yu-xia1, LIN Chun-lan1, JIANG Jian-wei1   

  1. 1.Department of Biochemistry, Jinan University, Guangzhou 510632, China;2.Department of General Surgery, Jiaozuo People's Hospital, Jiaozuo 454002, China;3.Guangdong Institute of Tuberculosis Prevention and Treatment, Guangzhou 510632, China
  • Online:2010-05-25 Published:2010-05-28
  • Supported by:

    Medical Scientific Research Foundation of Guangdong Province, A2009348

摘要:

目的 研究结缔组织生长因子(CTGF)反义寡核苷酸(ASODN)对人肾小管上皮细胞(HKC)转分化的影响。方法 用巨噬细胞趋化因子-1(MCP-1)和马兜铃酸Ⅰ(AAⅠ)联合诱导HKC转分化模型。实验设空白对照组、模型对照组、PEI组、CTGF ASODN 10 ng/mL组和CTGFASODN 100 ng/mL组,分别采用WST-8法、流式细胞术、RT-PCR法、细胞免疫组织化学法检测不同浓度的CTGF ASODN对HKC转分化模型细胞增殖和细胞周期的影响,以及α-SMA mRNA表达和α-SMA、TGF-β1、Fibronectin(FN)蛋白表达的变化。结果 100 ng/mL的CTGF ASODN对MCP-1和AAⅠ联合诱导HKC转分化细胞具有促进细胞增殖的作用,并可减少实验模型导致的细胞S期阻滞,降低α-SMA mRNA表达,减少α-SMA、TGF-β1和FN蛋白的表达。结论 CTGF ASODN能够抑制MCP-1和AAⅠ导致的人HKC转分化。

关键词: 结缔组织生长因子, 反义寡核苷酸, 上皮—间质细胞转分化, 肾小管上皮细胞

Abstract:

Objective To investigate the effects of antisense oligodeoxynucleotide (ASODN) targeting connective tissue growth factor (CTGF) on transition of human kidney epithelial cells (HKC). Methods Transition models of HKC were induced by monocyte chemotactic protein-1(MCP-1) and aristolochic acid Ⅰ(AAⅠ). HKC were randomly divided into blank control group, model control group, PEI group, CTGF ASODN 10 ng/mL group and CTGF ASODN 100 ng/mL group. The effects of CTGF ASODN on proliferation and cell cycle of transition models of HKC were evaluated by WST-8 method and flow cytometry, respectively. The expression of α-smooth muscle actin (α-SMA) mRNA and α-SMA, Fibronectin (FN) and TGF-β1 protein of transition models of HKC were detected by RT-PCR and immunohistochemistry, respectively. Results 100 ng/mL CTGF ASODN reversed the proliferation inhibition, reduced S phase arrest, decreased the expression of α-SMA mRNA and α-SMA, FN and TGF-β1 protein of HKC induced by MCP-1 and AAⅠ. Conclusion CTGF ASODN can inhibit the transition of HKC induced by MCP-1 and AAⅠ.

Key words: connective tissue growth factor, antisense oligodeoxynucleotide, epithelial-to-mesenchymal transition, human kidney epithelial cell