15d-PGJ_2对肾小管上皮细胞CD40及RANTES表达的影响  

Effect of 15d-PGJ_2 on the expression of CD40 and RANTES induced by IFN-γ and TNF-α on renal tubular epithelial cells(HK-2)

作　　者：张亚杰[1] 阳晓[1] 张云芳[1] 陈伟英[1] 孔庆瑜[1] 董秀清[1] 李晓艳[1] 余学清[1] 

机构地区：[1]中山大学附属第一医院肾内科教育部卫生部肾脏病临床研究重点实验室,广州510080

出　　处：《中国免疫学杂志》2006年第10期889-894,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金(30271668;30572290);教育部博士点专项基金(2002055805);广东省重点攻关项目(2003C30202)资助

摘　　要：目的:观察过氧化物酶体增殖物激活受体γ(PPARγ)激动剂15d-PGJ2对干扰素-γ(IFN-γ)和肿瘤坏死因子α(TNF-α)诱导的人近端肾小管上皮细胞(HK-2)CD40和RANTES表达的影响。方法:体外培养人近端肾小管上皮细胞株,分组如下:(1)正常对照组;(2)IFN-γ(50μg/ml)组;(3)TNF-α(10ng/ml)组;(4)IFN-γ(50μg/ml)+TNF-α(10ng/ml)组;(5)IFN-γ(50μg/ml)+TNF-α(10ng/ml)分别加1、3、5μmol/L15d-PGJ2组;(6)IFN-γ+TNF-α刺激+15d-PGJ2(5μmol/L)+GW9662(PPARγ特异拮抗剂)1μmol/L组。GW9662和15d-PGJ2分别在IFN-γ和TNF-α刺激前3和2小时加入。分别采用RT-PCR、流式细胞仪(FACS)和酶联免疫法(ELISA)检测CD40和RANTES基因和蛋白的表达水平。结果:正常HK-2细胞中,CD40、RANTES有基础水平表达。IFN-γ能显著上调HK-2细胞CD40蛋白的表达;TNF-α单独刺激对CD40表达无显著影响,但能增强IFN-γ的刺激效应。IFN-γ+TNF-α显著增加HK-2细胞RANTES的表达和分泌。15d-PGJ2呈剂量依赖式在基因和蛋白水平显著抑制IFN-γ+TNF-α诱导的CD40和RANTES的表达。加入PPARγ特异拮抗剂GW9662后,能够部分逆转15d-PGJ2对CD40和RANTES表达的抑制效应,但并不能完全阻断其作用。结论:15d-PGJ2部分通过PPARγ介导的信号途径参与抑制IFN-γ+TNF-α诱导的人近端肾小管上皮细胞CD40和RANTES的表达,从而在肾脏局部发挥免疫调节和抗炎作用。Objective ：To observe the effect of PPARγ, agonists 15d-PGJ2 on the expression of CD40 and RANTES in cultured proximal tubule cells （HK-2） induced by IFN-γ, and TNF-α. Methods：HK-2 cells were cultured in DMEM/F12 with 10% fetal bovine serum in vitro and were divided into following groups： IFN-γ（50 μg/ml） group, TNF-α（ 10 ng/ml） group,IFN-T（50 μg/ml） ＋TNF- α（10 ng/ml） group, IFN-γ（50 μg/ml） ＋TNF-α（10 ng/ml） with 1,3,5 μmol/L 15d-PGJ2 groups and IFN-γ（50μg/ml） ＋TNF-α （10 ng/ml） with 15d-PGJ2（5 μmol/L） and GW9662（ 1 μmol/L） group. The expression of CD40 mRNA and protein were measured by RT-PCR and flow cytometry（FACS） respectively. The expression of RANTES mRNA and protein were measured by RT-PCR and ELISA respectively. Results： HK-2 cells expressed low levels of CD40 and RANTES. Activation of HK-2 cells by IFN-γ, resulted in an enhanced expression of CD40; the present of TNF-α alone did not result in significant expression of CD40, but simultaneous activation of HK-2 cells by TNF-α and IFN-γ, result in strong synergistic effect on the expression of CD40 Combinations of TNF-α together with IFN-γ, resulted in up-regulated RANTES mRNA and strong induction of RANTES production. The PPARγ, agonists 15-deoxy-12,14- prostaglandin J2（ 15d-PGJ2 ） significantly blocked CD40 and RANTES expression in HK-2 cell line induced by IFN-γ, plus TNF-α. GW9662 partly abrogated the inhibition of CD40 and RANTES induced by 15d-PGJ2. Conclusion： 15d-PGJ2 decreased CD40 and RANTES expression induced with IFN-γ, plus TNF-α in HK-2 cells,and these immtme modulation and anti-inflammation effects were partially mediated by PPAR-dependent pathways.

关 键 词：过氧化物酶体增殖物激活受体Γ 肾小管上皮细胞 CD40 RANTES 

分 类 号：R392.11[医药卫生—免疫学]