15d-PGJ2对肾小管上皮细胞CD40及RANTES表达的影响
Effect of 15d-PGJ2 on the expression of CD40 and RANTES induced by IFNγ and TNFα on renal tubular epithelial cells
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γ-干扰素（IFNγ）和肿瘤坏死因子α（TNFα）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）IFNγ（50μg/ml）组；（3）TNFα(10ng/ml)组；（4）IFNγ（50μg/ml）＋TNFα(10ng/ml)组；(5)IFNγ（50μg/ml）＋TNFα(10ng/ml)分别加1、3、5μmol/L 15d-PGJ2组；(6)IFNγ+TNFα刺激＋15d-PGJ2 (5μmol/L)＋GW9662（PPARγ特异拮抗剂）1μmol/L组。GW9662和15d-PGJ2分别在IFNγ和TNFα刺激前3h 和2h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。IFNγ能显著上调HK-2细胞CD40蛋白的表达；TNFα单独刺激对CD40表达无显著影响，但能增强IFNγ的刺激效应。IFNγ＋TNFα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IFNγ＋TNFα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IFNγ+TNFα诱导的人近端肾小管上皮细胞CD40和RANTES的表达，从而发挥免疫调节和抗炎效应。
o observe the effect of PPARγagonists 15d-PGJ2 on the expression of CD40 and RANTES in cultured proximal tubule cells (HK-2) induced by IFNγ and TNFα. Methods HK-2 cells were cultured in DMEM/F12 with 10% fetal bovine serum.in vitro and were divided into following groups: IFNγ（50μg/ml）group, TNFα(10ng/ml)group，IFNγ（50μg/ml）＋TNFα(10ng/ml) group, IFNγ（50μg/ml）＋TNFα(10ng/ml) with 1,3,5μmol/L 15d-PGJ2 groups and IFNγ（50μg/ml）＋TNFα(10ng/ml) with 15d-PGJ2 (5μmol/L) and GW9662（1μmol/L） group. The expression of CD40mRNA 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 IFNγ resulted in an enhanced expression of CD40; the present of TNFα alone did not result in significant expression of CD40, but simultaneous activation of HK-2 cells by TNFα and IFNγ result in strong synergistic effect on the expression of CD40 Combinations of TNFα together with IFNγ 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 IFNγplus TNFα. GW9662 partly abrogated the inhibition of CD40 and RANTES induced by 15d-PGJ2. Conclusion 15d-PGJ2 decreased CD40 and RANTES expression induced with IFNγplus TNFαin HK-2 cells，and these immune modulation and anti -inflammation effects were partially mediated by PPAR-dependent pathways.
张亚杰、张云芳、阳晓、孔庆瑜、李晓艳、陈伟英、余学清、董秀清
基础医学生物化学生理学
过氧化物酶体增殖物激活受体γ 肾小管上皮细胞 CD40 RANTES
PPARγRenal tubular epithelial cells CD40 RANTES
张亚杰,张云芳,阳晓,孔庆瑜,李晓艳,陈伟英,余学清,董秀清.15d-PGJ2对肾小管上皮细胞CD40及RANTES表达的影响[EB/OL].(2005-12-12)[2025-08-16].http://www.paper.edu.cn/releasepaper/content/200512-253.点此复制
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