系膜细胞来源的白细胞介素-13对系膜细胞细胞因子基因表达的研究(英文)

Effect of mesangial cell2derived interleukin213 on expression of cytokines synthesized human mesangial cells

摘要:

目的 白细胞介素 1 3(IL 1 3)是新近发现的一种抗炎性细胞因子 ,其在肾小球肾炎中的作用尚不清楚 ,该研究探讨脂多糖 (LPS)对体外培养的人肾小球系膜细胞 (HMC)表达IL 1 3作用以及IL 1 3对HMC促炎性细胞因子、趋化因子和促纤维化因子基因表达的影响。方法 体外培养HMC ,加入不同浓度的LPS和 (或 )IL 1 3后 ,用逆转录 -聚合酶链反应和ELISA检测HMCIL 1 3mRNA表达和细胞培养上清液中IL 1 3蛋白含量 ;应用核酸酶保护法检测HMC肿瘤坏死因子 α(TNF α)、白介素 - 1α(IL 1α)、白介素 - 1 β(IL 1 β)、单核细胞趋化蛋白 1(MCP 1 )、白介素 8(IL 8)、转化生长因子 - β1 (TGF β1 )mRNA的表达。 结果 未予LPS刺激的HMC不表达IL 1 3mRNA和蛋白 ;LPS呈剂量依赖性和时间依赖性诱导HMC表达IL 1 3mRNA和分泌IL 1 3蛋白。HMC受LPS刺激后 1 2h即可表达IL 1 3mRNA ,4 8h达高峰 ,72h仍维持在较高的水平。HMC受LPS刺激后 2 4h ,其培养上清液中检测到IL 1 3蛋白 ,4 8h和 72h进一步增加。外源性IL 1 3呈剂量依赖性地抑制LPS诱导的系膜细胞TNF α ,IL 1α ,IL 1 β ,MCP 1 ,IL 8,TGF β1mRNA的表达。应用抗IL 1 3抗体中和内源性IL 1 3后 ,上述炎症因子表达增强。结论 IL 1 3是HMC自分泌因子。IL 1 3可抑制LPS诱导

Abstract:

Objective This study aims to investigate the interleukin 13 (IL 13) expression in the human mesangial cells (HMC) and its effect on expressions of cytokines synthesized by HMC so as to study the role of IL 13 in the inflammatory process of glomerulonephritis. Methods The HMC were cultured and treated with LPS and/or recombinant human IL 13. The IL 13 mRNA expression and the IL 13 protein level in the cultured HMC were detected by semiquantitative reverse transcription polymerase chain reaction (RT PCR) and enzyme linked immunosorbent assay (ELISA) respectively. The effects of IL 13 on the expressions of proinflammatory cytokines, chemokines and profibrogenic cytokine in the HMC were determined by ribonuclease protection assay (RPA). The cultured HMC without LPS or recombinant human IL 13 stimulation were used as the controls. Results The IL 13 mRNA expression and IL 13 protein were undetected in the controls without stimulation. The IL 13 mRNA expression in the HMC with LPS stimulation was induced as early as 12 hrs after LPS stimulation, reached a peak at 48 hrs and remained high level until 72 hrs. The activation of HMC with LPS resulted in the induction of IL 13 mRNA expression in a dose and time dependent way. The IL 13 protein was induced 24 hrs after LPS stimulation and was further increased with stimulation time. The recombinant IL 13 inhibited TNF α, IL 1α, IL 1β, MCP 1, IL 8 and TGF β1 mRNA expressions were induced by LPS in a dose dependent way. The expressions of TNF α, IL 1α, IL 1β, MCP 1, IL 8, and TGF β1 mRNA were increased after endogenously produced IL 13 was neutralized with anti IL 13 mAb. Conclusions LPS can induce the IL 13 expression in HMC. The mesangial cell derived IL 13 can inhibit the production of proinflammatory cytokines, chemokines, and profibrogenic cytokine synthesized by HMC.