雷公藤内酯醇对人近端肾小管上皮细胞单核细胞趋化因子合成的影响

TRIPTOLIDE INHIBITS MCP-1 OVERPR ODUCTION IN HUMAN PROXIMAL TUBULA R CELLS INDUCED BY IFNγ AND TNFα COSTIMU LATION

目的 :观察雷公藤内酯醇对炎症因子刺激下人肾小管上皮细胞单核细胞趋化因子 (MCP 1)产生的影响。 方法 :以IFN γ(2 0 0 μg/L)及TNFα(2 0 μg/L)联合刺激人近端小管上皮细胞 ,细胞MCP 1蛋白含量以流式细胞仪检测 ,培养上清中MCP 1的分泌量以双抗体夹心ELISA法测定 ,细胞MCP 1mRNA含量采用半定量RT PCR法进行测定。 结果 :普通培养人近端肾小管上皮细胞及培养上清中有少量MCP 1分泌 ,IFN γ(2 0 0 μg/L)及TNFα(2 0μg/L)联合刺激 2 4h ,人近端小管上皮细胞合成MCP 1明显增加。雷公藤内酯醇可以抑制肾小管上皮细胞MCP 1分泌 ,并使细胞MCP 1mRNA含量明显降低 ,该抑制效应呈剂量依赖性。 结论 :雷公藤内酯醇可以抑制炎症刺激下人近端肾小管上皮细胞合成MCP 1,这可能是雷公藤内酯醇产生其疗效的机制之一。

To inv estigate the effects of triptolide in in hibiting the MCP-1 production in human p roximal tubular cells. METHO DOLOGY A human proximal tubu lar cell line was selected.A combination of IFN-γ(200 μg/L)and TNFα(20 μg/L)was used to stimulate the production of MCP- 1 in tubular cells.Intracellular MCP-1 w as detected by flowcytometry and MCP-1 i n the supernatant was measured by “sandw ich” ELISA method.MCP-1 mRNA expression was detected by semi-quantitative polyme rase chain reaction. RESULTS In unstimulated tubular cel ls,a mild of MCP-1 was detected in cells and in culture supernatants.In the supe rnatants of tubular cells costimulated with IFN-γ(200 μg/L)and TNFα(20 μg/L)for 24 hours,MCP-1 production was significa ntly increased (1120.9±71.9 vs 141.9±1 7.3 ng/L·μg protein -1,P<0.01),a simi lar increase of intrac ellular MCP-1 protein was observed with FA CS analysis.MCP-1 mRNA expression was in creased nearly two folds of that in uns tim ulated cells.Triptolide could dose-depen dently inhibit MCP-1 protein production( 713.1±28.4 vs 1120.9±71.9 ng/L·μg prot ein -1,P<0.01)and mRNA expression sti mulated by inflammatory factors. CONCLUSION Triptolide can inhibit M CP-1 overproduction in human tubular cel ls induced by IFN-γ(200 μg/L)and TNFα(20 μg/L)costimulation.This may be one of t he mechanisms by which triptolide excert s in treatment of renal diseases.