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目的探讨脂多糖(LPS)引起人腹膜间皮细胞(HPMC)损伤中分泌MCP-1以及LPS、P38MAPK、MCP-1三者之间可能存在的关系。方法体外培养永生化人腹膜间皮细胞(HPMC),随机分为正常对照组、LPS作用24 h组、LPS作用48 h组、特异性P38MAPK阻断剂SB203580+LPS作用24 h组、SB203580+LPS作用48 h组;Western blot法检测各组MCP-1和磷酸化P38MAPK蛋白表达水平;Real-time PCR法检测各组MCP-1 mRNA表达水平。结果 1.10 mg/L LPS刺激使HPMC的MCP-1 mRNA和蛋白质表达均较正常对照组增加(P<0.05)。10 mg/L LPS作用48 h后MCP-1 mRNA和蛋白质表达水平均高于24 h组(P<0.05);Western blot结果显示,与正常组对比,10 mg/L LPS作用使磷酸化P38MAPK蛋白水平明显升高(P<0.05)。10 mg/L LPS作用48 h与作用24 h对比升高不明显(P>0.05);经5μmol/L SB203580预处理30 min后再予以LPS刺激与单纯LPS刺激相比较,MCP-1蛋白质和mRNA均明显降低(P<0.05);SB203580预处理后再予以LPS分别刺激24 h和48 h 2组相比较,MCP-1蛋白及mRNA水平差异不明显(P<0.05)。结论 LPS通过磷酸化P38MAPK,导致MCP-1表达水平升高,诱发腹膜间皮细胞的损伤。
Objective To investigate the possible relationship between the secretion of MCP-1, LPS, P38MAPK and MCP-1 in human peritoneal mesothelial cells (HPMC) induced by lipopolysaccharide (LPS). Methods Immortalized human peritoneal mesothelial cells (HPMC) were cultured in vitro and randomly divided into normal control group, LPS group for 24 hours, LPS group for 48 hours and SB203580 + LPS group for 24 hours. SB203580 + LPS The expression of MCP-1 and phosphorylated P38MAPK protein in each group was detected by Western blot. The expression of MCP-1 mRNA in each group was detected by Real-time PCR. Results The mRNA and protein expressions of MCP-1 in HPMC increased with stimulation with LPS at 1.10 mg / L (P <0.05). The expression of MCP-1 mRNA and protein were significantly increased after treatment with 10 mg / L LPS for 48 h (P <0.05). Compared with the normal group, the expression of MCP-1 mRNA and protein was up-regulated by 10 mg / L LPS The level was significantly higher (P <0.05). Compared with LPS stimulation, the protein and mRNA expression of MCP-1 were significantly increased after pretreatment with 5μmol / L SB203580 for 30min (P <0.05) (P <0.05). Compared with the pretreatment with SB203580 for 24 h and 48 h, the MCP-1 protein and mRNA levels were not significantly different (P <0.05). Conclusion LPS phosphorylates P38MAPK, which leads to the increase of MCP-1 expression and induces the injury of peritoneal mesothelial cells.