脂多糖对大鼠腹膜间皮细胞白细胞介素15、6及丙二醛表达的影响

The effects of lipopolysaccharide on the expression of inflammatory factors interleukin 15 and interleukin 6 and malondialdehyde in rat peritoneal mesothelial cells

目的研究脂多糖(lipopolysaccharide,LPS)对大鼠腹膜间皮细胞(rat peritoneal me-sothelial cells,RPMC)白细胞介素15(interleukin-15,IL-15)、IL-6及丙二醛(malondialdehyde,MDA)的影响。方法胰蛋白酶法行RPMC的原代培养和传代,经鉴定第3代用于试验。分组:①正常对照组;②LPS组:不同浓度LPS组(1、10、100mg/L)分别作用6h和12h;③不同时间组:10mg/L LPS分别作用于RPMC3、6、12、24h。实时定量PCR法测IL-15mRNA的表达;ELISA法检测细胞培养上清液中IL-15和IL-6的表达;硫代巴比妥法测MDA。结果 LPS可刺激RPMC的IL-15mRNA及蛋白的表达增高且在12h内呈时间剂量依赖性(P<0.05);LPS诱导RPMC IL-6的蛋白表达增高(P<0.05);LPS诱导RPMC MDA的含量表达增高且呈浓度依赖(P<0.05)。结论 IL-15作为炎症因子参与了腹膜透析相关性腹膜炎的病理过程。LPS可上调RPMC IL-15、IL-6和MDA的表达,导致腹膜炎症及氧化应激。

Objective To observe the effects of lipopolysaccharide （LPS） on the expression of inflammatory factors interleukin 15 （IL-15） and interleukin 6 （IL-6） and malondialdehyde （MDA） in rat peritoneal mesothelial cells （RPMCs）. Methods RPMCs were isolated and cultured using trypsin, and then identified. RPMCs at passage 3 were used in the experiment. The cells were divided into （i） normal control group; （ii） LPS dose/time group, LPS 1mg/L, 10mg/L or 100mg/L treated for 6 hours or 12 hours; （iii） LPS time group, LPS 10mg/L treated for 3, 6, 12 and 24 hours. IL-15 mRNA was measured by real-time RT-PCR. IL-15 and IL-6 were assayed by ELISA. MDA were determined by thiobarbituric acid method. Results IL-6, IL-15 and its mRNAs, and MDA were significantly increased in a dose-dependent manner and/or a concentration-dependent manner in the groups treated with LPS （p0.05, compared with those of the control group）. Conclusion IL-15 may be involved in the pathological processes of peritoneal dialysis-related peritonitis. LPS can up-regulate the expression of IL-15, IL-6 and MDA in RPMCs, possibly relating to the peritonitis and oxidative stress.