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目的 :观察白细胞介素 10 (IL 10 )对白细胞介素 1β(IL 1β)诱导的人系膜细胞 (HMC)胞质型磷脂酶A2 (cPLA2 )基因和蛋白表达及其前列腺素E2 (PGE2 )释放的影响。方法 :利用体外培养的HMC ,设立对照组、IL 10处理组、IL 1β处理组及IL 10+IL 1β处理组 ;采用放免法检测培养上清中PGE2 ,应用逆转录 多聚酶链反应 (RT PCR)和蛋白质印迹 (WesternBlot)检测cPLA2mRNA和蛋白水平。结果 :正常情况下 ,HMC低水平表达cPLA2 mRNA和蛋白并释放少量的PGE2 ;IL 1β能显著上调HMCPGE2释放及cPLA2 mRNA和蛋白的表达 (P值均 <0 .0 1) ;IL 10对基础状态下的HMCPGE2 释放及cPLA2 mRNA和蛋白表达无明显影响 (P值均 >0 .0 5 ) ,但可呈剂量依赖性地下调IL 1β诱导的PGE2 释放及cPLA2 mRNA和蛋白表达 (P值均 <0 .0 1)。结论 :IL 10抑制IL 1β诱导的HMCPGE2 释放及cPLA2 表达 ,提示IL 10对HMC具有多方面抗炎作用。
Objective: To investigate the effect of interleukin 10 (IL 10) on the gene and protein expression of cytoplasmic phospholipase A2 (cPLA2) and its prostaglandin E2 (PGE2) - induced human interleukin - 1β The impact of release. Methods: The control group, IL 10 treatment group, IL 1β treatment group and IL 10 + IL 1β treatment group were established by using HMC cultured in vitro. PGE2 in the culture supernatant was detected by radioimmunoassay. Reverse transcription-polymerase chain reaction (RT PCR) And Western blotting to detect cPLA2 mRNA and protein levels. Results: Under normal conditions, HMC expressed low levels of cPLA2 mRNA and protein and released a small amount of PGE2; IL 1β significantly up-regulated the release of HMCPGE2 and the expression of cPLA2 mRNA and protein (all P <0.01) (P <0.05). However, the release of HMIPGE2 and the expression of cPLA2 mRNA and protein were not significantly affected (P <0.05), but downregulated IL-1β-induced PGE2 release and cPLA2 mRNA and protein expression .0 1). CONCLUSION: IL-10 inhibits IL-1β-induced HMCPGE2 release and cPLA2 expression, suggesting that IL 10 has multiple anti-inflammatory effects on HMC.