摘要
目的探讨干扰paralemmin-3(PALM3)表达对脂多糖(LPS)诱导的肺泡上皮A549细胞炎性反应的影响。方法实验分为正常细胞组(未转染)、对照转染组(转染control siRNA)及PALM3siRNA转染组(转染针对PALM3的小干扰核糖核苷酸siRNA)三组;采用Western-blot法测PALM3蛋白表达水平,检验PALM3siRNA的干扰效率。三组细胞同时给予LPS刺激12h后,收集细胞培养上清液和总蛋白,ELISA法检测各组细胞培养上清中的TNF-α、IL-1β含量,同时用Western-blot法检测各组细胞中单免疫球蛋白白介素-1受体相关蛋白(SIGIRR)表达水平。结果特异性siRNA转染后成功干扰了PALM3在A549细胞中的表达;给予LPS刺激后,与正常细胞组及对照转染组相比,PALM3siRNA转染组细胞培养上清中TNF-α、IL-1β水平显著下降,同时PALM3siRNA转染组细胞中SIGIRR的表达上调。结论干扰PALM3的表达可减轻A549细胞对LPS诱导的炎性反应,其可能机制是通过上调Toll样受体/白介素-1受体(TIR)负调控分子SIGIRR的表达来实现的。
Objective To explore the effect of downregulation of paralemmin-3（PALM3）expression on the LPSinduced inflammatory reaction in alveolar epithelial cells（A549cells）.Methods The A549 cells were divided into three groups（PALM3siRNA group,control group and normal group）.In PALM3 siRNA group,the expression of PALM3 was down-regulated by PALM3 siRNA in A549cells;in control group,the cells were transfected with the control siRNA and any intervention was not administered in normal group.At 48 hafter transfection,the protein expression levels of PALM3 in A549cells were determined by Western-blot analysis.LPS was given to the cell culture medium after transfection.The culture supernatant and cellular total protein were collected at 12 hafter LPS-stimulation.The levels of TNF-αand IL-1βwere detected by ELISA.The protein expression levels of SIGIRR were also measured by Western-blot analysis.Results The expression of PALM3 was successfully suppressed by PALM3 siRNA.Compared with control and normal groups,downregulation of PALM3 expression could significantly decrease the release of TNF-αand IL-1βin A549 cells after LPS-stimulation.And,the protein levels of SIGIRR were significantly upregulated after LPS-stimulaition in PALM3 siRNA group.Conclusion Downregulation of PALM3 expression could attenuate the LPS-induced inflammation in alveolar epithelial cells.Upregulation of SIGIRR protein expression may be one of the possible mechanisms.
出处
《西部医学》
2015年第7期964-966,970,共4页
Medical Journal of West China
基金
国家自然科学基金(81300050)
海军总医院创新培养基金(CXPY201417)
