下调长链非编码RNA MALAT1表达对小胶质细胞活化的影响  

Effect of knockdown of long noncoding RNA metastasis associated lung adenocarcinoma transcript 1 on microglial activation

作　　者：张军玲 潘海英[1] 徐敏[1] 李晓云[1] 牛建一[1] Zhang Junling;Pan Haiying;Xu Min;Li Xiaoyun;Niu Jianyi(Department of Neurology, Yidu Central Hospital of Weifang City, Weifang 262500, Chin)

机构地区：[1]潍坊市益都中心医院神经内科,262500

出　　处：《中华神经医学杂志》2018年第5期445-449,共5页Chinese Journal of Neuromedicine

摘　　要：目的 探讨下调长链非编码RNA人肺腺癌转移相关转录本1（MALAT1）表达对小胶质细胞活化的影响及其可能的机制. 方法 体外培养BV2小胶质细胞,并将其分为正常对照组（不作任何处理）、脂多糖（LPS）处理组（用含100 ng/mL LPS的培养液培养）、MALAT1干扰组（转染MALAT1干扰序列＋用含100 ng/mL LPS的培养液培养）和对照序列组（转染对照序列＋用含100ng/mL LPS的培养液培养）,采用逆转录-实时定量PCR（RT-qPCR）检测细胞中MALAT1 mRNA的表达,Western blotting检测细胞中核因子-κB（NF-κB）及其抑制蛋白（IκB-α）蛋白的表达,ELISA法检测细胞上清液中IL-1β、IL-6、IL-10和肿瘤坏死因子-α（TNF-α）水平,Griess法检测细胞上清液中一氧化氮（NO）释放量. 结果 与正常对照组比较,LPS处理组、对照序列组和MALAT1干扰组细胞中MA LA T1 mRNA相对表达量均明显升高,NF-κB蛋白相对表达量均明显升高,IκB-α蛋白相对表达量均明显降低,细胞上清液中IL-1β、IL-6、IL-10、TNF-α水平及NO释放量均明显升高,差异均有统计学意义（P＜0.05）;与LPS处理组、对照序列组比较,MALAT1干扰组细胞中MALA T1 mRNA相对表达量明显降低,NF-κB蛋白相对表达量明显降低,IκB-α蛋白相对表达量明显升高,细胞上清液中IL-1β、IL-6、IL-10、TNF-α水平及NO释放量均明显降低,差异均有统计学意义（P＜0.05）. 结论 下调MALAT1表达可抑制LPS诱导的BV2小胶质细胞活化导致的炎症反应,其机制可能与抑制NF-κB信号通路有关.Objective To investigate the effect of down-regulation of human lung adenocarcinoma metastasis-associated transcript 1 （MALAT1） on microglial activation and its possible mechanism.Methods BV2 microglial cells were cultured in vitro and divided into a normal control group （no treatment）,a lipopolysaccharide （LPS） treatment group （cultured with 100 ng/mL LPS）,a MALAT1 interference group （transfected with MALAT1 interference sequence ＋ cultured with 100 ng/mL LPS） and a control sequence group （transfected with control sequence ＋ cultured with 100 ng/mL LPS）.Real-time fluorescence quantitative PCR was used to detect the expression ofMA LA T1 mRNA in the cells.Western blotting was used to detect the expression of NF-κB protein and IκB-α protein in the cells.Enzyme linked immunosorbent assay （ELISA） was used to detect the levels of IL-1β,IL-6,IL-10 and TNF-α in each group.Nitric acid reduction method （Griess method） was used to detect nitric oxide （NO） concentration in each group.Results The relative expression levels of MALA T1 mRNA and NF-κB protein were significantly increased,the relative expression level of IκB-α protein was significantly decreased,the levels ofIL-1β,IL-6,IL-10 and TNF-α and the release of NO in the cell supernatant were significantly increased in the LPS treatment,MALAT1 interference and control sequence groups than in the normal control group （P＜0.05）.The relative expression levels of MALAT1 mRNA and NF-κB protein were significantly decreased,the expression level of IκB-α protein was significantly increased,the levels ofIL-1β,IL-6,IL-10 and TNF-α and the release of NO in the cell supematant were significantly decreased in the MALAT1 interference group than in the LPS treatment and control sequence groups （P＜0.05）.Conclusions Down-regulation of MALA T1 gene expression may inhibit the inflammatory response induced by LPS-induced BV2 cell activation.The mechanism might be related to the inhibition of NF-κB signaling pathway.

关 键 词：人肺腺癌转移相关转录本1 小胶质细胞 炎症反应 核因子-ΚB信号通路 长链非编码RNA 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]