下调长链非编码RNA肺腺癌转移相关转录本-1调控星形胶质细胞水通道蛋白4表达改善脑缺血/再灌注损伤  

Regulating Aquaporin 4 Expression in Astrocytes and Improving Cerebral Ischemia Reperfusion Injury by Down-regulating LncRNA Metastasis-associated Lung Adenocarcinoma Transcript 1

作　　者：熊军 王宇[1] 郑丽芳 XIONG Jun;WANG Yu;ZHENG Li-fang(Department of Neurology,the Fourth Affiliated Hospital of Nanjing Medical University,Nanjing 210031,China;Department of Neurology,Yantian Hospital,Southern University of Science and Technology,Shenzhen 518000,China)

机构地区：[1]南京医科大学第四附属医院神经内科,南京210031 [2]南方科技大学盐田医院神经内科,深圳518000

出　　处：《中国临床神经科学》2023年第2期138-145,共8页Chinese Journal of Clinical Neurosciences

基　　金：广东省自然科学基金(编号:2020A151501287);深圳市科创委自然基金面上项目(编号:JCYJ20190808103401655)。

摘　　要：目的探讨下调长链非编码RNA(lncRNA)肺腺癌转移相关转录本-1(MALAT1)调控星形胶质细胞水通道蛋白4(AQP4)表达水平改善脑缺血/再灌注(CIR)损伤的机制。方法采用大脑中动脉闭塞法制作大鼠脑缺血模型:(1)将SD大鼠分成假手术组、缺血/再灌注组、阴性对照(Lv-NC)组和细胞感染慢病毒干扰载体(Lv-RNAi)组。进行大鼠神经功能评分,检测脑组织含水量,用qRT-PCR方法检测MALAT1表达,TTC染色法检测脑梗死体积,Western blot法检测AQP4表达,ELISA法检测TNF-α、IL-6、IL-1β水平。(2)分离大鼠星形胶质细胞分为:对照组、缺氧复氧(H/R)组、sh-NC组、sh-MALAT1组、sh-AQP4组、sh-MALAT1+Vector组和sh-MALAT1+AQP组。MTT法检测细胞增殖活性,检测TNF-α、IL-6、IL-1β、MALAT1和AQP4表达水平。结果(1)与假手术组比较,缺血/再灌注组MALAT1表达量升高,神经功能评分、脑组织含水量、脑梗死体积、AQP4蛋白表达量和TNF-α、IL-6、IL-1β含量升高(均P<0.05);与Lv-NC组比较,Lv-RNAi组MALAT1表达量降低,神经功能评分、脑组织含水量、脑梗死体积、AQP4蛋白表达量以及TNF-α、IL-6、IL-1β水平均降低(均P<0.05)。(2)星形胶质细胞中,与对照组比较,H/R组MALAT1和AQP4蛋白表达量升高,细胞增殖活性降低,TNF-α、IL-6、IL-1β水平增高(均P<0.05);与sh-NC组比较,sh-MALAT1组和sh-AQP4组AQP4蛋白表达量降低,细胞增殖活性升高,TNF-α、IL-6、IL-1β水平均降低(均P<0.05);与sh-MALAT1+Vector组比较,sh-MALAT1+AQP4组AQP4蛋白表达量升高,细胞增殖活性降低,TNF-α、IL-6、IL-1β水平均增高(均P<0.05)。结论下调MALAT1可能通过抑制星形胶质细胞AQP4表达进而改善CIR损伤。Aim To study the mechanism of regulating aquaporin 4(AQP4)expression in astrocytes and improving cerebral ischemia reperfusion(CIR)injury by down-regulating long-chain non-coding RNA(lncRNA)metastasis-associated lung adenocarcinoma transcript 1(MALAT1).Methods The middle cerebral artery occlusion method(thread embolism method)was used to make a rat model of cerebral ischemia.SD rats were divided into a sham operation group,a ischemia-reperfusion group(model),a negative control group(Lv-NC)and a cell-infected lentiviral interference vector group(LvRNAi).The expression of MALAT1 was detected by qRT-PCR,the neurological function of rats was scored,the water content of brain tissue was detected,and the volume of cerebral infarction was detected by TTC staining,Western blot was used to detect AQP4 protein expression,and ELISA was used to detect TNF-α,IL-6,and IL-1β.Rat astrocytes were isolated and divided into Control,H/R,sh NC,sh MALAT1,sh AQP4,sh MALAT1+Vector,sh MALAT1+AQP4 groups respectively.Cell proliferation activity was detected by MTT assay.MTT was used to detect the cells proliferation activities and the expression level of TNF-α,IL-6,IL-1β,MALAT,and AQP4 expression.Results Compared with the sham group,the expression of MALAT1 in the model group increased,and the neurological function score,brain tissue water content,cerebral infarction volume,AQP4 protein expression,and inflammatory factors TNF-α,IL-6,IL-1βincreased(P<0.05).Compared with the Lv-NC group,the expression of MALAT1 in the LvRNAi group was reduced,the neurological function score,brain tissue water content,cerebral infarction volume,AQP4 protein expression,and inflammatory factors TNF-α,IL-6,IL-1βdecreased(All P<0.05).Compared with the control group,the expression of MALAT1 and AQP4 protein in astrocytes of the H/R group increased,the cell proliferation activity decreased,and the inflammatory factors TNF-α,IL-6,and IL-1βsecreted by the cells increased(All P<0.05).Compared with the sh-NC group,the AQP4 protein expression in astrocytes in

关 键 词：肺腺癌转移相关转录本-1 长链非编码RNA 脑缺血/再灌注 星形胶质细胞 水通道蛋白4 

分 类 号：R743[医药卫生—神经病学与精神病学]