lncRNA MIAT通过调节miR-338-3p/THBS1轴促进脓毒症引起的急性肾损伤  

lncRNA MIAT promotes sepsis-induced acute kidney injury by regulating the miR-338-3p/THBS1 axis

作　　者：刘霄岩 赵秋兰 代江娜 LIU Xiaoyan;ZHAO Qiulan;DAI Jiangna(Department of Intensive Care Medicine,the NO.2 Hospital of Baoding,Baoding,Hebei 071000,China)

机构地区：[1]保定市第二医院重症医学科,河北保定071000

出　　处：《国际检验医学杂志》2025年第6期681-688,共8页International Journal of Laboratory Medicine

基　　金：河北省中医药类科学研究课题计划项目(2023436)。

摘　　要：目的探讨长链非编码RNA(lncRNA)心肌梗死相关转录物(MIAT)是否通过调节微小RNA-338-3p(miR-338-3p)/血小板凝血酶蛋白-1(THBS1)轴促进脓毒症引起的急性肾损伤(AKI)。方法用盲肠结扎穿孔法建立脓毒症AKI模型,将大鼠分为对照组和脓毒症AKI组,每组10只。采用实时荧光定量PCR(qPCR)检测肾组织lncRNA MIAT、miR-338-3p和THBS1基因表达水平,全自动生化分析仪检测血清尿素氮(BUN)和肌酐(Cre)水平。采用脂多糖(LPS)诱导大鼠肾小管上皮NRK-52E细胞建立脓毒症AKI细胞模型,将NRK-52E细胞分为CK组、LPS组、LPS+si-NC组、LPS+si-lncRNA MIAT组、LPS+si-lncRNA MIAT+inhibitor NC组、LPS+si-lncRNA MIAT+miR-338-3p inhibitor组、LPS+si-lncRNA MIAT+oe-NC组、LPS+si-lncRNA MIAT+oe-THBS1组。采用qPCR检测各组细胞lncRNA MIAT、miR-338-3p和THBS1基因表达水平,细胞计数试剂盒8检测各组细胞增殖情况,流式细胞术检测各组细胞凋亡率,酶联免疫吸附试验检测各组乳酸脱氢酶(LDH)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-10、丙二醛(MDA)水平及超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性,蛋白质印记法检测细胞中NOD样受体热蛋白结构域相关蛋白3(NLRP3)、半胱氨酸天冬氨酸蛋白酶1(Caspase-1)、裂解凋亡蛋白酶-3(cleaved caspase-3)、THBS1蛋白表达。验证miR-338-3p与lncRNA MIAT和THBS1的靶向关系。结果脓毒症AKI组大鼠BUN、Cre、lncRNA MIAT、THBS1基因表达水平升高(P<0.05),miR-338-3p表达水平降低(P<0.05)。与CK组比较,LPS组和LPS+si-NC组脓毒症AKI细胞中lncRNA MIAT表达、THBS1基因表达、凋亡率和IL-6、LDH、TNF-α、MDA水平升高(P<0.05),NLRP3、Caspase-1、cleaved caspase-3、THBS1蛋白表达水平升高(P<0.05),miR-338-3p表达、A_(450)(24、48 h)值、IL-10水平降低(P<0.05),CAT和SOD活性降低(P<0.05)。与LPS+si-NC组比较,LPS+si-lncRNA MIAT组脓毒症AKI细胞中lncRNA MIAT表达、THBS1基因表达、凋亡率和IL-6、LDH、TNF-α、MDA水平降低Objective To investigate whether long non-coding RNA(lncRNA)myocardial infarct-associated transcription factor(MIAT)promotes sepsis-induced acute kidney injury(AKI)by regulating microRNA-338-3p(miR-338-3p)/platelet thromboplastin-1(THBS1)axis.Methods Septic AKI model was established by cecal ligation and puncture.The rats were divided into control group and sepsis AKI group,with 10 rats in each group.The expression levels of lncRNA MIAT,miR-338-3p and THBS1gene in renal tissue were detected by quantitative real-time PCR(qPCR).The levels of serum urea nitrogen(BUN)and creatinine(Cre)were detected by automatic biochemical analyzer.Renal tubular epithelial NRK-52E cells were induced by lipopolysaccharide(LPS)to establish a cell model of sepsis induced AKI.NRK-52E cells were divided into CK group,LPS group,LPS+si-NC group,LPS+si-lncRNA MIAT group,LPS+si-lncRNA MIAT+inhibitor NC group,LPS+si-lncRNA MIAT+miR-338-3p inhibitor group,LPS+si-lncRNA MIAT+oe-NC group,LPS+si-lncRNA MIAT+oe-THBS1 group.qPCR was used to detect the expression levels of lncRNA MIAT,miR-338-3p and THBS1 gene in each group.Cell counting kit 8 was used to detect cell proliferation in each group.Flow cytometry was used to detect the apoptosis rate of each group.Enzyme-linked immunosorbent assay was used to detect the levels of lactate dehydrogenase(LDH),tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-10,malondialdehyde(MDA)and the activities of superoxide dismutase(SOD)and catalase(CAT)in each group.The expression of NOD-like receptor pyrin domain-containing protein 3(NLRP3),Caspase-1,cleaved caspase-3 and THBS1 protein were detected by Western blot.The targeting relationship between miR-338-3p and lncRNA MIAT and THBS1 was verified.Results The expression levels of BUN,Cre,lncRNA MIAT and THBS1 gene were increased(P<0.05),and the expression level of miR-338-3p was decreased in sepsis AKI group(P<0.05).Compared with CK group,the expression of lncRNA MIAT,THBS1 gene,apoptosis rate and the levels of IL-6,LDH,TNF-αand MDA were significantly increa

关 键 词：长链非编码RNA心肌梗死相关转录物 微小RNA-338-3p/血小板凝血酶蛋白-1轴 脓毒症 急性肾损伤 

分 类 号：R692[医药卫生—泌尿科学] R459.7[医药卫生—外科学]