LncRNA NEAT1缺乏增加对乙酰氨基酚诱导的急性肝损伤的机制研究

Mechanism of LncRNA NEAT1 deficiency aggravating acetaminophen-induced acute liver injury

目的探讨长链非编码RNA(LncRNA)核富含丰富的转录本1(NEAT1)在对乙酰氨基酚(APAP)诱导小鼠急性肝损伤中的分子机制。方法8只C57BL/6小鼠随机分为实验组和对照组,每组各四只。通过腹腔注射APAP诱导急性肝损伤,进行谷丙转氨酶(ALT)、谷草转氨酶(AST)检测以及组织学分析来评估APAP诱导的肝损伤的程度。提取小鼠肝组织RNA检测LncRNA NEAT1表达变化。提取小鼠原代肝细胞并培养,用小干扰RNA敲低LncRNA NEAT1,并用APAP刺激小鼠原代肝细胞,通过检测细胞培养上清ALT和AST的表达量来评估肝细胞损伤程度。结果在体实验显示,与对照组相比,APAP处理组小鼠血清中的ALT[(42.75±2.3)U/L比(7565±763.3)U/L,(t=9.854,P<0.05)]和AST[(45.05±4.1)U/L比(7718±901.6)U/L,(t=8.510,P<0.05)]显著增加;APAP处理组小鼠肝组织LncRNA NEAT1的表达量比对照组表达增高3倍(P<0.05)。在体外实验中,原代肝细胞培养基加入APAP刺激24 h后提取RNA,LncRNA NEAT1的表达量显著增加;使用siRNA敲低肝细胞LncRNA NEAT1的表达并加入APAP刺激,与对照组相比,实验组肝细胞上清的ALT、AST显著增加。结论敲低LncRNA NEAT1的表达能够增加APAP诱导的急性肝损伤,提示LncRNA NEAT1可能具有防治APAP诱导的急性肝损伤的作用。

Objective The aim of this study was to investigate the molecular role of long non-coding ribonucleic acid(lncRNA)nuclear enriched abundant transcript 1(NEAT1)in the pathogenesis of acetaminophen(APAP)-induced acute liver injury.Methods In our study,8 C57BL/6 mice were randomly divided into experimental group and control group,with 4 mice in each group.Acute liver injury was induced by intraperitoneal injection of APAP.The mouse liver tissue RNA was extracted to detect the expression of lncRNA NEAT1.Mouse primary hepatocytes were extracted and cultured.In experimental group,hepatocytes were treated with small interfering ribonucleic acid(siRNA)to knock down lncRNA NEAT1,and then stimulated with APAP.The expression of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in cell culture supernatant is dectected to evaluate the degree of damage,together with histological analysis.Results In vivo,ALT(7565±763.3 vs 42.75±2.3,P<0.05)and AST(7718±901.6 vs 45.05±4.1,P<0.05)were significantly higher in the APAP-treated group than control group.The expression of lncRNA NEAT1 in liver tissues of the APAP-treated group was 3 times higher than that of control group(P<0.05).In vitro experiments,lncRNA NEAT1 expression was significantly increased after APAP stimulation in primary hepatocyte medium for 24 hours.ALT and AST in the supernatant of hepatocytes in the experimental group treated by siRNA and APAP were significantly higher than those in control group.Conclusion Our results indicate that knockdown of lncRNA NEAT1 can exacerbate APAP-induced acute liver injury,suggesting the potential benefit of lncRNA NEAT1 in the prevention and treatment of APAP-induced acute liver injury in the clinic.