丙酮酸激酶M2与miRNA-122在脓毒症相关AKI中的表达及其相关性研究  

Expression and correlation of pyruvate kinase M2 and miRNA-122 in sepsis associated AKI

作　　者：付周 李婷[2] 宋旭 李如欣 王凯丽 武宇涛 胡诺亚 权康丽 杜邦哲 FU Zhou;LI Ting;SONG Xu;LI Ru-xin;WANG Kai-li;WU Yu-tao;HU Nuo-ya;QUAN Kang-li;DU Bang-zhe(Undergraduates of Changzhi Medical College,Changzhi 046000,China;Department of Physiology,Changzhi Medical College,Changzhi 046000,China)

机构地区：[1]长治医学院本科生,山西长治046000 [2]长治医学院生理教研室,山西长治046000

出　　处：《海南医学院学报》2022年第24期1848-1854,共7页Journal of Hainan Medical University

基　　金：山西省高等学校大学生创新创业训练项目(20220846,2020385,2019399);国家自然科学基金项目(81902020);山西省应用基础研究计划项目(201801D221444);山西省高等学校科技创新项目(2019L0662);山西省卫生健康委科研课题(2018126);长治医学院学术技术带头人课题(XSQ201903)。

摘　　要：目的:探讨丙酮酸激酶M2(pyruvate kinase M2,PKM2)与miRNA-122在脓毒症相关急性肾损伤(sepsis-associated acute kidney injury,S-AKI)中的表达及其相关性。方法:通过盲肠结扎穿孔(cecal ligation and perforation,CLP)法诱导S-AKI小鼠模型,正常小鼠作为对照组。各模型组CLP术后4、8、12及24 h分别取血清及肾组织备用,生化分析仪检测各时间点尿素氮、肌酐水平。比色法检测各组小鼠血清及肾组织中的乳酸水平。实时荧光定量PCR(RT-qPCR)法检测各组小鼠肾组织中miRNA-122及pkm2 mRNA表达情况。Western blotting法检测各组小鼠肾组织中PKM2蛋白表达情况。采用Pearson法分析miRNA-122、PKM2与乳酸水平两两相关性。结果:(1)与对照组比较,CLP处理12、24 h后尿素氮、肌酐水平明显增加,差异具有统计学意义(P<0.05)。(2)与对照组比较,CLP处理12、24 h后血清及肾组织中的乳酸水平均明显升高,差异具有统计学意义(P<0.05)。(3)与对照组比较,CLP术后4 h miRNA-122在肾组织中表达开始降低,12~24 h明显降低,差异具有统计学意义(P<0.05)。与对照组比较,CLP术后4 h PKM2的mRNA与蛋白在肾组织中的表达开始增加,12~24 h明显增高,差异具有统计学意义(P<0.05)。(4)相关性分析显示miRNA-122与乳酸水平呈明显负相关(P<0.0001,r=-0.7167)。pkm2 mRNA与乳酸水平呈明显正相关(P<0.0001,r=0.6817)。miRNA-122与pkm2 mRNA的表达呈明显负相关(P<0.0001,r=-0.8122)。结论:在S-AKI中,miRNA-122的表达失调可能通过调控PKM2水平,促进有氧糖酵解与乳酸水平,加剧急性肾损伤的发生与发展。Objective:To investigate the expression and correlation of pyruvate kinase M2 and miRNA-122 in sepsis associated acute kidney injury(S-AKI).Methods:A mouse model of S-AKI was induced by cecal ligation and perforation(CLP),and normal mice were used in the control group.Serum and renal tissue were collected from each group,respectively,and the levels of blood urea nitrogen(BUN)and creatinine(Cr)were detected by biochemical analyzer.The levels of lactate in serum and kidney tissue of mice in each group were detected by colorimetric method.Real-time quantitative PCR(RT-qPCR)was used to detect the expression of miRNA-122 and pkm2 mRNA in kidney tissue in each group.Western blotting was used to detect the expression of PKM2 protein in kidney tissue in each group.Pearson’s method was used to analyze the pairwise correlation of miRNA-122,PKM2 and lactate levels.Results:(1)Compared with the control group,the levels of BUN and Cr increased significantly after 12 h and 24 h of CLP treatment(P<0.05).(2)Compared with the control group,after 12 h and 24 h of CLP treatment,the levels of lactate in the serum and kidney tissue of the mice were significantly higher than those in the control group(P<0.05).(3)Compared with the control group,the expression of miRNA-122 in renal tissue began to decrease at 4 h after CLP,and decreased significantly at 12-24 h(P<0.05).Compared with the control group,the expression of pKm2 mRNA and protein in renal tissue began to increase after 4 h of CLP,and increased significantly at 12-24 h(P<0.05).(4)Correlation analysis showed that miRNA-122 was significantly negatively correlated with lactate level(P<0.0001,r=-0.7167).There was a significant positive correlation between pkm2 mRNA and lactate level(P<0.0001,r=0.6817).There was a significant negative correlation between miRNA-122 and pkm2 mRNA expression(P<0.0001,r=-0.8122).Conclusion:In S-AKI,dysregulated expression of miRNA-122 may aggravate the occurrence and development of AKI by regulating the level of PKM2,and promoting aerobic glycolysis an

关 键 词：miRNA-122 PKM2 脓毒症 急性肾损伤 有氧糖酵解 

分 类 号：R459.7[医药卫生—急诊医学]