抑制自噬加重脑死亡状态下肝细胞损伤  

Inhibiting Autophagy Increases Hepatocellular Injury Under Brain Death State

作　　者：李月霞[1] 石慧娟[1] 曹胜利[2] LI Yuexia;SHI Huijuan;CAO Shengli(Department of Critical Care Medicine,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 453000,China;Department of Hepatobiliary and Pancreatic Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 453000,China)

机构地区：[1]郑州大学第一附属医院重症医学科,河南郑州453000 [2]郑州大学第一附属医院肝胆胰外科,河南郑州453000

出　　处：《河南医学研究》2024年第15期2698-2701,共4页Henan Medical Research

基　　金：NSFC-河南联合基金(U2004122)。

摘　　要：目的探讨凋亡和自噬在脑死亡状态下肝细胞损伤中的作用。方法将40只SD大鼠随机分为4组:对脑死亡组采用缓慢间歇颅内加压法诱导脑死亡,并且维持脑死亡状态6 h;对假手术组进行颅内置管,但是不进行脑死亡的诱导;对脑死亡+自噬抑制剂组诱导脑死亡前1 h腹腔注射自噬抑制剂3-甲基腺嘌呤(3-MA),建立并维持脑死亡状态6 h;对脑死亡+空白溶剂组诱导脑死亡前1 h腹腔注射空白溶剂二甲基亚砜(DMSO),建立并维持脑死亡状态6 h。造模完成后,取大鼠静脉血及肝脏标本。通过荧光定量qPCR检测Caspase-3 mRNA表达水平;通过Western blot检测LC3及Caspase-3蛋白表达水平;通过免疫组化检测Caspase-3蛋白在肝组织中分布和表达。结果与假手术组相比,脑死亡组中凋亡相关基因Caspase-3表达上升,肝细胞凋亡加重;自噬相关基因LC3表达上调(P<0.05);与脑死亡+空白溶剂组相比,脑死亡组凋亡相关基因及自噬相关基因表达差异无统计学意义(P>0.05);与脑死亡+空白溶剂组相比,脑死亡+自噬抑制剂组中LC3表达下调(P<0.05),凋亡相关基因Caspase-3表达增加(P<0.05),并且肝细胞凋亡增加(P<0.05)。结论细胞凋亡参与并介导脑死亡状态下的肝细胞损伤,自噬能减轻脑死亡状态下的肝细胞损伤。Objective To investigate the role of apoptosis and autophagy on hepatic injuryunder brain death state.Methods The 40 SD rats were randomly divided into 4 groups:the brain death group was induced by slow intermittent intracranial compression and maintained in a state of brain death for 6 hours,perform intracranial catheterization on the sham surgery group,but did not induce brain death,inject autophagy inhibitor 3-methyladenine(3-MA)intraperitoneally into the brain death+autophagy inhibitor group 1 hour before inducing brain death,and establish and maintain a state of brain death for 6 hours,for the brain death+blank solvent group intraperitoneal injection of blank solvent dimethyl sulfoxide(DMSO)was administered 1 hour before inducing brain death,and the brain death state was established and maintained for 6 hours.After the molding was established,the venous blood and liver specimens were taken from rats.qPCR was used to detect the expression of Caspase-3 mRNA.Western blot was used to detect the expression of LC3 and Caspase-3 protein.Immunohistochemistry was used to detect the distribution and expression of Caspase-3 in liver tissues.Results Compared with sham surgery group,the expression of apoptosis-related gene Caspase-3 was up-regulated and the apoptosis of hepatocytes increased in brain death group,the expressions of autophagy-related genes LC3 were up-regulated(P<0.05).Compared with the DMSO group,there was no statistical difference in apoptosis-related genes and autophagy-related genes in brain death group(P>0.05).Compared with the DMSO group,the expression of LC3 in 3-MA group was down-regulated(P<0.05),the expression of apoptosis-related gene Caspase-3 was up-regulated(P<0.05),and the apoptosis of hepatocytes increased(P<0.05).Conclusion Apoptosis participates in and mediates the hepatocellular injury under brain death,and autophagy attenuates hepatocellular injury underbrain death state.

关 键 词：脑死亡 肝细胞凋亡 自噬 

分 类 号：R31[医药卫生—基础医学]