摘要
目的 研究在D-氨基半乳糖(D-Galn)/脂多糖(LPS)联合腹腔注射诱导小鼠急性肝衰竭模型中内质网应激介导肝细胞凋亡的作用,从而为急性肝衰竭的治疗提供思路。方法 以雄性巴比塞(BALB/c)小鼠为研究对象,腹腔注射D-GalN 800 mg/kg联合LPS 10μg/kg建立小鼠急性肝衰竭模型。动物实验分组:对照组(仅给予相应量的磷酸盐缓冲液)、急性肝衰竭模型组和4-丁酸苯酯(4-PBA)干预组(建模前6 h尾静脉注射)。同时将急性肝衰竭模型组按照给药时间进一步分为1 h、5 h、7 h三个亚组,蛋白质印迹法(Western blot)方法检测各亚组与对照组间内质网应激蛋白标志物78 kDa糖调节蛋白(GRP78)、内质网应激诱导的凋亡蛋白转录因子C/ERP的同源蛋白(CHOP),促凋亡因子Caspase-12和Caspase-3蛋白表达情况。检测血清中丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)水平评估肝脏功能,观察肝组织病理变化评价肝损伤程度。结果 Westernblot结果显示,与对照组比较,内质网应激蛋白标志物GRP78、内质网应激诱导的凋亡蛋白转录因子C/ERP的同源蛋白(CHOP)随着急性肝衰竭进展表达水平逐渐升高,促凋亡因子Caspase-12和Caspase-3蛋白在急性肝衰竭进展过程中表达亦升高。4-PBA抑制内质网应激可显著改善肝脏功能(血清ALT、AST水平明显下降,肝组织病理损伤明显改善);4-PBA干预组血清ALT[(733±103)U/L vs.(1 705±113)U/L]、AST[(1521±210)U/L vs.(3563±497)U/L]水平均显著低于急性肝衰竭模型组(P均<0.05),差异均有统计学意义(F=450.209,P=0.000;F=160.716,P=0.000);Westernblot研究结果显示,4-PBA对内质网应激进行干预,抑制肝细胞凋亡同时降低了凋亡蛋白Caspase-3表达。结论 内质网应激介导的肝细胞凋亡在D-氨基半乳糖/脂多糖联合注射诱发的小鼠急性肝衰竭中起关键作用,通过降低肝脏炎症反应和肝细胞凋亡,抑制内质网应对肝损伤起到改善作用。因此,干预
Objective To study the role of endoplasmic reticulum stress-induced hepatocyte apoptosis in acute liver failure model induced by D-galactosamine(D-Galn)and lipopolysaccharide(LPS)combined with intraperitoneal injection in mice,so as to provide new ideas for the treatment of acute liver failure.Methods Male BALB/c mice were injected intraperitoneally with 800 mg/kg D-Galn and 10μg/kg LPS to establish the model of acute liver failure.Animal experiment groups:the control group(corresponding volume phosphate buffered saline),the acute liver failure model group and Phenyl 4-butyrate(4-PBA)intervention group(tail vein injection 6 hours before modeling).At the same time,the acute liver failure model group was further divided into three subgroups according to the administration time:1,5 and 7 hours.Western Blotting method was used to detect the expression levels of ER stress protein markers 78kDa glucose regulatory protein(GRP78),ER stress-induced apoptosis protein transcription factor C/ERP homologous protein(CHOP)and hepatocyte apoptosis marker protein Caspase-12 and Caspase-3 in the liver tissue between subgroups and control group.Serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels were detected to evaluate liver function.Pathological changes of liver tissue were observed to evaluate the degree of liver injury.Results Comparing with control group,Westernblot results showed that the expression levels of ER stress protein markers GRP78 and ER stress-induced apoptosis protein transcription factor C/ERP homologous protein(CHOP)were increased with the progression of acute liver failure,and the expression levels of Caspase-12 and Caspase-3 also were increased during the progression of acute liver failure.The endoplasmic reticulum stress was inhibited by 4-PBA significantly improved liver function(serum ALT and AST levels were significantly decreased,and pathological injury of liver tissue was significantly improved).The levels of ALT[(733±103)U/L vs.(1705±113)U/L],AST[(1521±210)U/L vs.(3563±497
作者
冯之文
张文君
彭俊璐
戴大飞
鲍胜华
陈晓鹏
Feng Zhiwen;Zhang Wenjun;Peng Junlu;Dai Dafei;Bao Shenghua;Chen Xiaopeng(Department of Hepatobiliary surgery,Yijishan Hospital&the First Affiliated Hospital of Wannan Medical College,Wuhu,Anhui 241001,China)
出处
《齐齐哈尔医学院学报》
2023年第22期2106-2111,共6页
Journal of Qiqihar Medical University
基金
皖南医学院中青年重点科研基金项目(WK2022ZF21)
皖南医学院中青年科研基金项目(WK2018F10)。
关键词
急性肝衰竭
内质网应激
凋亡
炎症
Acute liver failure
Endoplasmic reticulum stress
Apoptosis
Inflammation
