自噬对胆总管结扎大鼠心肌损伤的影响  

Effects of autophagy on myocardial injury in rats with common bile duct ligation

作　　者：王晓玉 吕琳[1] 刘爱杰[1] 伦蕾 毕雯丽 董河[1] Wang Xiaoyu;Lyu Lin;Liu Aijie;Lun Lei;Bi Wenli;Dong He(Department of Anesthesiology,the Affiliated Hospital of Qingdao University,Qingdao 266000,Shandong,China)

机构地区：[1]青岛大学附属医院麻醉科,山东青岛266000

出　　处：《中华危重病急救医学》2025年第1期59-64,共6页Chinese Critical Care Medicine

基　　金：国家自然科学基金(81902001)。

摘　　要：目的探讨自噬对胆总管结扎(CBDL)大鼠心肌组织损伤的影响。方法将24只SPF级健康成年雄性SD大鼠按随机数字表法分为4组,每组6只。假手术(Sham)组大鼠仅游离胆总管但不结扎;CBDL组制备CBDL大鼠模型,模拟黄疸型心肌损伤过程;自噬抑制剂3-甲基腺嘌呤(3-MA)+CBDL组于制模前2 h腹腔注射3-MA 15 mg/kg,之后每隔1天注射1次;CBDL+自噬增强剂雷帕霉素(Rapa)组于制模后0.5 h腹腔注射Rapa 1 mg/kg,之后每隔1天注射1次。各组均于术后2周处死大鼠,取下腔静脉血,使用全自动动物生化分析仪检测血清总胆红素(TBil)、丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、乳酸脱氢酶(LDH)和肌酸激酶同工酶(CK-MB);采用比色法检测血清氧化应激标志物超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量。取大鼠心肌组织,苏木素-伊红(HE)染色后,光镜下观察病理学改变;醋酸铀-枸橼酸铅双染后,透射电镜下观察自噬体;采用蛋白质免疫印迹试验(Western blotting)检测心肌组织中自噬相关蛋白表达。结果与Sham组相比,CBDL组大鼠血清SOD活性显著下降,血清MDA、TBil、ALT、AST、LDH、CK-MB水平显著升高;心肌组织中自噬相关蛋白Beclin-1和微管相关蛋白1轻链3-Ⅱ/Ⅰ(LC3-Ⅱ/Ⅰ)蛋白表达显著上调,p62蛋白表达显著下调;透射电镜下可见自噬体,病理形态学观察可见心肌组织中灶状坏死及炎症细胞浸润,心肌间质小血管扩张,并出现心肌纤维变性。与CBDL组比较,给予自噬抑制剂3-MA预处理可以减轻大鼠心肌组织损伤,减少炎症细胞浸润,减轻心肌间质血管扩张,减小心肌纤维化面积,且透射电镜下可见自噬体明显减少;血清TBil、ALT、AST进一步升高〔TBil(μmol/L):184.40±6.74比120.70±16.93,ALT(U/L):501.10±62.18比178.80±22.30,AST(U/L):806.50±76.92比275.50±55.81,均P<0.01〕,SOD、MDA、LDH、CK-MB活性显著下降〔SOD(kU/L):85.00±5.29比107.50±7.86,MDA(μmol/L):10.72±0.93比15.06±1.88,LDH(U/LObjective To investigate the impact of autophagy on cardiac tissue injury following common bile duct ligation(CBDL)in rats.MethodsTwenty-four SPF grade healthy adult male Sprague-Dawley(SD)rats were randomly divided into four groups,with 6 rats in each group.The sham-operated(Sham)group underwent only dissection of the common bile duct without ligation.The CBDL group underwent CBDL to simulate jaundice-induced myocardial injury.The autophagy inhibitor 3-methyladenine(3-MA)+CBDL group was intraperitoneally injected with 15 mg/kg 3-MA 2 hours before modeling,and then injected once every other day.The CBDL+autophagy enhancer rapamycin(Rapa)group was intraperitoneally injected with Rapa 1 mg/kg 0.5 hour after modeling,and then injected once every other day.The rats in each group were sacrificed 2 weeks after surgery,and blood was taken from the inferior vena cava.Serum total bilirubin(TBil),alanine transaminase(ALT),aspartate transaminase(AST),lactate dehydrogenase(LDH),and MB isoenzyme of creatine kinase(CK-MB)were detected by using a fully automated animal biochemical analyzer.Serum oxidative stress marker superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were detected by colorimetric assay.The heart tissues of rats were taken and pathological changes were observed under a light microscope after hematoxylin-eosin(HE)staining.Transmission electron microscope was used to observe autophagosomes after double staining with uranyl acetate and lead citrate.The expressions of autophagy-related proteins were detected using Western blotting.ResultsCompared with the Sham group,the serum SOD activity of rats in the CBDL group was significantly decreased,while the serum MDA,TBil,ALT,AST,LDH,and CK-MB were significantly increased;the expressions of autophagy-related proteins Beclin-1 and microtubule-associated protein 1 light chain 3-Ⅱ/Ⅰ(LC3-Ⅱ/Ⅰ)were significantly increased,and p62 protein expression was significantly decreased.Autophagosomes were seen under electron microscopy in the CBDL group,and cardi

关 键 词：胆总管结扎 心肌损伤 自噬 3-甲基腺嘌呤 雷帕霉素 

分 类 号：R614[医药卫生—麻醉学]