白花蛇舌草提取物通过AMPK/ATG5信号通路对急性胰腺炎大鼠肺损伤的保护作用机制研究  

Study on protective mechanism of hedyotis diffusa extract on lung injury in ratswith acute pancreatitis through AMPK/ATG5 signaling pathway

作　　者：哈宗兰 马丽娜 马琼 甘桂芬[1] HA Zonglan;MA Lina;MA Qiong;GAN Guifen(Department of Critical Medicine,Affiliated Hospital of Qinghai University,Xining 810000,China;Department of Radiology,Affiliated Hospital of Qinghai University,Xining 810000,China)

机构地区：[1]青海大学附属医院重症医学科,西宁810000 [2]青海大学附属医院放射科,西宁810000

出　　处：《中国免疫学杂志》2024年第2期348-354,共7页Chinese Journal of Immunology

摘　　要：目的:通过5’-单磷酸腺苷活化蛋白激酶(AMPK)/自噬相关蛋白5(ATG5)信号通路,探讨白花蛇舌草提取物减轻急性胰腺炎(AP)大鼠肺损伤的机制。方法:建立AP肺损伤大鼠模型,随机分为模型组、提取物(白花蛇舌草提取物)组、3-MA(自噬抑制剂3-甲基腺嘌呤)组、AICAR(AMPK激活剂)组、提取物+AICAR组,每组15只,另取15只大鼠作为假手术组;ELISA检测血清淀粉酶(AMY)、IL-1β、IL-6、IL-18水平;检测大鼠腹水量及肺湿/干重比(W/D);HE观察胰腺及肺组织病理损伤;Western blot检测溶酶体相关膜蛋白2(LAMP2)、自噬底物p62、IL-1β前体蛋白(pro-IL-1β)、胰蛋白酶原活化肽(TAP)、AMPK、p-AMPK、ATG5、自噬标志物LC3-Ⅱ/Ⅰ、泛素特异性蛋白酶10(UPS10)表达。结果:与假手术组相比,模型组大鼠腹水量、肺W/D、胰腺和肺组织病理损伤评分、血清AMY、IL-1β、IL-6、IL-18水平、胰腺组织p62、TAP、pro-IL-1β表达、肺组织pAMPK/AMPK、ATG5、LC3-Ⅱ/Ⅰ、UPS10、pro-IL-1β表达均升高(P<0.05),胰腺和肺组织LAMP2蛋白表达降低(P<0.05);模型大鼠经白花蛇舌草提取物或自噬抑制剂3-MA干预后,上述指标均得到显著改善(P<0.05),且白花蛇舌草提取物的改善效果优于3-MA(P<0.05);而AMPK激活剂AICAR可削弱白花蛇舌草提取物对AP大鼠肺损伤的改善作用(P<0.05)。结论:白花蛇舌草提取物可通过抑制AMPK/ATG5信号通路减轻炎症反应、降低自噬水平改善AP大鼠肺损伤。Objective:To explore mechanism of hedyotis diffusa extract attenuating acute pancreatitis(AP)lung injury in rats from 5'-adenosine monophosphate activated protein kinase(AMPK)/autophagy-related protein 5(ATG5)signaling pathway.Methods:AP lung injury rat model was established,and rats were randomly divided into model group,extract(hedyotis diffusa extract)group,3-MA(autophagy inhibitor 3-methyladenine)group,AICAR(AMPK activator)group and extract+AICAR group,with 15 rats in each group,and another 15 rats were selected as sham operation group.ELISA was performed to determine levels of serum amylase(AMY),IL-1β,IL-6 and IL-18;amount of ascites and proportion of lung wet/dry(W/D)in rats were determined;HE was performed to observe pathological damage of pancreas and lung tissues;Western blot was performed to determine lysosomal-associated membrane protein 2(LAMP2),autophagy substrate p62,IL-1βprecursor protein(pro-IL-1β),trypsinogen activation peptide(TAP),AMPK,p-AMPK,ATG5,autophagy marker LC3-Ⅱ/Ⅰ,ubiquitin-specific protease 10(UPS10)expressions.Results:Compared with sham operation group,ascites volume,lung W/D,pancreas and lung tissue pathological damage scores,serum AMY,IL-1β,IL-6,IL-18 levels,pancreatic tissue p62,TAP,pro-IL-1βexpressions,lung tissue p-AMPK/AMPK,ATG5,LC3-Ⅱ/Ⅰ,UPS10,pro-IL-1βexpressions of rats in model group were increased(P<0.05),while LAMP2 protein expression in pancreas and lung tissue was decreased(P<0.05);after model rats were intervened by hedyotis diffusa extract or autophagy inhibitor 3-MA,the above indexes were significantly improved(P<0.05),and improvement effect of hedyotis diffusa extract was better than 3-MA(P<0.05);while AMPK activator AICAR could attenuate improvement effect of hedyotis diffusa extract on lung injury in AP rats(P<0.05).Conclusion:Hedyotis diffusa extract can reduce inflammatory response and autophagy level by inhibiting AMPK/ATG5 signaling pathway,and improve lung injury in AP rats.

关 键 词：白花蛇舌草提取物 胰腺炎 肺损伤 5’-单磷酸腺苷活化蛋白激酶 自噬相关蛋白5 炎症-自噬 

分 类 号：R576[医药卫生—消化系统]