中药材2024，Vol.47Issue(10)：2584-2590,7.DOI:10.13863/j.issn1001-4454.2024.10.030
清解化攻方调节PINK1/Parkin介导的线粒体自噬对重症急性胰腺炎大鼠肠道炎症反应的影响
Effect of Qingjie Huagong Prescription on Intestinal Inflammation in Rats with Severe Acute Pancreatitis by Regulating PINK1/Parkin-Mediated Mitochondrial Autophagy
摘要
Abstract
Objective:To investigate the mechanism of Qingjie huagong prescription regulating intestinal injury in severe acute pan-creatitis by regulating PINK1/Parkin-mediated mitochondrial autophagy.Methods:SD male rats were randomly divided into normal con-trol group,model group,Qingjie huagong prescription group,Qingjie huagong prescription with CCCP(mitochondrial autophagy activa-tor)group,Mdivi-1(mitochondrial autophagy inhibitor)group.The intestinal injury model of severe acute pancreatitis was established with 5%sodium taurocholate retrograde pancreatitis duct.After successful modeling,rats in each drug administration group were given corresponding drugs,and the rats in normal control group and model group were given corresponding volume of normal saline.The con-tents of serum lipase,amylase,TNF-α and IL-6 in the homogenate of ileal tissue of rats were detected by ELISA after 24 hours adminis-tration.The pathological changes of pancreas and ileum were observed by HE staining.The content of ATP in the homogenate of ileum was determined by colorimetry.The ultrastructure of mitochondria in rat ileum was observed by transmission electron microscopy.The immunocolocalization of LC3B and TOMM20 in ileum was observed by immunofluorescence staining.The mRNA and protein expressions of PINK1,Parkin,LC3,TNF-α and IL-6 in rat ileum were detected by Real-time PCR and Western Blot.Results:Compared with the normal control group,the structure of pancreas and ileum in model group was disordered,accompanied by a large number of inflammato-ry cell infiltration,and the pathological injury was serious.The contents of serum amylase,lipase and TNF-α and IL-6 in ileal homoge-nate were significantly increased,while the content of ATP in ileal homogenate was significantly decreased(P<0.05).Most of the mito-chondria were blurred,dilated,vacuolated and dissolved,the number of mitochondria decreased,and a large number of autophagosomes were observed by fluoroscopic electron microscopy.The double labeling of immunofluorescence LC3B and TOMM20 was significantly in-creased.The mRNA and protein expressions of PINK1,Parkin,LC3Ⅱ/Ⅰ,TNF-α and IL-6 were significantly increased(P<0.05).Compared with the model group,the pathological injury of pancreas and ileal tissue in Qingjie huagong prescription group and Mdivi-1 group was improved.The levels of serum amylase,lipase,TNF-α and IL-6 in the homogenate of ileum were significantly decreased,while the level of ATP in ileum was significantly increased(P<0.05).The number of normal mitochondria in ileum was increased by X-ray electron microscopy.Most mitochondria were basically intact in morphology and structure.A few mitochondria were still swollen and the mitochondrial fusion division was less,a few autophagosomes were seen.The double labeling of immunofluorescence LC3B and TOMM20 was significantly decreased,and the mRNA and protein expressions of PINK1,Parkin,LC3,TNF-α and IL-6 in ileum were significantly decreased(P<0.05).There was no significant difference between the indexes in Qingjie huagong prescription group and Mdivi-1 group(P>0.05).There was no significant difference between the indexes in Qingjie huagong prescription with CCCP group and model group(P>0.05).Conclusion:Qingjie huagong prescription may alleviate intestinal inflammatory injury in severe acute pancreatitis by inhibi-ting excessive activation of mitochondrial autophagy.关键词
重症急性胰腺炎肠损伤/清解化攻方/线粒体自噬/PINK1/Parkin通路/肠道炎症反应Key words
Severe acute pancreatitis with intestinal injury/Qingjie huagong prescription/Mitochondrial autophagy/PINK1/Par-kin Pathway/Intestinal inflammatory response分类
医药卫生引用本文复制引用
李慧,李德文,姚春,安晓霞,张冰玉,唐曦平,陈国忠..清解化攻方调节PINK1/Parkin介导的线粒体自噬对重症急性胰腺炎大鼠肠道炎症反应的影响[J].中药材,2024,47(10):2584-2590,7.基金项目
高水平中医药重点学科建设项目-中医内科学(zyyzdxk-2024016-01-01) （zyyzdxk-2024016-01-01）
国家自然科学基金项目(8216150526) （8216150526）
广西中医药大学研究生教育创新计划项目(YCBXJ2023013) （YCBXJ2023013）
广西中医药大学"桂派中医药传承创新团队"A类资助项目(2022A001) （2022A001）
广西自然科学基金面上项目(2020GXNSFAA297062) （2020GXNSFAA297062）
国家中医优势专科建设(2024010) （2024010）
