摘要
目的探讨清热利胆的经典中药方剂——茵陈蒿汤(YCHT)对牛磺胆酸钠诱导的大鼠重症急性胰腺炎(SAP)的保护作用及其机制。方法30只SD大鼠随机分成假手术(SO)组、SAP模型组和YCHT(4.0 g/kg)治疗组,每组10只。造模成功24 h后,留取大鼠胰腺组织和血浆待检测。HE染色观察胰腺病理损伤情况;ELISA法检测血浆淀粉酶、TNFα和IL-1β水平;免疫荧光染色检测LC-3蛋白的荧光强度,TUNEL检测细胞凋亡情况。Western Blot检测胰腺组织LC-3、Beclin-1、X连锁凋亡抑制蛋白(XIAP)、Caspase-3和NF-κB蛋白表达。实时荧光定量PCR检测lncRNA PVT1和miRNA-30a-5p的表达水平。采用单因素方差分析和Tukey’s检验用于分析多个独立样本之间的差异。结果YCHT能明显减轻SAP大鼠胰腺组织水肿、坏死、出血及炎性细胞浸润等病理学损伤。与SO组相比,SAP模型组大鼠血浆淀粉酶、炎性因子TNFα和IL-1β水平显著升高,YCHT治疗后血浆淀粉酶及TNFα和IL-1β水平显著降低(P值均<0.05)。与SO组相比,SAP模型组LC-3Ⅱ/LC-3Ⅰ比值及Beclin-1、XIAP、Caspase-3和NF-κB蛋白表达明显上调,YCHT治疗组LC-3Ⅱ/LC-3Ⅰ、Beclin-1、XIAP和NF-κB的表达水平较SAP模型组显著降低,Caspase-3水平显著升高(P值均<0.05)。与SO组相比,SAP模型组大鼠胰腺lncRNA PVT1表达显著升高,miRNA-30a-5p表达显著降低(P值均<0.05);与SAP模型组相比,YCHT显著降低了lncRNA PVT1的表达,增加了miRNA-30a-5p的表达(P值均<0.05)。结论lncRNA PVT1/miRNA-30a-5p介导的细胞自噬和凋亡可能是YCHT治疗SAP的一个药物靶点,这为进一步开发中药方剂YCHT治疗SAP提供了实验基础和理论依据。
Objective To investigate the protective effect and mechanism of Artemisia capillaris Thunb.decoction(YCHT),a classic heat-clearing and cholagogic traditional Chinese medicine(TCM)prescription,on rats with severe acute pancreatitis(SAP)induced by sodium taurocholate.Methods A total of 30 Sprague-Dawley rats were randomly divided into sham-operation(SO)group,SAP model group,and YCHT(4.0 g/kg)treatment group,with 10 rats in each group.At 24 hours after successful modeling,pancreatic tissue and plasma samples were collected for analysis.HE staining was used to observe pathological injury of the pancreas;ELISA was used to measure the plasma levels of amylase,tumor necrosis factor-α(TNFα),and interleukin-1β(IL-1β);immunofluorescent staining was used to measure the fluorescence intensity of LC-3 protein,and TUNEL was used to measure cell apoptosis.Western blot was used to measure the protein expression of LC-3,Beclin-1,X-linked inhibitor of apoptosis protein(XIAP),caspase-3,and nuclear factor-kappa B(NF-κB)in the pancreas,and quantitative real-time PCR was used to measure the expression levels of lncRNA PVT1 and miRNA-30a-5p.A one-way analysis of variance and the Tukey’s test were used to analyze the differences between multiple independent samples.Results YCHT significantly alleviated the pathological injury of the pancreas of SAP rats,such as edema,necrosis,hemorrhage,and inflammatory cell infiltration.Compared with the SO group,the SAP group had significant increases in the plasma levels of amylase and the inflammatory factors TNFαand IL-1β,and there were significant reductions in the plasma levels of amylase,TNFα,and IL-1βafter YCHT treatment(all P<0.05).Compared with the SO group,the SAP group had significant increases in LC-3II/LC-3I ratio and the protein expression of Beclin-1,XIAP,caspase-3,and NF-κB,and compared with the SAP group,the YCHT group had significant reductions in LC-3II/LC-3I ratio and the protein expression of Beclin-1,XIAP,and NF-κB(all P<0.05).Compared with the SO group,the SAP group
作者
项红
胡凤林
陶旭锋
齐心
张金楠
尚东
XIANG Hong;HU Fenglin;TAO Xufeng;QI Xin;ZHANG Jinnan;SHANG Dong(Clinical Laboratory of Integrative Medicine,The First Affiliated Hospital of Dalian Medical University,Dalian,Liaoning 116011,China;Third Department of General Surgery,The First Affiliated Hospital of Dalian Medical University,Dalian,Liaoning 116011,China;Institute(College)of Integrative Medicine,Dalian Medical University,Dalian,Liaoning 116044,China;School of Chemical Engineering,Dalian University of Technology,Dalian,Liaoning 116024,China)
出处
《临床肝胆病杂志》
CAS
北大核心
2021年第1期147-152,共6页
Journal of Clinical Hepatology
基金
国家自然科学基金面上项目(81873156)
国家重点研发计划“政府间国际科技创新合作/港澳台科技创新合作”重点专项项目(2018YFE0195200)。
