文献类型：期刊文献

中文题名：基于HMGB1/RAGE/NF-κB信号通路探讨大黄牡丹汤对急性胰腺炎肠损伤大鼠的干预作用及机制

英文题名：Dahuang Mudantang Alleviates Intestinal Injury in Rat Model of Acute Pancreatitis by Regulating HMGB1/RAGE/NF-κB Signaling Pathway

作者：文林林[1,2];汪永锋[1];白敏[1];汪湛东[1];宋冰[1,3];张延英[1,3];刘馨鸿[4];孙银凤[1];杨丹[1];王琼[1];赵泓彰[1];杨润泽[1]

第一作者：文林林

机构：[1]甘肃中医药大学,兰州730000;[2]庄浪县人民医院,甘肃平凉744600;[3]甘肃省实验动物行业技术中心,兰州730000;[4]甘肃农业大学,兰州730000

第一机构：甘肃中医药大学

年份：2023

卷号：29

期号：19

起止页码：1

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：国家自然科学基金项目(82160871);甘肃省中医药管理局重点项目(GZKZ-2021-10);甘肃省中医药管理局项目(GZK-2019-26);甘肃省自然科学基金项目(20JR5RA186)。

语种：中文

中文关键词：急性胰腺炎;大黄牡丹汤;肠损伤;炎症反应;高迁移率族蛋白B1(HMGB1)/晚期糖基化终产物(RAGE)/核转录因子-κB(NF-κB)信号通路

外文关键词：acute pancreatitis;Dahuang Mudantang;intestinal injury;inflammatory response;high-mobility group box 1(HMGB1)/receptor for advanced glycation endproduct(RAGE)/nuclear factor-kB(NF-kB)signaling pathway

摘要：目的:基于高迁移率族蛋白B1(HMGB1)/晚期糖基化终产物(RAGE)/核转录因子-κB(NF-κB)信号通路探讨大黄牡丹汤对急性胰腺炎肠损伤大鼠的干预作用及机制研究。方法:120只SPF级Wistar大鼠采用5%牛?胆酸钠逆行胰胆管注射制备急性胰腺炎(AP)肠损伤模型。随机分为正常组、模型组、大黄牡丹汤低、中、高剂量组(3.5、7、14 g·kg^(-1)),灌胃给药,奥曲肽组(1×10^(-5)g·kg^(-1)),皮下注射,每组20只,正常组和模型组大鼠给予等体积蒸馏水灌胃,于造模前1 h及造模后每隔12 h给药1次,造模后24 h收集样本。观察大鼠一般情况;生化法检测血清中淀粉酶(AMS)、C反应蛋白(CRP);酶联免疫吸附测定法(ELISA)检测结肠组织中的肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)及白细胞介素-6(IL-6)含量;苏木素-伊红(HE)染色观察胰腺和结肠组织病理形态变化;实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测结肠组织中HMGB1、RAGE、NF-κB抑制因子激酶(IKK)、核转录因子-κB抑制蛋白α(IκBα)、NF-κB mRNA及蛋白表达水平。结果:观察大鼠一般情况显示,与正常组比较,模型组大鼠出现一般生存状态不佳、扭体反应、大便次数减少及便质干等情况的改变;与模型组比较,各治疗组大鼠上述症状改善;与大黄牡丹汤低剂量组比较,大黄牡丹汤高剂量组症状改善最为明显。生化法检测血清AMS、CRP含量显示,与正常组比较,模型组大鼠AMS、CRP含量明显升高(P<0.05);与模型组比较,大黄牡丹汤组大鼠AMS、CRP含量降低(P<0.05);与大黄牡丹汤低剂量组比较,大黄牡丹汤高剂量组大鼠AMS、CRP含量明显降低(P<0.05)。ELISA检测结肠组织中TNF-α、IL-1β和IL-6含量显示,与正常组比较,模型组大鼠TNF-α、IL-1β、IL-6含量明显升高(P<0.05);与模型组比较,大黄牡丹汤组大鼠TNF-α、IL-1β、IL-6含量明显降低(P<0.05);与大黄牡丹汤低剂量组比较,大黄牡丹汤高剂量组和奥曲肽组大鼠TNF-α、IL-1β、IL-6含量明显降低(P<0.05)。HE染色结果显示,与正常组比较,模型组大鼠出现胰腺、结肠组织坏死、充血、结构不完整;与模型组比较,各治疗组大鼠的胰腺、结肠组织水肿、坏死程度明显改善;与大黄牡丹汤低剂量组比较,大黄牡丹汤高剂量组改善最为明显。Real-time PCR结果显示,与正常组比较,模型组大鼠HMGB1、RAGE、IKK、IκBα、NF-κBmRNA表达明显升高(P<0.05);与模型组比较,大黄牡丹汤各剂量组大鼠HMGB1、RAGE、IKK、IκBαmRNA表达水平明显降低(P<0.05);与大黄牡丹汤低剂量组比较,大黄牡丹汤中、高剂量组合奥曲肽组大鼠HMGB1、RAGE、IKK、IκBα、NF-κB mRNA表达水平降低,其中以大黄牡丹汤高剂量组表现最为明显(P<0.05)。Western blot结果与Real-time PCR分析结果趋势一致。结论:大黄牡丹汤对急性胰腺炎模型大鼠的一般情况、炎症指标及胰腺和结肠组织病理状态有较好的改善作用,其作用机制可能与抑制HMGB1/RAGE/NF-κB信号通路,减轻急性胰腺炎大鼠结肠组织细胞炎症反应有关。
Objective:To explore the mechanism of Dahuang Mudantang in alleviating the intestinal injury in the rat model of acute pancreatitis via the high-mobility group box 1(HMGB1)/receptor for advanced glycation endproduct(RAGE)/nuclear factor-κB(NF-κB)signaling pathway.Method:One hundred and twenty SPF-grade Wistar rats received retrograde injection of 5%sodium taurocholate into the biliopancreatic duct for the modeling of intestinal injury in acute pancreatitis.The rats were randomized into blank,model,low-,medium-,and high-dose(3.5,7,14 g·kg^(-1),administrated by gavage)Dahuang Mudantang,and octreotide(1×10-5 g·kg^(-1),subcutaneous injection)groups(n=20).The rats in blank and model groups received equal volume of distilled water by gavage.Drugs were administered 1 h before and every 12 h after modeling,and samples were collected 24 h after modeling.The general status of the rats was observed.The biochemical methods were employed to measure the levels of amylase(AMS)and C-reactive protein(CRP)in the serum.The enzyme-linked immunosorbent assay(ELISA)was employed to measure the levels of tumor necrosis factor(TNF)-α,interleukin(IL)-1β,and IL-6 in the colon tissue.The morphological changes of pancreatic and colon tissues were observed by hematoxylin-eosin(HE)staining.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)and Western blot were employed to measure the expression levels of HMGB1,RAGE,inhibitor of NF-κB kinase(IKK),and NF-κB suppressor proteinα(IκBα)in the colon tissue.Result:The rats in the model group showed poor general survival,writhing response,reduced frequency of defecation,and dry stool.The symptoms of rats in the model group were mitigated in each treatment group,and the high-dose Dahuang Mudantang showed the most significant effect.Compared with the normal group,the model group had elevated AMS and CRP levels(P<0.05),which were lowered by Dahuang Mudantang(P<0.05),especially that at the high dose(P<0.05).Compared with the normal group,the modeling elevated that levels of TNF-α,IL-1β,and IL-6(P<0.05).Such elevations were lowered by Dahuang Mudantang(P<0.05),and the high-dose group and the octreotide group showed better performance(P<0.05).The modeling caused necrotic,congested,and destructed pancreatic and colonic tissues,which were ameliorated by the drugs,especially high-dose Dahuang Mudantang.Compared with the normal group,the modeling up-regulated the mRNA levels of HMGB1,RAGE,IKK,IκBα,and NF-κB(P<0.05).Compared with the model group,Dahuang Mudantang and octreotide down-regulated the mRNA levels of HMGB1,RAGE,IKK,IκBα,and NF-κB(P<0.05),and the high-dose Dahuang Mudantang demonstrated the best performance(P<0.05).Western blot results showed a trend consistent with the results of Real-time PCR.Conclusion:Dahuang Mudantang can improved the general status,reduce inflammation,and alleviate histopathological changes in the pancreatic and colon tissues in the rat model of acute pancreatitis by inhibiting the HMGB1/RAGE/NF-κB signaling pathway.