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  

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 白敏[1] 汪湛东 宋冰 张延英 刘馨鸿[4] 孙银凤 杨丹 王琼 赵泓彰[1] 杨润泽[1] WEN Linlin;WANG Yongfeng;BAI Min;WANG Zhandong;SONG Bing;ZHANG Yanying;LIU Xinhong;SUN Yingfeng;YANG Dan;WANG Qiong;ZHAO Hongzhang;YANG Runze(Gansu University of Chinese Medicine,Lanzhou 730000,China;Zhuanglang People's Hospital,Pingliang 744600,China;Technology Center of Laboratory Animal Industry,Lanzhou 730000,China;Gansu Agricultural University,Lanzhou 730000,China)

机构地区：[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

出　　处：《中国实验方剂学杂志》2023年第19期1-8,共8页Chinese Journal of Experimental Traditional Medical Formulae

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)。

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�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 leve

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 

分 类 号：R2-0[医药卫生—中医学] R33R289R657.51R574