摘要
目的探讨黄芪建中汤通过Raf/MEK/ERK信号通路治疗大鼠脾胃虚寒型十二指肠溃疡(DU)的作用机制。方法将60只SD大鼠随机分为正常对照组、模型组、埃索美拉唑组(4.17 mg·kg^(-1))以及黄芪建中汤高、低剂量组(18.54、9.27 g·kg^(-1))。采用“苦寒泻下+劳倦过度”法联合阿司匹林、无水乙醇构建脾胃虚寒型DU大鼠模型。灌胃给药,每天1次,连续4 d。观察大鼠十二指肠黏膜损伤情况并计算溃疡指数和治疗指数;采用HE染色法观察十二指肠黏膜组织病理形态学改变;采用酶联免疫吸附试验检测十二指肠组织中前列腺素E2(PGE2)、白细胞介素10(IL-10)、肿瘤坏死因子α(TNF-α)水平;免疫荧光法检测十二指肠黏膜磷酸化丝氨酸/苏氨酸蛋白激酶(p-Raf)、磷酸化有丝分裂原活化蛋白激酶激酶1/2(p-MEK1/2)、磷酸化细胞外信号调节激酶1(p-ERK1)蛋白表达。结果与正常对照组比较,模型组大鼠的十二指肠溃疡指数显著升高(P<0.01);小肠绒毛脱落,隐窝脓肿,小肠绒毛长度和隐窝深度均显著降低(P<0.01);肠黏膜PGE2、IL-10含量均明显降低(P<0.01),TNF-α含量显著增加(P<0.01);肠黏膜p-Raf、p-MEK1/2和p-ERK1蛋白表达量均明显增加(P<0.01)。与模型组比较,黄芪建中汤各剂量组的大鼠十二指肠溃疡指数均明显降低(P<0.01);肠黏膜损伤得到明显改善,黏膜形态趋于完整,小肠绒毛长度和隐窝深度均明显增加(P<0.01);肠黏膜PGE2和IL-10含量均明显提高(P<0.01),TNF-α含量显著降低(P<0.01);肠黏膜p-Raf、p-MEK1/2和p-ERK1蛋白表达均显著下调(P<0.01)。结论黄芪建中汤对脾胃虚寒型DU大鼠具有治疗作用,其机制可能与调控PGE2、TNF-α、IL-10等炎症介质水平,抑制Raf/MEK/ERK信号通路活化有关。
Objective To investigate the effect and mechanism of Huangqi Jianzhong decoction(HQJZT)in the treatment of duodenal ulcer(DU)with spleen and stomach deficiency cold type syndrome in rats based on Raf/MEK/ERK signaling pathway.Methods Sixty SD rats were randomly divided into normal group,model group,esomeprazole group(4.17 mg·kg^(-1)),HQJZT low dose and high dose groups(18.54,9.27 g·kg^(-1)).DU model of spleen-stomach deficiency cold pattern was established by“bitter cold herbs+defatigation”method combined with aspirin and anhydrous ethanol.The injury of duodenal mucosa was observed with naked eyes,and the ulcer index(UI)and therapeutic index(TI)were determined.The histopathological changes of duodenal mucosa were observed with HE staining.The contents of PGE2,IL-10 and TNF-αin duodenal mucosa were detected with enzyme-linked immunosorbent assay.The expression of phosphorylated Raf,MEK1/2,and ERK1 protein in duodenal mucosa was detected with immunofluorescence.Results(1)Compared with the normal group,the UI of the DU model rats was significantly increased(P<0.01);the intestinal villi were shed and the crypt abscess was seen;the villi length and the crypt depth were both reduced(P<0.01).The contents of PGE2 and IL-10 in the intestinal mucosa were significantly reduced,and the content of TNF-αwas significantly increased(P<0.01).The expression levels of p-Raf,p-MEK1/2,and p-ERK1 protein in intestinal mucosa were significantly increased(P<0.01).(2)Compared with the model group,after treatment with HQJZT,the UI of rats was significantly reduced(P<0.01);the length of intestinal villi and the depth of crypts were significantly increased(P<0.01);the contents of intestinal mucosa PGE2 and IL-10 were significantly increased(P<0.01),TNF-αcontent was significantly reduced(P<0.01);the protein expression levels of p-Raf,p-MEK1/2,and p-ERK1 were significantly down-regulated(P<0.01).Conclusion HQJZT has a good therapeutic effect on duodenal ulcer in rats,and its mechanism may be related to the regulation of PGE2,TNF-αand IL-10,and the inhibition of the activation of Raf/MEK/ERK signaling pathway.
作者
宋厚盼
陈小娟
曾梅艳
陈新怡
杨焘
刘涛
吴海滨
仇婧玥
SONG Houpan;CHEN Xiaojuan;ZENG Meiyan;CHEN Xinyi;YANG Tao;LIU Tao;WU Haibin;QIU Jingyue(Hunan Provincial Key Laboratory of Diagnostic Research in Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208 Hunan,China;Faculty of Traditional Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208 Hunan,China;Shenzhen Traditional Chinese Medicine Hospital,Shenzhen 518033 Guangdong,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2021年第8期1093-1100,共8页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金项目(81703920)
中国博士后科学基金项目(2019M662790)
湖南省自然科学基金项目(2019JJ50442)
湖南省教育厅科学研究项目(19C1426)。
