基于Keap1/Nrf2/ARE信号通路探讨半夏泻心汤对慢性萎缩性胃炎大鼠的影响及作用机制  

Effect and Mechanism of Banxia Xiexintang on Rats with Chronic Atrophic Gastritis Based on Keap1/Nrf2/ARE Signaling Pathway

作　　者：石铖[1] 王茜[1] 刘宇[1] 郝蕾[1] 韩雪[1] 王彦刚 SHI Cheng;WANG Xi;LIU Yu;HAO Lei;HAN Xue;WANG Yan-gang(Hebei University of Chinese Medicine,Shijiazhuang 050200,China;The Third Affiliated Hospital of Beijing University of Chinese Medicine,Beijing 100018,China)

机构地区：[1]河北中医学院,石家庄050200 [2]北京中医药大学第三附属医院,北京100018

出　　处：《中国实验方剂学杂志》2021年第20期31-37,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北省高等学校科学技术研究项目(QN2016070)。

摘　　要：目的:探讨半夏泻心汤通过调控Kelch样环氧氯丙烷相关蛋白-1(Keap1)-核因子E2相关因子2(Nrf2)/抗氧化反应元件(ARE)信号通路防治慢性萎缩性胃炎的作用机制。方法:取SD大鼠,除12只正常组大鼠外,其余大鼠联合造模法复制大鼠慢性萎缩性胃炎(CAG)模型,造模成功后随即分为模型组,维酶素组(VG,60 mg·kg^(-1))及半夏泻心汤高、中、低剂量组(280,140,70 mg·kg^(-1)),分别相当于半夏泻心汤生药量28,14,7 g·kg^(-1)。正常组及模型组大鼠给予等体积的蒸馏水,各治疗组给予相应容积的药物灌胃。治疗12周后采用苏木素-伊红(HE)染色法观察CAG大鼠胃黏膜病理变化情况,蛋白免疫印迹法(Western blot)及实时荧光定量聚合酶链式反应(Real-time PCR)检测CAG大鼠胃黏膜中Nrf2,醌氧化还原酶-1(NQO1),谷胱甘肽-S-转移酶(GST)蛋白及mRNA的表达水平。结果:与正常组比较,模型组大鼠胃黏膜组织Nrf2,NQO1,GST蛋白及mRNA表达水平升高(P<0.05),胃黏膜萎缩,甚至肠化。与模型组比较,维酶素组及半夏泻心汤高、中剂量组Nrf2,NQO1,GST蛋白及mRNA表达水平降低(P<0.05),胃黏膜萎缩、肠化情况明显改善,且以高剂量组改善最为明显,但半夏泻心汤低剂量组作用不明显。结论:半夏泻心汤降低Nrf2的转录活性,关闭Nrf2信号通路,降低NQO1,GST的表达水平,实现正常的氧化-抗氧化平衡可能是其治疗CAG的作用机制之一。Objective: To explore the mechanism of Banxia Xiexintang(BXXX)in preventing and treating chronic atrophic gastritis(CAG)through Kelch-like ECH-associated protein 1(Keap1)/nuclear factor erythroid 2-related factor 2(Nrf2)/antioxidant response element(ARE)signaling pathway. Method: SD rats were divided into a normal group(n=12)and an experimental group for CAG model induction. The model rats were then randomly divided into a model group,a vatacoenayme(VG)group(60 mg·kg^(-1)),and high-(280 mg·kg^(-1)),medium-(140 mg·kg^(-1)),and low-dose(70 mg·kg^(-1))BXXX groups. The doses in the BXXX groups were equivalent to 28,14,and 7 g·kg^(-1) crude drugs. The rats in the normal group and the model group received distilled water at an equal volume,and those in the VG group and the BXXX groups were treated correspondingly by gavage. After 12 weeks of treatment,hematoxylin-eosin(HE)staining was carried out to observe pathological changes in the gastric mucosa of CAG rats. Western blot and real-time fluorescence-based quantitative PCR was used to detect the protein and mRNA expression levels of Nrf2,glutathione S-transferase(GST),and NAD(P)H:quinone oxidoreductase 1(NQO1)in the gastric mucosa of CAG rats. Result:Compared with the normal group,the model group showed increased protein and mRNA expression levels of Nrf2,NQO1,and GST in the gastric mucosa of the rats(P<0.05),atrophic gastric mucosa,and even intestinal metaplasia. The protein and mRNA expression levels of Nrf2,NQO1,and GST in the VG group and the highand medium-dose BXXX groups were lower than those in the model group(P<0.05),and gastric mucosa atrophy and intestinal metaplasia were significantly improved, especially in the high-dose BXXX group.However,the effect in the low-dose BXXX group was not significant. Conclusion: BXXX can blunt the transcriptional activity of Nrf2,shut down Nrf2 signaling pathway,and reduce the expression levels of NQO1 and GST to achieve normal oxidation-anti-oxidation balance,which may be one of its action mechanisms in the treatment o

关 键 词：半夏泻心汤 慢性萎缩性胃炎(CAG) Kelch样环氧氯丙烷相关蛋白-1(Keap1)-核因子E2相关因子2(Nrf2)/抗氧化反应元件(ARE)信号通路 NRF2 谷胱甘肽-S-转移酶(GST) 醌氧化还原酶-1(NQO1) 

分 类 号：R2-0[医药卫生—中医学] R22