基于TNF-α/TNFR1/RIPKs通路探讨二陈汤加味对COPD炎症的作用机制  

Mechanism of Modified Erchentang on COPD Inflammation Based on TNF-α/TNFR1/RIPKs Pathway

作　　者：陈壮 刘高阳 谢文英[1] 尚立芝[1] CHEN Zhuang;LIU Gaoyang;XIE Wenying;SHANG Lizhi(Henan University of Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学,郑州450046

出　　处：《中国实验方剂学杂志》2024年第9期40-47,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81573881);河南省科技攻关项目(182102311163)。

摘　　要：目的:基于肿瘤坏死因子-α(TNF-α)/肿瘤坏死因子受体1(TNFR1)/受体相互作用蛋白激酶(RIPKs)信号通路,研究二陈汤加味对慢性阻塞性肺疾病(COPD)大鼠模型炎症的影响,探讨其作用机制。方法:SD大鼠60只,随机分为正常组、模型组、二陈汤加味高、中、低剂量组、消咳喘组,每组10只。采用香烟烟雾联合脂多糖(LPS)方法制备COPD大鼠模型,二陈汤加味高、中、低剂量组分别以20、10、5 g·kg^(-1)·d^(-1)灌胃,消咳喘以3.5 mL·kg^(-1)·d^(-1)灌胃,正常组与模型组灌胃等量生理盐水,持续干预21 d。酶联免疫吸附测定法(ELISA)检测大鼠支气管肺泡灌洗液(BALF)中TNF-α、TNFR1的含量;实时荧光定量聚合酶链式反应(Real-time PCR)检测肺组织中受体相互作用蛋白激酶1(RIPK1)、受体相互作用蛋白激酶3(RIPK3)、混合系列激酶样结构域蛋白(MLKL)mRNA表达,蛋白免疫印迹法(Western blot)检测肺组织中RIPK1、RIPK3、MLKL蛋白表达,苏木素-伊红(HE)染色观察肺组织的病理变化。结果:与正常组比较,模型组大鼠BALF中TNF-α、TNFR1含量显著升高(P<0.01),肺组织中RIPK1、RIPK3、MLKL mRNA及蛋白表达水平显著升高(P<0.01)。与模型组比较,二陈汤加味高、中、低剂量组及消咳喘组大鼠BALF中TNF-α、TNFR1含量显著降低(P<0.01),肺组织中RIPK1、RIPK3、MLKL mRNA及蛋白表达水平有不同程度的降低(P<0.05,P<0.01)。结论:二陈汤加味能够有效改善COPD大鼠肺组织的炎症反应,其机制可能是通过抑制TNF-α/TNFR1/RIPKs信号通路。Objective:Based on tumor necrosis factor alpha(TNF-α)/tumor necrosis factor receptor 1(TNFR1)/receptor-interacting protein kinases(RIPKs)signaling pathway,this paper aims to study the effect of modified Erchentang on inflammation in rats with chronic obstructive pulmonary disease(COPD)and explore its mechanism of action.M ethod:A total of 60 SD rats were randomly divided into normal group,model group,high,medium,and low-dose groups(20,10,5 g·kg^(-1)·d^(-1))of modified Erchentang,and Xiaokechuan group(3.5 mL·kg^(-1)·d^(-1)),with 10 rats in each group.The COPD rat model was established by cigarette smoke combined with lipopolysaccharide(LPS).The normal group and model group were given the same amount of normal saline for 21 days by gavage administration.The contents of TNF-αand TNFR1 in bronchoalveolar lavage fluid(BALF)of rats were detected by enzyme-linked immunosorbent assay(ELISA).Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect mRNA expressions of RIPK1,RIPK3,and mixed lineage kinase domain-like(MLKL)in the lung tissue.The protein expressions of RIPK1,RIPK3,and MLKL in the lung tissue were detected by Western blot.The pathological changes in lung tissue were observed by hematoxylin-eosin(HE)staining.Result:Compared with the normal group,the contents of TNF-αand TNFR1 in BALF of the model group were significantly increased(P<0.01),and the mRNA and protein expression levels of RIPK1,RIPK3,and MLKL in the lung tissue were significantly increased(P<0.01).Compared with the model group,the contents of TNF-αand TNFR1 in BALF of high,medium,and low-dose groups of modified Erchentang and Xiaokechuan group were decreased(P<0.01).The mRNA and protein expression levels of RIPK1,RIPK3,and MLKL in the lung tissue were decreased to different degrees(P<0.05,P<0.01).Conclusion:Modified Erchentang can effectively improve the inflammatory response of lung tissue in COPD rats,and the mechanism may be by inhibiting the activation of the TNF-α/TNFR1/RIPKs signaling pathway.

关 键 词：二陈汤 慢性阻塞性肺疾病 炎症 肿瘤坏死因子-α(TNF-α)/肿瘤坏死因子受体1(TNFR1)/受体相互作用激酶(RIPKs)信号通路 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33