摘要
目的:探讨二陈汤加味对慢性阻塞性肺疾病(COPD)大鼠肺组织中肝素结合因子(Midkine)/跨膜受体蛋白(Notch2)/Hey1信号通路中的影响及其抗炎的分子机制。方法:60只SD大鼠随机分为正常组、模型组、二陈汤加味低、中、高剂量组(5、10、20 g·kg^(-1)·d^(-1))和γ-分泌酶抑制剂组(Notch1通路抑制剂,DAPT),每组10只。以烟熏联合脂多糖(LPS)法制备COPD大鼠模型。二陈汤加味各干预组灌胃(ig)给药;DAPT组ig DAPT(0.02 g·kg^(-1));正常组及模型组ig等体积生理盐水。采用酶联免疫吸附测定法(ELISA)检测支气管肺泡灌洗液(BALF)中Midkine、中性粒细胞趋化因子-1(CINC-1)、大鼠巨噬细胞来源趋化因子(MDC)、大鼠CXC趋化因子5(CXCL5)、中性粒细胞弹性蛋白酶(NE)和核转录因子-κB(NF-κB)p65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Midkine、Notch2和Hey1 mRNA表达;免疫组化(IHC)检测大鼠肺组织中Midkine、Notch2和Hey1蛋白表达。结果:与正常组比较,模型组BALF中Midkine、CINC-1、MDC、CXCL5、NE和NF-κB p65含量均显著升高(P<0.01);肺组织Midkine、Notch2和Hey1 mRNA和蛋白表达均显著增加(P<0.01)。与模型组比较,二陈汤加味中、高剂量组和DAPT组BALF中Midkine、CINC-1、MDC、CXCL5和NF-κB p65含量均显著减少(P<0.01);Midkine、Notch2和Hey1 mRNA表达显著下降(P<0.01)。结论:二陈汤加味对COPD有抗炎作用。其机制可能是通过下调Midkine、Notch2和Hey1 mRNA表达,抑制Midkine、CINC-1、MDC、CXCL5释放有关。
Objective:To investigate the molecular mechanism of the anti-inflammatory effect of Erchentang in the lung tissue of the rat model of chronic obstructive pulmonary disease(COPD)via the heparinbinding factor(Midkine)/transmembrane receptor protein(Notch2)/Hey1 signaling pathway.Method:Sixty SD rats were randomized into normal group,model group,modified Erchentang(5,10,20 g·kg^(-1)·d^(-1))groups,and Notch1 pathway inhibitor(γ-secretase inhibitor,DAPT,0.02 g·kg^(-1))group,with 10 rats in each group.The rat model of COPD was established by cigarette smoke combined with lipopolysaccharide(LPS).After the modeling,the rats were administrated with corresponding drugs by gavage,and those in the normal and model groups were administrated with normal saline by gavage for 21 days.The levels of Midkine,cytokine-induced neutrophil chemoattractant-1(CINC-1),macrophage-derived chemokine(MDC),chemokine ligand 5(CXCL5),neutrophil elastase(NE),and nuclear factor-kappa B(NF-κB)p65 in bronchoalveolar lavage fluid(BALF)were determined by enzyme-linked immunosorbent assay(ELISA).Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)and immunohistochemistry were respectively employed to determine the mRNA and protein levels of Midkine,Notch2,and Hey1 in the lung tissue.Result:Compared with the normal group,the modeling increased the levels of Midkine,CINC-1,MDC,CXCL5,NE,and NF-κB p65 in BALF(P<0.01)and up-regulated the mRNA and protein levels of Midkine,Notch2,and Hey1 in the lung tissue(P<0.01).Compared with the model group,medium-and high-dose modified Erchentang and DAPT lowered the levels of Midkine,CINC-1,MDC,CXCL5,and NF-κB p65 in BALF(P<0.01)and downregulated the mRNA levels of Midkine,Notch2,and Hey1(P<0.01).Conclusion:Modified Erchentang may inhibit the inflammation in COPD rats by down-regulating the expression of Midkine,Notch2,and Hey1 and reducing the content of Midkine,CINC-1,MDC,and CXCL5.
作者
胡文豪
尚立芝
谢文英
李耀洋
刘高阳
陈壮
HU Wenhao;SHANG Lizhi;XIE Wenying;LI Yaoyang;LIU Gaoyang;CHEN Zhuang(Henan University of Chinese Medicine,Zhengzhou 450046,China;Henan Provincial Second People's Hospital,Zhengzhou 451191,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第16期12-21,共10页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(81573881)
河南省科技攻关项目(182102311163)。
作者简介
第一作者:胡文豪,硕士,主治医师,从事中医药临床研究,E-mail:huwenhao1108@126.com;通信作者:尚立芝,硕士,教授,硕士生导师,从事中医药作用机制研究,E-mail:lzshang2014@163.com;通信作者:谢文英,硕士,教授,硕士生导师,从事中医药治疗肺系疾病的临床研究,E-mail:xiewenying1963@163.com。
