二陈汤加味通过Jagged1/Notch1/Hes1信号通路对慢性阻塞性肺疾病的抗炎机制

Anti-inflammatory Mechanism of Modified Erchentang on Chronic Obstructive Pulmonary Disease Through Jagged1/Notch1/Hes1 Signaling Pathway

目的:观测二陈汤加味对慢性阻塞性肺疾病(COPD)大鼠肺组织中Jagged1/Notch1/Hes1信号通路中关键分子表达的影响,探讨二陈汤加味通过Jagged1/Notch1/Hes1信号通路对COPD抗炎的作用及分子机制。方法:将60只SD大鼠随机分为6组,分别为正常组、模型组、二陈汤加味低、中、高剂量(5、10、20·kg^(-1))和γ-分泌酶抑制剂(DAPT)组,每组10只。以烟熏联合气管滴注脂多糖(LPS)的方法制备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)测定各组血清中Notch1、可溶性细胞间黏附分子-1(sICAM-1)、白细胞活化黏附分子(ALCAM)和可溶性血管黏附分子-1(sVCAM-1)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Jagged1、Notch1和Hes1 mRNA表达,免疫组织化学(IHC)法检测大鼠肺组织中Jagged1、Notch1、Notch1胞内段(NICD1)和Hes1蛋白表达。结果:与正常组比较,模型组血清中Notch1、sICAM-1、ALCAM和sVCAM含量均显著升高(P<0.01);肺组织Jagged1、Notch1和Hes1 mRNA表达显著增加(P<0.01);Jagged1、Notch1、NICD1和Hes1蛋白的表达显著增强(P<0.01)。与模型组比较,二陈汤加味中、高剂量和DAPT组血清中Notch1、sICAM-1、ALCAM和sVCAM含量均明显减少(P<0.05,P<0.01);Jagged1、Notch1和Hes1 mRNA表达明显下调(P<0.05,P<0.01);Jagged1、Notch1、NICD1和Hes1蛋白表达均明显减弱(P<0.05,P<0.01)。结论:二陈汤加味能有效减弱COPD肺部炎症反应。其机制可能是通过下调Jagged1、Notch1、Hes1 mRNA表达,抑制Notch1、s ICAM-1、ALCAM和sVCAM释放,对抗炎症对COPD肺组织的损伤有关。

Objective:To observe the effect of modified Erchentang on the expression of key molecules in the Jagged1/Notch1/Hes1 signaling pathway in lung tissues of rats with chronic obstructive pulmonary disease(COPD)and explore its anti-inflammatory effect and molecular mechanism on COPD through the Jagged1/Notch1/Hes1 signaling pathway.Method:Sixty SD rats were randomly divided into normal group,model group,low-,medium-,and high-dose modified Erchentang groups(5,10,20 g·kg^(-1)),andγ-secretase inhibitor DAPT group(0.02 g·kg^(-1)),with 10 rats in each group.The COPD model was induced in rats by cigarette smoking combined with intratracheal instillation of lipopolysaccharide(LPS).Rats were treated with corresponding drugs by gavage,while those in the normal group and the model group were treated with the same amount of normal saline by gavage.The serum levels of Notch1,soluble intercellular adhesion molecule-1(sICAM-1),activated leukocyte cell adhesion molecule(ALCAM),and soluble vascular adhesion molecule-1(sVCAM-1)were detected by enzyme-linked immunosorbent assay(ELISA).The mRNA expression of Jagged1,Notch1,and Hes1 was detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).The protein expression of Jagged1,Notch1,Notch1 intracellular domain(NICD1),and Hes1 in lung tissues of rats was detected by immunohistochemistry(IHC).Result:Compared with the normal group,the model group showed increased serum content of Notch1,sICAM-1,ALCAM,and sVCAM-1(P<0.01),increased mRNA expression of Jagged1,Notch1,and Hes1 in lung tissues(P<0.01),and increased protein expression of Jagged1,Notch1,NICD1,and Hes1(P<0.01).Compared with the model group,the medium-and high-dose modified Erchentang groups and the DAPT group showed decreased serum content of Notch1,sICAM-1,ALCAM,and sVCAM-1(P<0.05,P<0.05),down-regulated mRNA expression of Jagged1,Notch1,and Hes1(P<0.05,P<0.01),and reduced protein expression of Jagged1,Notch1,NICD1,and Hes1(P<0.05,P<0.01).Conclusion:Modified Erchentang may inhibit the inflammatory response in the lung of COPD rats,and its mechanism may be related to the resistance of inflammatory injury in the lung by decreasing the mRNA expression of Jagged1,Notch1,and Hes1 and inhibiting the release of Notch1,sICAM-1,ALCAM,and sVCAM-1.