摘要
目的:观察二陈汤加味对慢性阻塞性肺疾病(COPD)大鼠肺Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核转录因子-κB(NF-κB)通路及相关分子的影响,探讨二陈汤加味对COPD抗炎作用的分子机制。方法:将40只SD大鼠随机平均分为正常组、模型组、二陈汤加味组和EVP4593(NF-κB抑制剂)组。以香烟烟雾联合气管滴注脂多糖(LPS)方法制备COPD大鼠模型。二陈汤加味组以10 g·kg-1灌胃(ig),EVP4593组皮下注射1 mg·kg-1,正常组、模型组ig等量生理盐水,连续干预14 d。酶联免疫吸附测定法(ELISA)检测血清高迁移率族蛋白B1(HMGB1),趋化因子(CXCL)-2,CXCL-3和单核细胞趋化蛋白-1(MCP-1)水平,实时荧光定量聚合酶链式反应(Real-time PCR),蛋白免疫印迹法(Western blot)分别检测肺匀浆中TLR4,MyD88,NF-κB p65 mRNA及蛋白表达水平,免疫组化(IHC)检测TLR4,MyD88,磷酸化NF-κB p65(p-NF-κB p65)在大鼠肺组织中的定位表达。结果:与正常组比较,模型组肺匀浆中TLR4,MyD88,NF-κB p65 mRNA及蛋白表达均显著增加(P<0.01),HMGB1,CXCL-2,CXCL-3和MCP-1含量显著升高(P<0.01)。与模型组比较,二陈汤加味组TLR4,MyD88,NF-κB p65mRNA及其蛋白表达均显著减弱(P<0.05,P<0.01),HMGB1,CXCL-2,CXCL-3和MCP-1含量显著降低(P<0.05,P<0.01)。结论:二陈汤加味对COPD有抗炎作用,其机制可能与抑制TLR4/MyD88/NF-κB通路相关信号分子基因的表达,以及减少HMGB1,CXCL-2,CXCL-3和MCP-1的释放有关。
Objective:To study the effect of modified Erchentang on expressions of Toll-like receptor 4(TLR4),myeloid differentiation factor(MyD88)and nuclear factor-κB(NF-κB)genes in the lung tissue homogenate of rats with chronic obstructive pulmonary disease(COPD).Method:Forty SD rats were randomly divided into normal group,model group,modified Erchentang group and EVP4593(NF-κB inhibitor)group.Ra COPD models were prepared through cigarette smoke and tracheal dripping with lipopolysaccharide(LPS).After the modeling,normal and model groups were intragastrically given normal saline solution,EVP4593 group was given EVP4593(1 mg·kg-1)through subcutaneous injection,and modified Erchentang group was given corresponding herbal drugs intragastrically(10 g·kg-1)for 14 days.The levels of high mobility group box 1(HMGB1),chemokines CXCL-2,CXCL-3 and monocyte chemoattractant protein-1(MCP-1)in rats serum were detected by enzyme-linked immunosorbent assay in rats serum.The expressions of Toll-like receptors 4(TLR4),myeloid differentiation factor(MyD88)and nuclear factor-κB p65(NF-κB p65)mRNA were detected by Realtime fluorescence quantitative PCR(Real-time PCR)method.Western blot were used to detect the levels of TLR4,MyD88,NF-κB p65 and p-NF-κB p65 protein.Immunohistochemistry(IHC)method was used to detect the localization and expressions of TLR4,MyD88 and p-NF-κB p65 protein in the lung tissue.Result:The mRNA and protein expressions of TLR4,My D88 and NF-κB p65 were increased significantly(P<0.01)in model group compared with normal group.The levels of HMGB1,CXCL-2,CXCL-3 and MCP-1 in the model group were significantly higher than those in the normal group(P<0.01).Compared with model group,expressions of TLR4,MyD88 and NF-κB p65 mRNA and protein were decreased significantly(P<0.05)in modified Erchentang.Conclusion:Modified Erchentang may inhibit the inflammatory response of COPD effectively.The mechanism may be related to the inhibition of the expressions of the signal molecule genes involved in the TLR4/MyD88/NF-κB path
作者
尚立芝
季书
王国强
陈晓辉
谢文英
李耀洋
张光远
石龙涛
徐莉莉
SHANG Li-zhi;JI Shu;WANG Guo-qiang;CHEN Xiao-hui;XIE Wen-ying;LI Yao-yang;ZHANG Guang-yuan;SHI Long-tao;XU Li-li(Henan University of Chinese Medicine,Zhengzhou 450046,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2019年第23期65-72,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(81573881)
河南省高等学校重点科研项目(15A360030)
河南省科重点技攻关(182102311163)
国家级大学生创新创业训练计划项目(S201810471013)
河南中医药大学生创新学习项目(CXXM[2017]0252)
关键词
慢性阻塞性肺疾病
二陈汤
Toll样受体4
核转录因子-ΚB
P65
单核细胞趋化蛋白-1
chronic obstructive pulmonary disease(COPD)
Erchentang
Toll-like receptors 4(TLR4)
nuclear factor-kappa B p65(NF-κB p65)
monocyte chemoattractantprotein-1(MCP-1)
