苗药“杆努尽烟”对慢性支气管炎大鼠TLR4-MyD88-NF-κB信号通路的影响  

Effect of Lysionotus pauciflorus Maxim.on Signaling Pathway of TLR4-MyD88-NF-κB in Lung Tissue of Rats with Chronic Bronchitis

作　　者：吴宁 黄雨霄 石雪 陈晋伦 彭凌峰 杨露露 徐红 孙见飞 刘华 WU Ning;HUANG Yuxiao;SHI Xue;CHEN Jinlun;PENG Lingfeng;YANG Lulu;XU Hong;SUN Jianfei;LIU Hua(Basic Medical Experimental Teaching Center,College of Basic Medicine,Guizhou Medical University,Guiyang 550025,Guizhou,China)

机构地区：[1]贵州医科大学基础医学院基础医学实验教学中心,贵州贵阳550025

出　　处：《贵州医科大学学报》2020年第11期1283-1288,共6页Journal of Guizhou Medical University

基　　金：贵州医科大学教学工程(201804021);国家级2018年大学生创新创业训练计划项目(201810660014)。

摘　　要：目的:探讨苗药“杆努尽烟”对慢性支气管炎(CB)大鼠Toll样受体4(TLR4)-髓样分化因子88(MyD88)-核转录因子-κB(NF-κB)信号通路的影响。方法:30只SD大鼠均分为正常对照组、模型组、低剂量“杆努尽烟”组(低剂量组,2 g/kg)、高剂量“杆努尽烟”组(高剂量组,8 g/kg)及桂龙咳喘宁阳性药物组(阳性对照组,1 g/kg),正常对照组于无烟环境中饲养,其余4组大鼠用烟熏联合脂多糖(LPS)复制CB大鼠模型,“杆努尽烟”组及阳性对照组给予相应药物28 d,处死大鼠取肺组织;显微镜下观察各组大鼠肺组织学变化、酶联免疫(ELISA)法检测肿瘤坏死因子-α(TNF-α)含量、实时荧光定量PCR(Real-time PCR)及蛋白免疫印迹(Western blot)法检测TLR4、MyD88、NF-κB p65 mRNA及蛋白表达水平。结果:与正常对照组比较,其余各组大鼠肺泡壁及细支气管结构受到破坏,有炎性细胞浸润,各组大鼠肺组织中TLR4、MyD88、NF-κB p65 mRNA及蛋白表达水平、TNF-α含量升高(P<0.05);与模型组比较,各给药组肺泡壁和细支气管损伤程度减轻,且各组大鼠肺组织中TLR4、MyD88、NF-κB p65 mRNA及蛋白表达水平、TNF-α含量降低(P<0.05),“杆努尽烟”高剂量组降低更显著(P<0.05)。结论:苗药“杆努尽烟”治疗CB的机制可能与下调TLR4-MyD88-NF-κB通路相关信号分子表达有关。Objective:To study the effect of Lysionotus pauciflorus Maxim.on expression of Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),and nuclear factor-κB(NF-κB)genes in the lung tissue homogenate of rats with chronic bronchitis(CB).Methods:Thirty SD rats were evenly divided into a normal control group,a model group,a low-dose(2 g/kg)Lysionotus pauciflorus Maxim.group,a high-dose(8 g/kg)Lysionotus pauciflorus Maxim.group,and a guilongkechuanning positive drug group(1 g/kg).The normal control group was raised in a smoke-free environment,other four groups were prepared through cigarette smoke and tracheal dripping with lipopolysaccharide(LPS)to replicate CB rat model.After giving corresponding drugs intragastrically for 28 days,the rats were killed and their lung tissue was excised.The pathological changes of lung tissue of each group was observed under microscopes,the content of tumor necrosis factor-α(TNF-α)in lung was detected by Enzyme-linked immunosorbent assay(ELISA),and the expression of TLR4,MyD88,NF-κB p65 mRNA and protein in the lung homogenate of each group was detected by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)and Western blot respectively.Results:Compared with the normal group,the lung tissue alveolar wall of the rats of the remaining groups was destroyed and inflammatory cell infiltration was detected.The content of TNF-αin plasma increased(P<0.05),and the mRNA and protein expressions of TLR4,MyD88,NF-κB p65 also increased(P<0.05).Compared with the model group,the alveolar wall and bronchiole structural integrity in each administration group were lighter,and the above indicators decreased(P<0.05),and the high-dose of Lysionotus pauciflorus Maxim.group showed a more significant decrease.Conclusion:Lysionotus pauciflorus Maxim.can inhibit the inflammatory response of CB effectively,which might be related to the inhibition of the expressions of the signal molecule genes involved in the TLR4-MyD88-NF-κB pathway.

关 键 词：支气管炎 慢性 大鼠 肿瘤坏死因子Α 杆努尽烟 TLR4/MyD88/NF-κB信号通路 肺组织 

分 类 号：R562.2[医药卫生—呼吸系统]