补肺益肾方调控NF-κB通路抑制香烟烟雾提取物诱导的MRC-5细胞炎症反应机制  

Mechanism of Bufei Yishen Formula regulating NF-κB pathway and inhibiting the inflammatory response of MRC-5 cells induced by cigarette smoke extract

作　　者：刘素晓 王洋[1,2] 芦晓帆 刘学芳[4] 冯素香[4] 郑万春 李亚[1,4] LIU Su-xiao;WANG Yang;LU Xiao-fan;LIU Xue-fang;FENG Su-xiang;ZHENG Wan-chun;LI Ya(Center Laboratory,Chinese Medicine Pharmacology(Respiratory)Laboratory,Henan Provincial Key Laboratory of Traditional Chinese Medicine for Prevention and Treatment of Respiratory Diseases,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;Respiratory Department,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;Respiratory Department,Henan Province Hospital of TCM,Zhengzhou 450002,China;Henan Key Laboratory of Chinese Medicine for Respiratory Disease,Henan University of Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学第一附属医院,中心实验室、中药药理(呼吸)实验室、河南省呼吸病防治中医药重点实验室,郑州450006 [2]河南中医药大学第一附属医院呼吸科,郑州450006 [3]河南省中医院呼吸科,郑州450006 [4]河南中医药大学河南省中医药防治呼吸病重点实验室,郑州450006

出　　处：《中华中医药杂志》2022年第10期5648-5652,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81302921,No.81673775,No.82074403);国家中医药管理局中医临床研究基地业务建设科研专项课题(No.JD2X2015157);河南省重点研发与推广专项(科技攻关)(No.182102310102)。

摘　　要：目的:观察补肺益肾方调控核因子-κB(NF-κB)通路对CSE诱导的人胚肺成纤维细胞(MRC-5)炎症反应的抑制作用。方法:正常培养和NF-κB p65 siRNA转染的MRC-5细胞经正常血清、5%CSE、5%CSE+补肺益肾方血清、5%CSE+氨茶碱血清处理24 h后,分别采用流式细胞术测细胞增殖、凋亡情况,ELISA方法测细胞因子白细胞介素(IL)-1β和肿瘤坏死因子α(TNF-α)水平,qRT-PCR检测IL-1β、TNF-α、NF-κB p65和IκBαmRNA表达水平,Western Blot检测p-NF-κB p65和p-IκBα蛋白表达水平。结果:CSE诱导可以显著降低正常和siRNA转染MRC-5细胞增殖率,增加凋亡率,提高IL-1β和TNF-α分泌及mRNA表达水平,以及NF-κB p65和IκBα基因及蛋白磷酸化水平(P<0.01);补肺益肾方处理可显著改善上述指标(P<0.01),且与NF-κB p65 siRNA有良好的协同作用。结论:补肺益肾方可以显著抑制CSE诱导的MRC-5细胞炎症反应,其机制可能与下调NF-κB信号通路有关。Objective: To observe the inhibition of Bufei Yishen Formula on inflammatory in MRC-5 induced by cigarette smoke extract(CSE) by regulating NF-κB pathway. Methods: Normally cultured MRC-5 and NF-κB p65 siRNA transfected MRC-5 were treated with normal serum, 5%CSE, 5%CSE+Bufei Yishen serum, 5%CSE+Aminophylline serum, respectively. After 24 h of treatment, cell proliferation and apoptosis were measured by flow cytometry, level of IL-1β and TNF-α was detected by ELISA, expressions of IL-1β, TNF-α, NF-κB p65 and IκBα mRNA were measured by qRT-PCR, and expressions of p-NF-κB p65 and p-IκBα protein were detected by Western Blot. Results: Induction of CSE could significantly reduce the rated of proliferation, increase the rate of apoptosis, increase the secretion and mRNA expression of IL-1β and TNF-α, and increase the mRNA and protein phosphorylation of NF-κB p65 and IκBα in normal and NF-κB p65 siRNA transfected MRC-5 cells(P<0.01). Treatment of Bufei Yishen Formula could significantly improve all the abovementioned changes(P<0.01), and it had a good synergistic effect with NF-κB p65 siRNA. Conclusion: Bufei Yishen Formula can significantly inhibit inflammation in MRC-5 cells induced by CSE, and the mechanism may be related to down-regulation of NF-κB pathway.

关 键 词：补肺益肾方 核因子-κB通路 香烟烟雾提取物 MRC-5细胞 炎症 

分 类 号：R285[医药卫生—中药学]