基于TLR4/NF-κB/NLRP3通路探讨参芪调肾方改善CSE诱导小鼠肺泡巨噬细胞炎症反应的作用机制  

To investigate the effect of Shenqi Tiaoshen Formula on CSE induced inflammatory response of MH⁃S cells based on TLR4/NF⁃κB/NLRP3 pathway

作　　者：王卉[1] 杨勤军 周凡超 杨程[2,3,4] 童佳兵 李泽庚[1,2,3,4,5] WANG Hui;YANG Qin-jun;ZHOU Fan-chao;YANG Cheng;TONG Jia-bing;LI Ze-geng(Anhui University of Traditional Chinese Medicine,Hefei 230038,China;The First Affiliated Hospital of Anhui University of Traditional Chinese Medicine,Hefei 230031,China;Institute of Respiratory Diseases,Anhui Academy of Chinese Medicine,Hefei 230031,China;Xin'an Key Laboratory of Medical Science,Ministry of Education,Hefei 230038,China;Key Laboratory of Application and Transformation of Traditional Chinese Medicine in Prevention and Treatment of Major Pulmonary Diseases,Hefei 230031,China)

机构地区：[1]安徽中医药大学,安徽合肥230038 [2]安徽中医药大学第一附属医院,安徽合肥230031 [3]安徽省中医药科学院中医呼吸病防治研究所,安徽合肥230031 [4]新安医学教育部重点实验室,安徽合肥230038 [5]中医药防治肺系重大疾病应用转化安徽省重点实验室,安徽合肥230031

出　　处：《海南医学院学报》2023年第17期1295-1301,共7页Journal of Hainan Medical University

基　　金：国家自然科学基金区域创新发展联合基金重点支持项目(U20A20398);安徽省科技厅临床医学研究转化专项(202204295107020045)。

摘　　要：目的:基于TLR4/NF-κB/NLRP3通路探讨参芪调肾方(SQTS)改善香烟烟雾提取物(Cigarette smoking extract,CSE)诱导小鼠肺泡巨噬细胞(MH-S)炎症反应的作用机制。方法:以MH-S细胞为研究对象,通过CCK-8法分析SQTS对MH-S细胞的安全性及药效浓度;随后MH-S分为空白组、模型组(8%CSE)和SQTS低、中、高剂量组(40、80、160 mg/L)。ELISA检测细胞上清TNF-α、IL-1β和IL-6含量;DCFH-DA荧光探针法检测细胞中ROS水平;Westernblot检测TLR4/NF-κB/NLRP3通路蛋白的表达,使用TLR4抑制剂TAK-242验证SQTS在TLR4/NF-κB/NLRP3通路上的作用。结果:与空白组比较,CSE组细胞活力降低,炎症因子TNF-α、IL-1β和IL-6的含量增加(P<0.05),ROS荧光表达水平显著升高(P<0.01),TLR4/NF-κB/NLRP3通路蛋白表达明显增加(P<0.05);与模型组比较,参芪调肾方各剂量组细胞活力升高,以上各指标表达水平均有所降低(P<0.05),且TAK-242各组细胞TLR4/NF-κB/NLRP3通路蛋白减少(P<0.05)。结论:参芪调肾方可减轻CSE诱导的MH-S细胞的炎症反应,其机制可能与抑制TLR4/NF-κB/NLRP3通路有关。Objective:To study the effects of Shenqi Tiaoshen Formula(SQTS)on the inflammatory response of MH⁃S cells induced by cigarette smoking extract(CSE)and its mechanism based on TLR4/NF⁃κB/NLRP3 pathway.Methods:MH⁃S cells were used as subjects to evaluate cell viability by CCK⁃8 method.The levels of TNF⁃α,IL⁃1βand IL⁃6 in the supernatant were detected by ELISA.ROS were detected by DCFH⁃DA fluorescence probe.Western blotting was used to detect the expres⁃sion of TLR4/NF⁃κB/NLRP3 pathway protein,and TAK⁃242,a TLR4 inhibitor,was used to verify the role of SQTS in the TLR4/NF⁃κB/NLRP3 pathway.Results:Compared with blank group,the cell survival rate of CSE group was decreased,and the contents of inflammatory cytokines TNF⁃α,IL⁃1βand IL⁃6 were increased(P<0.05).ROS fluorescence expression level was significantly increased(P<0.01).TLR4/NF⁃κB/NLRP3 pathway protein expression was significantly increased(P<0.05).Compared with model group,the survival rate of cells in SQTS groups was increased,and the expression levels of the above indexes were decreased(P<0.05).TLR4/NF⁃κB/NLRP3 pathway protein decreased in TAK⁃242 groups(P<0.05).Conclusion:SQTS can reduce the inflammatory response of MH⁃S cells induced by CSE by inhibiting TLR4/NF⁃κB/NLRP3 pathway.

关 键 词：参芪调肾方 CSE MH-S细胞 TLR4/NF-κB/NLRP3信号通路 炎症 

分 类 号：R285.5[医药卫生—中药学]