六味补气方通过调控NLRP3/Caspase-1/GSDMD焦亡通路延缓大鼠慢性阻塞性肺疾病的发展  

Liuwei Buqi Formula delays progression of chronic obstructive pulmonary disease in rats by regulating the NLRP3/caspase-1/GSDMD pyroptosis pathway

作　　者：梅莉 张璐 吴迪 丁焕章 王新汝 张西安 卫宇航 李泽庚[2,3,4] 童佳兵 Li MEI;Lu ZHANG;Di WU;Huanzhang DING;Xinru WANG;Xian ZHANG;Yuhang WEI;Zegeng LI;Jiabing TONG(Graduate School of Anhui University of Traditional Chinese Medicine,Hefei 230012,China;First Affiliated Hospital of Anhui University of Traditional Chinese Medicine,Hefei 230031,China;Xin'an Institute of Medicine and Modernization of Traditional Chinese Medicine,Institute of Health Research,Hefei National Science Center,Hefei 230012,China;Anhui Provincial 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]安徽中医药大学研究生院,安徽合肥230012 [2]安徽中医药大学第一附属医院,安徽合肥230031 [3]合肥综合性国家科学中心大健康研究院,新安医学与中医药现代化研究所,安徽合肥230012 [4]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：《南方医科大学学报》2024年第11期2156-2162,共7页Journal of Southern Medical University

基　　金：国家自然科学基金(82374399);国家自然科学基金区域创新重点项目(U20A20398);安徽省教育厅重大项目(KJ2021ZD0063);合肥综合性国家科学中心大健康研究院新安医学与中医药现代化研究所“揭榜挂帅”重大突破项目(2023CXMMTCM005)。

摘　　要：目的基于NLRP3/Caspase-1/GSDMD的细胞焦亡通路,探讨六味补气方治疗慢性阻塞性肺疾病(COPD)模型大鼠的作用机制。方法将40只SD大鼠随机分为对照组、模型组、六味补气方组、六味补气方+MCC950组(n=10),除对照组外,其余各组大鼠采用烟熏联合脂多糖气管滴注以及激素注射法造模,建立COPD模型大鼠,并分别给予六味补气方药物灌胃及合并MCC950腹腔注射。观察各组大鼠的一般情况变化,分析大鼠肺组织病理改变、肺功能、肺泡灌洗液(BALF)吉姆萨染色总细胞及白细胞分类计数、血清中炎症因子IL-6、TNF-α、IL-18及NO水平;qRT-PCR检测大鼠肺组织中焦亡相关蛋白NLRP3、ASC、Caspase-1、GSDMD-N、IL-1β、IL-18的mRNA表达。结果与对照组相比,模型组大鼠的肺组织病理损伤较重,肺功能下降(P<0.01),BALF中总细胞数增多,白细胞分类计数均上调(P<0.01),IL-6、TNF-α、IL-18及NO表达增多(P<0.01),肺组织相关蛋白含量增多(P<0.01);与模型组相比,六味补气方组大鼠的肺组织病理、肺功能均有改善(P<0.05),BALF中总细胞数及白细胞分类计数减少(P<0.01),IL-6、TNF-α、IL-18及NO表达下降(P<0.01),肺组织相关焦亡蛋白含量减少(P<0.01);与六味补气方组大鼠相比,使用MCC950后的大鼠肺组织病理、肺功能有改善,但差异无统计学意义(P>0.05);BALF中总细胞数及白细胞分类计数减少(P<0.05),IL-6、TNF-α、IL-18及NO表达下降(P<0.05),肺组织相关焦亡蛋白含量较少(P<0.05)。结论六味补气方可能通过调节NLRP3/Caspase-1/GSDMD通路抑制COPD模型大鼠的肺组织焦亡,从而降低炎症反应,减轻肺部损伤,延缓疾病的发生发展。Objective To explore the therapeutic mechanism of Liuwei Buqi(LWBQ)Formula for chronic obstructive pulmonary disease(COPD)in rat models.Methods SD rat models of COPD established by cigarette smoking combined with intratracheal lipopolysaccharide(LPS)instillation and hormone injection were treated with LWBQ Formula by gavage with or without intraperitoneal injection of MCC950 for 3 weeks,starting at the 5th week of modeling.After the treatments,the rats were examined for lung pathologies,lung function,total cell count and white blood cell count in bronchoalveolar lavage fluid(BALF),and serum levels of IL-6,TNF-α,IL-18 and NO.The mRNA expressions of NLRP3,ASC,caspase-1,GSDMD-N,IL-1β,and IL-18 in the lung tissue were detected with qRT-PCR.Results Compared with the normal control rats,the COPD rat models had severe lung pathologies and showed significantly decreased lung function,increased total cell and leukocyte subset counts in BALF,and increased serum levels of IL-6,TNF-α,IL-18 and NO and mRNA expressions of pyroptosis-related proteins in the lung tissue.Treatment of the rat models with LWBQ Formula significantly improved lung pathology and lung function,reduced total cell and leukocyte counts in BALF,and decreased serum levels of the inflammatory factors and expressions of pyroptosis-related proteins in the lung tissue.The combined treatment with MCC950 further improved lung pathology and function in spite of a significant difference,but BALF cell counts,serum inflammatory factor levels and pulmonary expressions of pyroptosis-related proteins were all significantly reduced following the treatment.Conclusion LWBQ Formula can delay the progression of COPD in rats possibly by inhibiting lung tissue pyroptosis via regulating the NLRP3/caspase-1/GSDMD pathway to reduce inflammatory response and lung damage.

关 键 词：六味补气方 慢性阻塞性肺疾病 NLRP3/Caspase-1/GSDMD 细胞焦亡 MCC950 

分 类 号：R285.5[医药卫生—中药学]