血红素氧化酶1调控自噬对脓肿分枝杆菌胞内生长的影响  

Effects of heme oxygenase-1 on autophagy regulation and intracellular growth of Mycobacterium abscessus

作　　者：刘爱凤[1] 罗海军[1] 谢仁峰 马小华[1] LIU Aifeng;LUO Haijun;XIE Renfeng;MA Xiaohua(Changsha Central Hospital,Changsha 410004,China)

机构地区：[1]长沙市中心医院,长沙410004

出　　处：《中国免疫学杂志》2025年第4期873-878,共6页Chinese Journal of Immunology

基　　金：湖南省卫生健康委课题(20201287);湖南省自然科学基金项目(2021JJ70130,2025JJ80543)。

摘　　要：目的:探讨血红素氧化酶1(HO-1)调控自噬对脓肿分枝杆菌(M.abs)胞内生长的影响。方法:M.abs标准株ATCC19977按照指定的感染复数(MOI)刺激小鼠RAW264.7巨噬细胞特定时间,Western blot检测HO-1蛋白表达水平和自噬相关蛋白Atg5、LC3Ⅱ、p62蛋白表达水平。采用HO-1特异性诱导剂CoPP和酶活性抑制剂SnPP预处理巨噬细胞12 h,加入M.abs共孵育2 h,阿米卡星去除胞外菌后继续培养至指定时间,CCK-8法检测细胞活性;Western blot、LysoTracker Red检测HO-1蛋白与自噬相关蛋白的调节关系;菌落计数、ELISA分别检测细胞内细菌存活情况和TNF-α分泌水平。结果:与对照组相比,M.abs能诱导HO-1和Atg5、LC3Ⅱ、p62蛋白表达水平升高;HO-1过表达和抑制能有效调控M.abs诱导的Atg5、LC3Ⅱ和p62表达。LysoTracker Red、菌落计数和ELISA结果显示,SnPP促进细胞内溶酶体增加,显著抑制M.abs胞内繁殖并减少TNF-α分泌。结论:抑制HO-1能增强M.abs诱导的自噬流,减少M.abs胞内生长,为研发以抑制HO-1为靶点的靶向药物提供科学依据。Objective:To investigate the role of heme oxygenase-1(HO-1)regulating autophagy and its impact on the intracellular growth of Mycobacterium abscessus(M.abs).Methods:The standard strain of M.abs ATCC19977 was used to stimulate mouse RAW264.7 macrophages with a specified multiple of infection(MOI)for a specified time.Western blot analysis was performed to detect the expression levels of HO-1 protein and autophagy related proteins Atg5,LC3Ⅱand p62.Macrophages were pretreated with HO-1 specific inducer CoPP and enzyme activity inhibitor SnPP for 12 h,and incubated with M.abs for 2 h.Amikacin removed extracellular bacteria and continued to culture until the specified time,and cell activity was detected by CCK-8 method.Western blot and LysoTracker Red were used to detect the regulatory relationship between HO-1 protein and autophagy related proteins.Bacterial survival and TNF-αsecretion levels were detected by colony count and ELISA.Results:Compared with the control group,M.abs could induce increased expression of HO-1,Atg5,LC3Ⅱand p62 proteins.Over-expression and inhibition of HO-1 were found to effectively regulate the expressions of Atg5,LC3Ⅱand p62 induced by M.abs.LysoTracker Red,colony count,and ELISA results demonstrated that SnPP promoted the increasement of intracellular lysosomes,significantly inhibited M.abs intracellular proliferation,and reduced TNF-αsecretion.Conclusion:Inhibition of HO-1 can enhance the autophagy flow induced by M.abs and reduce the intracellular growth of M.abs,which provides scientific basis for the development of targeted drugs targeting HO-1.

关 键 词：脓肿分枝杆菌 血红素氧化酶1 自噬 菌落计数 

分 类 号：R378[医药卫生—病原生物学]