慢性阻塞性肺疾病大鼠模型CD8^(+)T细胞自噬水平研究及与其数量的相关性分析  

Autophagy level of CD8^(+)T cells in rat model of chronic obstructive pulmonary disease and correlation with its number

作　　者：陈宇鸣 于旭华 梁紫尧 李娜 陈思达[1,2] 申严 卓宋明[1,2] CHEN Yuming;YU Xuhua;LIANG Ziyao;LI Na;CHEN Sida;SHEN Yan;ZHUO Songming(College of Shenzhen Clinical Medical,Guangzhou University of Chinese Medicine,Shenzhen 518116,China;Shenzhen Longgang Central Hospital,Shenzhen 518116,China;The Second Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangzhou 510120,China)

机构地区：[1]广州中医药大学深圳临床医学院,深圳518116 [2]深圳市龙岗中心医院,深圳518116 [3]广州中医药大学第二附属医院,广州510120

出　　处：《中国免疫学杂志》2025年第2期295-303,共9页Chinese Journal of Immunology

基　　金：深圳市科技计划基础研究项目(JCYJ20160427100015608)。

摘　　要：目的:探究慢性阻塞性肺疾病(COPD)大鼠模型CD8^(+)T细胞自噬水平及与其数量的相关性,探讨自噬在COPD发病机制中的作用。方法:将36只6周龄SD雄性大鼠分为正常对照组、COPD组和COPD干预组,以烟熏方式建立COPD模型,腹腔注射给予3-甲基腺嘌呤(3-MA)干预。采用小动物肺功能仪检测大鼠肺功能,流式细胞术检测大鼠血液、肺组织CD8^(+)T细胞、CD8^(+)记忆T细胞、CD8^(+)效应T细胞比例,免疫磁珠分选血液CD8^(+)T细胞,Western blot检测CD8^(+)T细胞中LC3-Ⅱ/Ⅰ、Beclin-1、p62自噬蛋白表达水平,留取大鼠右肺中叶行病理检测。结果:COPD组大鼠肺组织炎症细胞浸润、肺功能下降较正常组大鼠明显。COPD组大鼠血液及肺组织CD8^(+)T细胞、CD8^(+)效应T细胞、CD8^(+)记忆T细胞比例均较正常对照组升高(P均<0.05),COPD组大鼠血液CD8^(+)T细胞自噬蛋白LC3-Ⅱ/Ⅰ、Beclin-1水平较正常对照组升高,p62水平较正常对照组降低(P<0.01)。COPD干预组大鼠肺组织中CD8^(+)T细胞、CD8^(+)效应T细胞、CD8^(+)记忆T细胞比例均较COPD组下降(P<0.05)。相关性分析显示,LC3-Ⅱ/Ⅰ表达水平与血液CD8^(+)T细胞比例呈正相关(r=0.667,P<0.01)Beclin-1表达水平与血液CD8^(+)T细胞、CD8^(+)效应T细胞比例呈正相关(r=0.505,P=0.021;r=0.428,P=0.037)。LC3-Ⅱ/Ⅰ表达水平与肺组织CD8^(+)T细胞、CD8^(+)效应T细胞比例呈正相关(r=0.474,P=0.019;r=0.549,P=0.006),Beclin-1表达水平与肺组织CD8^(+)效应T细胞比例呈正相关(r=0.458,P=0.025)。结论:CD8^(+)T细胞自噬水平与其数量相关,CD8^(+)T细胞自噬参与了COPD的慢性炎症过程,3-MA可抑制COPD炎症。Objective:To investigate the autophagy level of CD8^(+)T cells in rat model of chronic obstructive pulmonary disease(COPD)and correlation with its number,and to explore the role of autophagy in pathogenesis of COPD.Methods:Thirty-six 6-week-old male SD rats were divided into three groups,including normal control group,COPD group and COPD intervention group.COPD model was established by smoking,and intraperitoneal injection of 3-methyladenine(3-MA)was used for intervention.Lung function of rats was detected by small animal pulmonary function testing,and frequency of CD8^(+)T cells,CD8^(+)memory T cells,CD8^(+)effector T cells in blood and lung of rats were detected by flow cytometry.CD8^(+)T cells were sorted by immunomagnetic beads,and expressions of autophagy protein including LC3-Ⅱ/Ⅰ,Beclin-1 and p62 in CD8^(+)T cells were detected by Western blot.The middle lobe of right lung was collected for histological observation.Results:Infiltration of inflammatory cells and the decline of lung function in COPD group were more obvious than those in normal group.Frequency of CD8^(+)T cells,CD8^(+)effector T cells and CD8^(+)memory T cells in blood and lung of rats in COPD group were significantly higher than those in normal control group(all P<0.05).Level of autophagy protein,LC3-Ⅱ/Ⅰand Beclin-1 of CD8^(+)T cells in COPD group were significantly higher than that in normal control group,while level of p62 was lower than that in normal control group(P<0.01).CD8^(+)T cells,CD8^(+)effector T cells,CD8^(+)memory T cells in COPD intervention group were significantly lower than those in COPD group(P<0.05).Correlation analysis showed that LC3-Ⅱ/Ⅰexpression level was positively correlated with frequency of CD8^(+)T cells in blood(r=0.667,P<0.01).Expression level of Beclin-1 was positively correlated with frequency of CD8^(+)T cells and CD8^(+)effector T cells in blood(r=0.505,P=0.021;r=0.428,P=0.037).Expression of LC3-Ⅱ/Ⅰprotein was positively correlated with frequency of CD8^(+)T cells and CD8^(+)effector T cells

关 键 词：慢性阻塞性肺疾病 CD8^(+)T细胞 自噬 3-甲基腺嘌呤 

分 类 号：R563.3[医药卫生—呼吸系统]