肌动蛋白相关蛋白2/3复合体在慢性阻塞性肺疾病小鼠肺泡巨噬细胞吞噬功能中的作用  

Effect of actin-related protein 2-3 complex on phagocytosis defect of alveolar macrophages in a mouse model of chronic obstructive pulmonary disease

作　　者：胡悦 霍少娟 曾晓丽[1] 包海荣[1] 刘晓菊[1] Hu Yue;Huo Shaojuan;Zeng Xiaoli;Bao Hairong;Liu Xiaoju(Department of Gerontal Respiratory Medicine,the First Hospital of Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学第一医院老年呼吸科,兰州730000

出　　处：《中华医学杂志》2019年第30期2355-2361,共7页National Medical Journal of China

基　　金：国家自然科学基金(81670033).

摘　　要：目的探讨肌动蛋白相关蛋白(Arp)2/3复合体在慢性阻塞性肺疾病(慢阻肺)小鼠肺泡巨噬细胞(AM)吞噬功能中的作用.方法40只小鼠按随机数字表法分为健康对照组、健康Arp2/3复合体抑制剂(CK666)组、慢阻肺组及慢阻肺CK666组各10只,慢阻肺组和慢阻肺CK666组采用香烟烟雾暴露法建立慢阻肺模型,其对照组无烟雾暴露.造模90d后,取各组小鼠肺组织分离AM.流式细胞术测AM吞噬异硫氰酸荧光素标记的大肠杆菌(FITC-E.coli)的能力,以平均荧光强度(MFI)和吞噬FITC-E.coli阳性细胞百分比(吞噬%)表示;蛋白印迹法测AM的Arp2和F-肌动蛋白表达;激光共聚焦显微镜测AM的Arp2、F-肌动蛋白、吞噬的FITC-E.coli平均吸光度及Arp2和F-肌动蛋白的共定位;扫描电镜观察AM吞噬时的形态.结果慢阻肺组AM的MFI和吞噬%均显著低于健康对照组[(4702±243)比(8684±234)和(32.21±1.66)%比(65.88±1.77)%,均P<0.01];慢阻肺CK666组[(3597±307)、(22.09±1.89)%]、健康CK666组[(7446±236)、(50.09±1.64)%]均显著低于各自的对照组(均P<0.01).慢阻肺组AM的Arp2和F-肌动蛋白相对表达量均显著低于健康对照组(0.508±0.025比0.813±0.040和0.462±0.029比0.720±0.039,均P<0.01);慢阻肺CK666组(0.265±0.014)、健康CK666组(0.637±0.032)F-肌动蛋白均显著低于各自对照组(均P<0.01).慢阻肺组AM的Arp2蛋白、F-肌动蛋白、FITC-E.coli平均吸光度均显著低于健康对照组(34.43±0.56比142.83±1.90、61.59±0.70比145.93±3.05、41.49±0.33比189.17±2.60,均P<0.01);慢阻肺CK666组(37.73±1.04、28.84±2.95)和健康CK666组(137.07±1.35、157.46±1.00)F-肌动蛋白、FITC-E.coli平均吸光度均显著低于各自对照组(均P<0.01).慢阻肺组孟德斯共定位系数(MOC)显著低于健康对照组(0.395±0.014比0.880±0.002,P<0.01),慢阻肺CK666组(0.297±0.006)、健康CK666组(0.737±0.031)均显著低于各自对照组(均P<0.01).健康对照组AM伸出密集且长的丝状伪足,细胞表面突起、皱褶�Objective To investigate the role of actin-related protein 2-3 complex (Arp2/3) complex on phagocytosis of alveolar macrophages (AMs) in a mouse model of chronic obstructive pulmonary (COPD). Methods Forty mice were randomly divided into healthy control group, healthy Arp2/3 complex inhibitor (CK666) group, COPD group and COPD CK666 group with 10 mice in each group. COPD group and COPD CK666 group were established by cigarette smoke exposure, and the control group had no smoke exposure. After 90 days of molding, AMs were isolated from lung tissue of mice in each group. Mean fluorescence intensity (MFI) and the positive percent of AMs engulfing fluorescein isothiocyanate-labeled Escherchina coli (FITC-E.coli)(AM%) were detected by flow cytometry. Western blot was applied to detect protein. Laser scanning confocal microscopy was used to measure the mean optical density of Arp2, F-actin and engulfed FITC-E. coli and quantify the colocalization of Arp2 and F-actin by a Manders′ overlap coefficient. Scanning electron microscopy was used to observe the ultrastructure of AM phagocytizing FITC-E.coli. Results Phagocytosis of AM: MFI and AM% in the COPD group were significantly decreased than those in the healthy control group[(4 702±243),(8 684±234) and (32.21±1.66)%,(65.88±1.77)%, all P<0.01]. MFI and AM% in the COPD CK666 group [(3 597±307),(22.09±1.89)%] and in the healthy CK666 group [(7 446±236),(50.09±1.64)%] were decreased compared to those in their respective control groups (all P<0.01). The expressions of protein of Arp2 and F-actin in the COPD group were significantly decreased than those in the healthy control group (0.508±0.025, 0.813±0.040 and 0.462±0.029, 0.720±0.039)(all P<0.01). The F-actin in the COPD CK666 group (0.265±0.014) and in the healthy CK666 group (0.637±0.032) were significantly decreased compared to those in their respective control groups (all P<0.01). The mean optical density of Arp2, F-actin and FITC-E.coli in the COPD group were significantly decreased compared to those in

关 键 词：肺疾病 慢性阻塞性 巨噬细胞 肺泡 肌动蛋白相关蛋白2-3复合物 吞噬作用 

分 类 号：R563.9[医药卫生—呼吸系统]