STAT1对肺结核大鼠巨噬细胞凋亡及MAPK/NF-κB信号通路的机制研究  

Study on the mechanism of STAT1 in apoptosis of rat macrophages and MAPK/NF-κB signaling pathway in pulmonary tuberculosis

作　　者：柯玲玲 王敏 高洁 孙建斌 KE Ling-ling;WANG Ming;GAO Jie;SUN Jian-bin(Laboratory Department of Wuhan Third Hospital,Wuhan,Hubei Province,430060,China)

机构地区：[1]武汉市第三医院检验科,湖北武汉430060

出　　处：《解剖学研究》2024年第2期115-122,共8页Anatomy Research

基　　金：武汉市公共卫生及卫生政策科研项目(WG14C20)。

摘　　要：目的探讨信号转导和转录激活因子1(STAT1)对肺结核大鼠巨噬细胞凋亡及MAPK/NF-κB信号通路的影响。方法将40只大鼠按照随机数字表达分为4组,即Control组、PTB组、STAT1-ASO组和STAT1-ASO+Anisomycin组,每组10只。计数肺组织结核分枝杆菌菌落;收集支气管肺泡巨噬细胞收集,流式细胞仪检测肺组织巨噬细胞细胞凋亡;HE染色检测肺组织病理学变化;ELISA检测血清中IL-6、IL-1β、TNF-α水平比较;检测肺组织中SOD、MDA水平;蛋白质印迹法检测创面组织中p38 MAPK、p-p38 MAPK、p65 NF-κB、p-p65 NF-κB蛋白表达。结果与Control组比较,PTB组大鼠肺组织结核分枝杆菌菌落数(9.12±0.81)、巨噬细胞凋亡率[(51.27±4.16)%]、血清中IL-6(215.42±15.33)、IL-1β(73.62±6.04)、TNF-α(81.24±5.79)水平、肺组织中MDA含量(9.54±1.72)及p-p38 MAPK/p38 MAPK(0.92±0.12)、p-p65 NF-κB/p65NF-κB比值(0.87±0.10)均明显增加,肺组织中SOD活性(31.27±2.85)明显降低(P<0.05);与PTB组比较,STAT1-ASO组大鼠肺组织结核分枝杆菌菌落数(3.87±0.40)、巨噬细胞凋亡率(8.91±0.43)、血清中IL-6(40.91±3.46)、IL-1β(21.54±1.79)、TNF-α水平(20.35±1.87)、肺组织中MDA含量(2.69±0.72)及p-p38 MAPK/p38 MAPK(0.45±0.07)、p-p65 NF-κB/p65 NF-κB比值(0.39±0.06)明显减少,肺组织中SOD活性(72.15±6.81)明显升高(P<0.05);与STAT1-ASO组比较,STAT1-ASO+Anisomycin组大鼠肺组织结核分枝杆菌菌落数(8.75±0.74)、巨噬细胞凋亡率(42.86±3.75)、血清中IL-6(192.11±13.61)、IL-1β(67.33±5.24)、TNF-α水平(75.26±5.11)、肺组织中MDA含量(9.01±1.65)及p-p38 MAPK/p38 MAPK(0.87±0.10)、p-p65 NF-κB/p65NF-κB比值(0.80±0.09)均明显增加,肺组织中SOD活性(36.49±3.01)明显降低(P<0.05)。结论抑制STAT1活化可抑制肺结核大鼠巨噬细胞凋亡,其作用机制可能和抑制MAPK/NF-κB信号通路活化有关。Objective To explore the effects of STAT1 on apoptosis of rat macrophages and MAPK/NF-κBsignaling pathway in pulmonary tuberculosis.Methods Forty rats were randomly divided into four groups,including the control group,PTB group,STAT1-ASO group,and STAT1-ASO+Anisomycin group,with 10 rats in eachgroup.The number of Mycobacterium tuberculosis colonies in lung tissues was counted.Bronchoalveolar lavage macrophages were collected,and apoptosis of lung tissue macrophages was detected by flow cytometry.Lung tissue histopathological changes were examined by HE staining.The levels of IL-6,IL-1β,and TNF-αin serum were measuredby ELISA.The levels of SOD and MDA in lung tissues were determined.Protein expression of p38 MAPK,p-p38MAPK,p65 NF-κB,and p-p65 NF-κB in wound tissue was detected by Western blotting.Results Compared with the control group,the TB group showed significant increases in the number of Mycobacterium tuberculosis colonies(9.12±0.81),macrophage apoptosis rate(51.27±4.16),serum IL-6(215.42±15.33),IL-1β(73.62±6.04),TNF-α(81.24±5.79)levels,lung tissue MDA content(9.54±1.72),and p-p38 MAPK/p38 MAPK ratio(0.92±0.12),aswell as p-p65 NF-κB/p65 NF-κB ratio(0.87±0.10),while pulmonary SOD activity(31.27±2.85)was significantlydecreased(P<0.05).Compared with the TB group,the STAT1-ASO group showed significant decreases in the number of M.tuberculosis colonies(3.87±0.40),macrophage apoptosis rate(8.91±0.43),serum IL-6(40.91±3.46),IL-1β(21.54±1.79),TNF-α(20.35±1.87)levels,lung tissue MDA content(2.69±0.72),and p-p38 MAPK/p38MAPK ratio(0.45±0.07),as well as p-p65 NF-κB/p65 NF-κB ratio(0.39±0.06),while pulmonary SOD activity(72.15±6.81)was significantly increased(P<0.05).Compared with the STAT1-ASO group,the STAT1-ASO+Anisomycin group showed significant increases in the number of Mycobacterium tuberculosis colonies(8.75±0.74),macrophage apoptosis rate(42.86±3.75),serum IL-6(192.11±13.61),IL-1β(67.33±5.24),TNF-α(75.26±5.11)levels,lung tissue MDA content(9.01±1.65),and p-p38 MAPK/p38 MAP

关 键 词：肺结核 信号转导和转录激活因子1 巨噬细胞凋亡 丝裂原活化蛋白激酶/核转录因子κB(MAPK/NF-κB)信号通路 

分 类 号：R521[医药卫生—内科学]