环鸟苷酸腺苷酸合成酶-环鸟苷酸腺苷酸-干扰素基因刺激分子信号通路在结核性胸膜炎大鼠中的作用  

Role of cyclic guanosine adenylate synthetase-cyclic guanosine monophosphate adenosine monophosphate-stimulator of interferon gene signal pathway in rats with tuberculous pleurisy

作　　者：黄健[1,2] 于建 李明瑛[1,2] 韩伟[1,2] 崔秀琴 HUANG Jian;YU Jian;LI Mingying;HAN Wei;CUI Xiuqin(Department of Tuberculosis,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China;Key Laboratory of Tuberculosis of Xinxiang City,Weihui 453100,Henan Province,China;Department of Pathology,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China)

机构地区：[1]新乡医学院第一附属医院结核内科,河南卫辉453100 [2]新乡市结核病学重点实验室,河南卫辉453100 [3]新乡医学院第一附属医院病理科,河南卫辉453100

出　　处：《新乡医学院学报》2023年第4期313-318,323,共7页Journal of Xinxiang Medical University

摘　　要：目的探讨环鸟苷酸腺苷酸合成酶(cGAS)-环鸟苷酸腺苷酸(cGAMP)-干扰素基因刺激分子(STING)信号通路在结核性胸膜炎大鼠中的作用。方法30只6周龄无特定病原级Sprague Dawley雄性大鼠适应性饲养5 d后随机分为对照组、模型组和cGAS抑制剂组,每组10只。3组大鼠适应性饲养5 d后于右侧腹股沟内侧皮内注射100μL卡介苗(BCG)悬液(0.06 mg);接种5周后,模型组和cGAS抑制剂组大鼠于右侧肋弓角顶点注射1 mL结核分枝杆菌H37RV悬液(0.03 mg),建立结核性胸膜炎大鼠模型;对照组大鼠仅接种BCG,不注射结核分枝杆菌H37RV悬液。cGAS抑制剂组大鼠自造模第2天尾静脉注射cGAS抑制剂RU.521(600μg·L^(-1))200μL,每日1次,连续7 d;对照组和模型组大鼠尾静脉注射等体积的生理盐水。第8天脱颈处死大鼠,立刻从大鼠剑突侧处沿胸骨剪开皮肤和胸骨,暴露胸腔和纵隔,观察大鼠胸腔积液的分布情况,收集、记录胸腔积液量;并观察胸膜粘连情况,采用胸腔积液中纤维蛋白原(FBG)水平和胸腔粘连带数量评分评估大鼠胸腔积液粘连性;取胸膜组织标本,以体积分数10%甲醛溶液固定。测量各组大鼠切片中平面状态下的胸膜厚度,苏木精-伊红(HE)染色观察各组大鼠胸膜组织病理学改变,采用Western blot法检测各组大鼠胸膜组织中cGAS和STING蛋白表达,采用酶联免疫吸附试验法检测各组大鼠胸膜组织中cGAMP蛋白和胸腔积液中基质金属蛋白酶(MMP)-1、MMP-9、白细胞介素(IL)-1、IL-6、可溶性细胞间黏附分子-1(sICAM-1)水平。结果模型组和cGAS抑制剂组大鼠胸腔积液显著多于对照组,胸膜厚度显著大于对照组(P<0.05);cGAS抑制剂组大鼠胸腔积液显著少于模型组,胸膜厚度显著小于模型组(P<0.05)。cGAS抑制剂组大鼠胸腔积液中FBG水平、胸腔积液粘连性评分显著低于模型组(P<0.05)。HE染色显示,对照组大鼠肺间质和胸膜内血管排列、形态正常,胸膜无明�Objective To investigate the role of cyclic guanosine adenylate synthetase(cGAS)-cyclic guanosine monophosphate adenosine monophosphate(cGAMP)-stimulator of interferon gene(STING)signal pathway in rats with tuberculous pleurisy.Methods Thirty-six-week-old Sprague Dawley male rats without specific pathogen free were randomly divided into control group,model group and cGAS inhibitor group after adaptive feeding for 5 days,with 10 rats in each group.After 5 days of adaptive feeding,100μL(0.06 mg)bacillus calmette-guerin(BCG)suspension was injected intradermally into the right medial inguinal of rats in the three groups.Five weeks after inoculation,the rats in the model group and the cGAS inhibitor group were injected with 1 mL(0.03 mg)Mycobacterium tuberculosis H37RV suspension at the apex of the right angulus arcuum costarum to establish the models of tuberculous pleurisy,while the rats in the control group were only inoculated with BCG instead of Mycobacterium tuberculosis H37RV suspension.The rats in the cGAS inhibitor group were injected with 200μL cGAS inhibitor RU.521(600μg·L^(-1))by the vena caudalis from the second day of modeling,once a day for 7 days.The rats in the control group and model group were injected with the same volume of normal saline through the vena caudalis.On the eighth day,the rats were decapitated and killed,then the skin and sternum were immediately cut along the sternum from the side of the xiphoid process of the rats,and the thoracic cavity and mediastinum were exposed.The distribution of pleural effusion was observed,and the amount of pleural effusion was collected and recorded;the pleural adhesions were observed.The level of fibrinogen(FBG)in pleural effusion and the number of pleural adhesions were used to evaluate the adhesiveness of pleural effusion in rats.The distribution of pleural effusion of rats was observed,and the volume of pleural effusion was collected and recorded.The pleural adhesions were observed,and the adhesiveness of pleural effusion was evaluated by the level

关 键 词：结核性胸膜炎 环鸟苷酸腺苷酸 环鸟苷酸腺苷酸合成酶 干扰素基因刺激分子 

分 类 号：R521.7[医药卫生—内科学]