细胞因子信号转导抑制因子1对肺炎链球菌感染的肺泡上皮细胞A549损伤的影响  

Effect of suppressor of cytokine signal transduction 1 on the damage of alveolar epithelial cells A549 infected by streptococcus pneumoniae

作　　者：李峰 王罡艳 陶薪吉 LI Feng;WANG Gangyan;TAO Xinji(Author Affiliation:Department of Critical Care Medicine,Wuhan Eighth Hospital,Wuhan,Hubei 430010,China)

机构地区：[1]武汉市第八医院重症医学科,湖北武汉430010

出　　处：《安徽医药》2021年第11期2205-2209,共5页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨细胞因子信号转导抑制因子1(SOCS1)对肺炎链球菌感染的肺泡上皮细胞A549损伤的影响。方法以肺泡上皮细胞A549为研究对象,A549细胞分成正常组、模型组、Vector+模型组、SOCS1+模型组共四组,在细胞中转染pcDNA3.1-SOCS1,给予肺炎链球菌刺激,实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测细胞中SOCS1 mRNA表达水平,蛋白质印迹法(Western blotting)检测SOCS1蛋白表达水平,噻唑蓝(MTT)检测细胞增殖活性,试剂盒检测细胞乳酸脱氢酶(LDH)释放率和细胞中丙二醛(MDA)、超氧化物歧化酶(SOD)水平,流式细胞术检测细胞凋亡,Western blotting检测活化的含半胱氨酸的天冬氨酸蛋白水解酶-3(C-caspase-3)、活化的含半胱氨酸的天冬氨酸蛋白水解酶-9(C-caspase-9)蛋白水平,JC-1法检测线粒体膜电位,Western blotting检测胞质和线粒体中细胞色素C(Cytochrome C)蛋白水平。结果肺炎链球菌刺激后的肺泡上皮细胞A549中SOCS1表达水平下降[正常组SOCS1 mRNA(1.00±0.09),模型组SOCS1 mRNA(0.59±0.07)],细胞存活率降低[正常组(100.00±9.68)%,模型组(63.47±5.20)%],LDH释放率升高[正常组(2.68±0.27)%,模型组(12.30±1.78)%],MDA含量升高[正常组(75.61±5.32)nmol/g,模型组(165.92±13.17)nmol/g],SOD活性降低[正常组(131.47±12.86)×103 U/g,模型组(68.41±6.38)×103 U/g],细胞凋亡率和细胞中C-caspase-3、C-caspase-9蛋白水平升高,线粒体膜电位下降,胞质中Cytochrome C蛋白水平升高,线粒体中Cytochrome C蛋白水平下降。pcDNA3.1-SOCS1转染后的肺炎链球菌条件下肺泡上皮细胞A549中SOCS1表达水平升高,细胞活性升高,LDH释放率降低,MDA含量下降,SOD活性升高,细胞凋亡率和细胞中C-caspase-3、C-caspase-9蛋白水平降低,线粒体膜电位升高,线粒体中Cytochrome C蛋白水平升高,胞质中Cytochrome C蛋白水平降低。结论SOCS1减轻肺炎链球菌感染的肺泡上皮细胞A549损伤。Objective To study the effect of Suppressor of Cytokine Signal Transduction 1(SOCS1)on the damage of alveolar A549 epithelial cells infected by Streptococcus pneumoniae.Methods The alveolar A549 epithelial cells were selected as the research object,and assigned into four groups:normal group,model group,Vector+model group,and SOCS1+model group.pcDNA3.1-SOCS1 was trans‐fected into cells,stimulation of streptococcus pneumoniae was given,the expression of SOCS1 in cells was detected by real-time fluores‐cence quantitative reverse transcription polymerase chain reaction(qRT-PCR),the expression of SOCS1 protein was detected by Western blotting,Methyl Thiazolyl Tetrazolium(MTT)was used to detect cell proliferation activity,lactate dehydrogenase(LDH)release rate and malondialdehyde(MDA)and superoxide dismutase(SOD)levels were detected by the kit,cell apoptosis was detected by flow cytometry,Western blotting was used to detect C-caspase-3 and C-caspase-9 protein levels,the mitochondrial membrane potential was measured by JC-1 method,Cytochrome C protein levels in cytoplasm and mitochondria were detected by Western blotting.Results The expression of SOCS1 in alveolar A549 epithelial cells stimulated by Streptococcus pneumoniae decreased[(1.00±0.09)vs.(0.59±0.07)],the cell activity decreased[(100.00±9.68)%vs.(63.47±5.20)%],the LDH release rate increased[(2.68±0.27)%vs.(12.30±1.78)%],the content of MDA in‐creased[(75.61±5.32)nmol/g vs.(165.92±13.17)nmol/g],SOD activity decreased[(131.47±12.86)×103 U/g vs.(68.41±6.38)×103 U/g],the apoptotic rate and the levels of C-caspase-3 and C-caspase-9 protein increased,the mitochondrial membrane potential decreased,Cyto‐chrome C protein level in cytoplasm increased,the level of Cytochrome C protein in mitochondria decreased.The expression of SOCS1 in al‐veolar A549 epithelial cells increased after pcDNA3.1-SOCS1 transfection,the cell activity increased,the LDH release rate decreased,the content of MDA decreased,SOD activity increased,the apoptotic rate and the leve

关 键 词：肺炎球菌感染 细胞因子信号转导抑制因子1 肺炎链球菌 肺泡上皮细胞 凋亡 

分 类 号：R563.1[医药卫生—呼吸系统]