| 李峰,王罡艳,陶薪吉.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[J].安徽医药,2021,25(11):2205-2209. |
| 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 |
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| DOI：10.3969/j.issn.1009-6469.2021.11.019 |
| 中文关键词: 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 常组，)，模型，中， |
| 英文关键词: Pneumococcal Infections Suppressor of cytokine signaling 1 Streptococcus pneumoniae Alveolar epithelial cells Apoptosis |
| 基金项目: |
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| 中文摘要: |
| 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 pcDNA3.1SOCS1，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(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)释放率和细法胞中丙二醛(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(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)×103U/g，组(68.41±6.38)× 103U/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。 |
| 英文摘要: |
| Objective To study the effect of Suppressor of Cytokine Signal Transduction 1 (SOCS1) on the damage of alveolar A549epithelial 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 transfected into cells, stimulation of streptococcus pneumoniae was given, the expression of SOCS1 in cells was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR), the expression of SOCS1 protein was detected by Westernblotting, Methyl Thiazolyl Tetrazolium (MTT) was used to detect cell proliferation activity, lactate dehydrogenase (LDH) release rate andmalondialdehyde (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 byJC-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 activitydecreased [(100.00±9.68)% vs.(63.47±5.20)%], the LDH release rate increased [(2.68±0.27)% vs.(12.30±1.78)%], thecontent of MDA increased [(75.61±5.32) nmol/g vs. (165.92±13.17) nmol/g], SOD activity decreased [(131.47±12.86)×103U/g vs. (68.41±6.38)×103U/g], the apoptotic rate and the levels of C-caspase-3 and C-caspase-9 protein increased, the mitochondrial membrane potential decreased,Cytochrome Cprotein levelincy to plasmincreased,the level of Cytochrome Cproteininmi to chondriadecreased.Thee xpression of SO CS1ina lveolar A549 epithelial cells increased after pcDNA3.1-SOCS1 transfection, the cell activity increased, the LDH release rate decreased, thecontent of MDA decreased,SOD activity increased, the apoptotic rate and the levels ofC-caspase-3 and C-caspase-9 protein decreased,themitochondrial membrane potential increased, the level of Cytochrome C protein in mitochondria increased, Cytochrome C protein level incytoplasmdecreased.Conclusion SOCS 1alleviatesthedamageofalveolar A549epi the lialcellsinfectedby Streptococ cuspneu moniae. |
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