玄参提取物调控circ_0038467/miR-495对肺炎链球菌诱导的肺泡上皮细胞损伤的影响  

Effect of extract from scrophulariae on damage of alveolar epithelial cells induced by Streptococcus pneumoniae regulating by circ_0038467/miR-495

作　　者：余莉萍[1] 余淑菁 李旭成[1] 崔金涛[1] 陆佳鑫 YU Liping;YU Shujing;LI Xucheng;CUI Jintao;LU Jiaxin(Department of Emergency,Wuhan Hospital of Traditional Chinese Medicine,Wuhan 430000,China)

机构地区：[1]武汉市中医医院急诊科,武汉430000 [2]武汉市优抚医院,武汉430000

出　　处：《中国免疫学杂志》2023年第2期257-260,266,共5页Chinese Journal of Immunology

基　　金：国家中医药管理局资助项目(国中医药人教函[2019]41号);湖北省卫生健康委员会资助项目(ZY2019M048)。

摘　　要：目的:探讨玄参提取物对肺炎链球菌诱导的肺泡上皮细胞损伤的影响及作用机制。方法:采用肺炎链球菌诱导肺泡上皮细胞建立细胞损伤模型并给予不同剂量玄参提取物处理。si-NC、si-circ_0038467分别转染肺泡上皮细胞,1×108 CFU/ml肺炎链球菌处理,pcDNA-circ_0038467转染肺泡上皮细胞,40µg/ml玄参提取物与1×108 CFU/ml肺炎链球菌处理;MTT、流式细胞术分别检测细胞增殖及凋亡;ELISA检测IL-6、IL-10水平;qRT-PCR检测circ_0038467、miR-495表达;双荧光素酶报告实验检测circ_0038467与miR-495的靶向关系;Western blot检测Cleaved-caspase3蛋白表达。结果:玄参提取物可浓度依赖性地增强肺炎链球菌诱导的肺泡上皮细胞活力(P<0.05),并提高miR-495表达和IL-10水平(P<0.05),而降低circ_0038467表达、IL-6水平、细胞凋亡率和Cleaved-caspase3蛋白表达(P<0.05);转染si-circ_0038467可促进miR-495表达(P<0.05),提高细胞活力和IL-10水平(P<0.05),降低IL-6水平、细胞凋亡率和Cleaved-caspase3蛋白水平(P<0.05);circ_0038467可靶向调控miR-495表达及活性;转染pcDNA-circ_0038467可减弱玄参提取物对细胞增殖、凋亡及炎症的作用。结论:玄参提取物可通过调控circ_0038467/miR-495表达促进细胞增殖,抑制细胞凋亡及炎症反应,从而减轻肺炎链球菌诱导的肺泡上皮细胞损伤。Objective:To explore effect of extract from scrophulariae on alveolar epithelial cell injury induced by Streptococcus pneumoniae and its mechanism.Methods:Streptococcus pneumoniae was used to induce alveolar epithelial cells to establish a cell injury model and treated with different doses of extract from scrophulariae.si-NC and si-circ_0038467 were respectively transfected into alveolar epithelial cells and treated with 1×108 CFU/ml Streptococcus pneumoniae.pcDNA-circ_0038467 was transfected into alveolar epithelial cells and treated with 40μg/ml extract from scrophulariae and 1×108 CFU/ml Streptococcus pneumoniae.MTT and flow cytometry were used to detect cell proliferation and apoptosis.ELISA was used to detect levels of IL-6 and IL-10.qRT-PCR was used to detect expressions of circ_0038467 and miR-495.Dual luciferase reporter experiment was used to detect targeting relationship between circ_0038467 and miR-495.Western blot was used to detect expression of Cleaved-caspase3 protein.Results:Extract from scrophulariae could enhance viability of alveolar epithelial cells induced by Streptococcus pneumoniae in a concentration-dependent manner(P<0.05),and increase expression of miR-495 and level of IL-10(P<0.05),while reduce expression of circ_0038467,level of IL-6,cell apoptosis rate and protein level of Cleaved-caspase3(P<0.05).Transfection of si-circ_0038467 could promote expression of miR-495(P<0.05),increase cell viability and level of IL-10(P<0.05),but decrease level of IL-6,cell apoptosis rate and protein level of Cleaved-caspase3(P<0.05).circ_0038467 could target expression and activity of miR-495.Transfection of pcDNA-circ_0038467 could attenuate effects of extract from scrophulariae on cell proliferation,apoptosis and inflammation.Conclusion:Extract from scrophulariae can promote cell proliferation,inhibit cell apoptosis and inflammation by regulating expression of circ_0038467/miR-495,thereby reducing damage of alveolar epithelial cells induced by Streptococcus pneumoniae.

关 键 词：玄参提取物 circ_0038467 miR-495 肺炎链球菌 肺泡上皮细胞 细胞增殖 凋亡 

分 类 号：R378.1[医药卫生—病原生物学]