金丝桃苷下调微小RNA-199a对脂多糖诱导的人肺泡上皮细胞的影响  

Effect of Hypericin on LPS-induced HPAEpiC cells by down-regulating miR-199a

作　　者：申玲君[1] 张海瑞[1] 邵伟[1] 冯振霞 邬超[2] SHEN Lingjun;ZHANG Hairui;SHAO Wei;FENG Zhenxia;WU Chao(Department of Respiratory and Critical Care Medicine,Anyang District Hospital,Puyang City,Anyang,Henan 455000 China;Department of Respiratory,People's Hospital of Xinjiang Uygur Autonomous Region,Urumqi,Xinjiang Uygur Autonomous Region 830002,China)

机构地区：[1]濮阳市安阳地区医院呼吸与危重症医学科,河南安阳455000 [2]新疆维吾尔自治区人民医院呼吸内科,新疆维吾尔自治区乌鲁木齐830002

出　　处：《安徽医药》2022年第2期217-221,共5页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨金丝桃苷对脂多糖(LPS)诱导的人肺泡上皮细胞(HPAEpiC)的影响及分子机制。方法将HPAEpiC分为对照组、模型组、金丝桃苷低、中、高剂量药物组、anti-miR-NC组、anti-miR-199a组、高剂量药物组+miR-NC组、高剂量药物组+miR-199a组。四甲基偶氮唑盐比色法(MTT)检测细胞增殖;流式细胞术检测细胞周期和细胞凋亡情况;酶联免疫吸附试验(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平;实时荧光定量PCR(RT-qPCR)检测miR-199a表达水平;蛋白质印迹(Western blotting)法检测裂解的半胱天冬蛋白酶3(Cleaved-caspase3)、半胱天冬蛋白酶3前体(pro-caspase3)的蛋白表达。结果金丝桃苷处理后,LPS诱导的HPAEpiC中细胞OD值升高,金丝桃苷低、中、高剂量药物组OD值分别为(0.49±0.02)、(0.60±0.03)、(0.72±0.03),G0/G1期细胞所占比例降低,S期细胞所占比例升高,细胞凋亡率降低,TNF-α、IL-1β水平降低,miR-199a表达水平降低,Cleaved-caspase3表达水平降低,pro-caspase3表达水平升高,且呈剂量依赖性(P<0.05);抑制miR-199a表达可抑制LPS诱导的HPAEpiC细胞凋亡和炎症因子的释放。且miR-199a过表达逆转了金丝桃苷提取物对LPS诱导的HPAEpiC细胞凋亡和炎症因子的作用。结论金丝桃苷可能通过下调miR-199a抑制LPS诱导的HPAEpiC细胞凋亡和炎症反应。Objective To investigate the effect and molecular mechanism of hyperoside on HPAEpiC cells induced by LPS.Meth⁃ods Human alveolar epithelial cells(HPAEpiC)were divided into control group,model group,hypericin low,medium and high dose drug group,anti-miR-NC group,anti-miR-199a group,high dose drug group+miR-NC group,high-dose drug group+miR-199a group.Tetramethylazoazole colorimetric method(MTT)was used to detect cell proliferation;flow cytometry was used to detect cell cycle and apoptosis;enzyme-linked immunosorbent assay(ELISA)was used to detect tumor necrosis factor-α(TNF-α),Interleukin-1β(IL-1β)lev⁃el;real-time fluorescent quantitative PCR(RT-qPCR)to detect miR-199a expression;Western blotting method to detect protein expres⁃sion of cleaved cysteine-containing aspartate-specific proteases 3(Cleaved-caspase3),caspase 3 precursor(pro-caspase3).Results Af⁃ter treatment with low,medium and high doses of hyperoside,the cells OD value in HPAEpiC induced by LPS was increased[(0.49±0.02),(0.60±0.03),(0.72±0.03)],the proportion of cells in G0-G1 phase was decreased,the proportion of cells in S phase was increased,the apoptosis rate was decreased,and the levels of TNF-αand IL-1βwere decreased,the expression of miR-199a was decreased,the expression of Cleaned-caspase3 was decreased,and the expression of pro-caspase3 was increased,in a dose-dependent manner(P<0.05).Inhibition of miR-199a expression can inhibit LPS-induced apoptosis and release of inflammatory factors in HPAEpiC cells.Over⁃expression of miR-199a reversed the effect of hyperoside extract on LPS-induced apoptosis and inflammatory factors in HPAEpiC cells.Conclusion Hypericin may inhibit LPS-induced apoptosis and inflammation of HPAEpiC cells by down-regulating miR-199a.

关 键 词：金丝桃属 金丝桃苷 微小RNA-199a 肺泡上皮细胞 急性肺损伤 炎症 

分 类 号：R285[医药卫生—中药学]