| 申玲君,张海瑞,邵伟,等.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[J].安徽医药,2022,26(2):217-221. |
| 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 |
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| DOI：10.3969/j.issn.1009-6469.2022.02.002 |
| 中文关键词: 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 |
| 英文关键词: Hypericum Hyperoside MiR-199a Alveolar epithelial cells Acute lung injury Inflammation |
| 基金项目:国家自然科学基金资助项目( 81960005) |
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| 中文摘要: |
| 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；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)的蛋白表蛋达。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)、 |
| 英文摘要: |
| 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 dosedrug 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 andapoptosis; 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-α and IL-1β were 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. |
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