CAJ | 학술논문

目的探讨辛伐他汀在体外对肺泡Ⅱ型细胞( ATⅡ)缺氧复氧损伤的保护作用及其机制.方法以人ATⅡ来源的A549细胞株为对象,应用化学性缺氧模拟剂CoCl2建立体外缺氧复氧损伤模型.研究分为空白组、对照组及不同剂量辛伐他汀治疗组(5、10、20、30、50、100μmol/L).应用CCK-8 比色法检测细胞增殖率；AV/PI流式细胞双染检测细胞凋亡率；Western blot法检测ATⅡ特异性标志表面蛋白C(SP-C)和增殖细胞核抗原(PCNA)的蛋白表达水平.结果与对照组相比,在A549细胞缺氧前应用低剂量辛伐他汀(5～20 μmol/L)预处理30 min,可促进A549细胞的增殖并显著抑制CoCl2引起的凋亡,同时显著增加SP-C和PCNA的蛋白表达；而高剂量辛伐他汀组(50～ 100 μmol/L)并没有观察到促进增殖、抑制凋亡的保护作用.给予L-甲羟戊酸竞争性抑制辛伐他汀,可逆转辛伐他汀对ATⅡ细胞的保护作用.结论低剂量辛伐他汀可逆转ATⅡ细胞缺氧复氧损伤,其保护机制与抑制甲羟戊酸代谢通路相关.
목적 탐토신벌타정재체외대폐포Ⅱ형세포( ATⅡ)결양복양손상적보호작용급기궤제.방법이인ATⅡ래원적A549세포주위대상,응용화학성결양모의제CoCl2건입체외결양복양손상모형.연구분위공백조、대조조급불동제량신벌타정치료조(5、10、20、30、50、100μmol/L).응용CCK-8 비색법검측세포증식솔；AV/PI류식세포쌍염검측세포조망솔；Western blot법검측ATⅡ특이성표지표면단백C(SP-C)화증식세포핵항원(PCNA)적단백표체수평.결과 여대조조상비,재A549세포결양전응용저제량신벌타정(5～20 μmol/L)예처리30 min,가촉진A549세포적증식병현저억제CoCl2인기적조망,동시현저증가SP-C화PCNA적단백표체；이고제량신벌타정조(50～ 100 μmol/L)병몰유관찰도촉진증식、억제조망적보호작용.급여L-갑간무산경쟁성억제신벌타정,가역전신벌타정대ATⅡ세포적보호작용.결론 저제량신벌타정가역전ATⅡ세포결양복양손상,기보호궤제여억제갑간무산대사통로상관.
Objective To investigate the protective effects of simvastatin on cobalt choride ( CoCl2 ) -induced hypoxia and reoxygenation injury on alveolar type Ⅱ cells and the underlying mechanisms.Methods CoCl2 was used to establish the hypoxia and reoxygenation injury model on AT Ⅱ cells.Blank,control and variant doses simvastatin-treated groups ( 5,10,20,30,50,100 μ mol/L) were designed in the present study.The proliferation of AT Ⅱ cells was evaluated by Cell Counting Kit-8 ( CCK-8 ) assay.The percentage of apoptotic cells was assessed by flow cytometry AV/PI double-staining.The protein levels of surfactant protein-C (SP-C) and proliferating cell nuclear antigen (PCNA) in AT Ⅱ cells was determined by Western blot.Results As compared with the control group,pretreatment with low dose (5 - 20 μmol/L),but not high dose simvastatin (50 - 100 μmol/L) markedly reduced A549 cells apoptosis,and increased their proliferation and the protein levels of SPC and PCNAin vitro.The protective effect could be reversed in vitro by L-mevalonate,a simvastatin competitive inhibitor,which indicated that the inhibition of mevalorate pathway was involved in the simvastatin induced AT Ⅱ cells function restoration.Condusion Low doses simvastatin reversed CoCl2-induced hypoxia and reoxygenation injury of AT Ⅱ cells.The inhibition of mevalonate pathway contributed to simvastatin induced AT Ⅱ cells function restoration.