摘要
目的研究不同浓度苦参碱对常氧及低氧下人肺成纤维细胞株MRC-5的增殖及结缔组织生长因子(CTGF)、低氧诱导因子1α(HIF-1α)表达的影响。方法采用MRC-5常规体外培养传代,细胞培养至80%融合时,将细胞分组,分别在常氧(21%O2,74%N2,5%CO2)及低氧(1%O2,94%N2,5%CO2)下采用不同浓度苦参碱(终浓度0～3.2mmol/L)干预24h。按培养条件不同分为8组:常氧组(N0组)、常氧+苦参碱0.2mmol/L组(N0.2组)、常氧+苦参碱0.4mmol/L组(N0.4组)、常氧+苦参碱0.8mmol/L组(N0.8组)、低氧组(H0组)、低氧+苦参碱0.2mmol/L组(H0.2组)、低氧+苦参碱0.4mmol/L组(H0.4组)和低氧+苦参碱0.8mmol/L组(H0.8组)。采用四氮甲基唑蓝比色法(MTT法)检测细胞活性,采用Westernblot法检测细胞CTGF、HIF-1α蛋白表达情况。结果低氧对各组细胞增殖有促进作用(P<0.05);苦参碱对常氧及低氧下细胞增殖有抑制作用(P<0.05),具有浓度依赖性。CTGF、HIF-1α在常氧下有低水平表达,低氧下表达明显增强(P<0.01);苦参碱对常氧及低氧下CTGF、HIF-1α的表达均有抑制作用(P<0.05)。结论苦参碱对常氧及低氧下培养的MRC-5细胞增殖有抑制作用,对细胞CTGF和HIF-1α表达有抑制作用,且具有一定浓度依赖性。
Objective To investigate the effects of matrine on cell proliferation and expression of connective tissue growth factor（CTGF） and hypoxia inducible factor-1α（HIF-1α） of human lung fibroblast （WRC-5） in normoxia （21% 02, 74% N2, 5% CO2 ） and hypoxia （1% 02,94% N2, 5% CO2 ） conditions. Methods MRC-5 ceils were cultured and divided into differrent groups interfered with different dose of Matrine （final concentration of 0 -3.2 mmol/L） in normoxia or hypoxia for 24 h. Cells were divided into 8 groups according to culture conditions, ie. normoxia group （ No group） , normoxia ＋ matrine 0. 2 mmoL/L group（ No.2 group） , normoxia ＋ matrine 0. 4 mmoL/L group （ N0.4 group） , normoxia ＋ matrine 0. 8 mmol/L group（ Nos group） ,hypoxia group（ H0 group） ,hypoxia ＋ matrine 0. 2 mmol/L group（ H0.2 group） ,hypoxia ＋ matrine 0.4 mmol/L group（ H0.4 group） ,and hypoxia ＋ matrine 0. 8 mmoL/L group（ H0.8 group）. The MTT assay was used to measure the cell proliferation activity. Western-blot assay was used to examine the expression of CTGF and HIF-1α. Results Hypoxia promoted the cell proliferation in all groups （P 〈 0. 05）. Matrine inhibited the proliferation of WRC-5 cells in a concentration-dependent manner in hypoxia or normoxia conditions （P 〈 0. 05 ）. The expression of CTGF and HIF-1α was lower in normoxia and higher in hypoxia （P 〈 0. 01 ）. Matrine inhibited the expression of CTGF and HIF-1α in a concentration-dependent manner in hypoxia and normoxia（ P 〈 0. 05）. Conclusion Matrine can inhibit the cell proliferation and the expression of CTGF and HIF-1α of WRC-5 cells in normoxia and hypoxia in a concentration-dependent manner.
出处
《中国呼吸与危重监护杂志》
CAS
2010年第1期44-47,共4页
Chinese Journal of Respiratory and Critical Care Medicine
基金
四川省科技厅攻关课题(编号:0040205301469)
