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氧化型低密度脂蛋白对人主动脉平滑肌细胞生长的影响
康永波,蔡悦,李昱,杜余辉
0
(山西医科大学基础医学院;昆明理工大学医学院)
摘要:
【摘要】目的 探讨氧化型低密度脂蛋白(ox-LDL)作用不同时间对人主动脉平滑肌细胞(HA-VSMC)生长的影响,分析其诱导HAVSMC形成泡沫细胞的最适诱导浓度和时间,为开展动脉粥样硬化(AS)及相关疾病研究建立体外细胞模型提供依据。方法 使用不同浓度的oxLDL(15 μg/mL和20 μg/mL)分别处理HAVSMC不同时间(36、48、60、72、84、96 h),通过测定细胞内总胆固醇和胆固醇酯含量以及使用油红O染色观察细胞内脂质空泡来检测泡沫细胞形成的情况;用噻唑蓝比色实验(MTT法)检测细胞活力变化的情况。结果 细胞内总胆固醇和胆固醇酯含量、胆固醇酯/总胆固醇的比值以及脂质空泡随着oxLDL浓度和处理时间的增加而增加。15 μg/mL oxLDL作用60 h细胞内胆固醇酯/总胆固醇的比值超过50%且脂质空泡形成,表明泡沫细胞形成;同时,与对照组相比,15 μg/mL oxLDL作用36、48 h对细胞增殖有促进作用,细胞活力显著增加(P<005);处理60、72和84 h细胞活力未发生显著性变化;处理96 h对细胞增殖有抑制作用,细胞活力显著下降(P<005)。表明15 μg/mL oxLDL处理细胞建模不成功。20 μg/mL oxLDL作用48 h细胞内胆固醇酯/总胆固醇的比值超过50%且脂质空泡形成,表明泡沫细胞形成;同时,与15 μg/mL oxLDL处理的HAVSMC相比,20 μg/mL oxLDL处理细胞36 h细胞活力显著降低(P<001),处理48、60、72和84h细胞活力显著增加(P<005),处理96 h细胞活力无显著变化。20 μg/mL oxLDL处理细胞48、60、72和84 h建模成功。结论 浓度为20 μg/mL的 oxLDL处理HAVSMC细胞48 h细胞活力增加最为显著,其可作为建立AS体外细胞模型的最佳诱导时间和浓度。
关键词:  氧化型低密度脂蛋白  人主动脉平滑肌细胞  总胆固醇  胆固醇酯  细胞活力
DOI:
基金项目:山西医科大学省级博士启动基金项目(SD1807);山西医科大学校级博士启动基金项目
The Influence of oxidized low density lipoprotein on growth of human arterial smooth muscle cell in vitro
KANG Yongbo,CAI Yue,LI Yu,DU Yuhui
(School of Basic Medical Sciences, Shanxi Medical University;Medical Faculty, Kunming University of Science and Technology)
Abstract:
【Abstract】Objective To investigate the effects of different concentrations of oxidized lowdensity lipoprotein (oxLDL) on the growth of human aortic smooth muscle cells (HAVSMC) at different time in vitro, analyze the optimal concentration and time of oxLDL to induce HAVSMC forming foam cells, and provide a basis for establishing an in vitro cell models for atherosclerosis and related disease research. Methods In this study, different concentrations of oxLDL (15 μg/ml and 20 μg/ml) were used to treat VSMC at different times (36 h, 48 h, 60 h, 72 h, 84 h and 96 h). The formation of foam cells were evaluated by determining the total cholesterol and cholesterol ester content as well as by oil red O staining to observe the intracellular lipid vacuoles. Meanwhile, the change of cell viability was detected by thiazolyl blue colorimetric assay (MTT assay). Results The content of total cholesterol and cholesterol ester, the ratio of cholesterol ester to total cholesterol, and lipid vacuoles increased with the increase of ox LDL concentration and treatment time. The ratio of cholesteryl ester / total cholesterol in 15 h g/mL ox LDL was more than 50% and the formation of lipid vacuoles indicated foam cell formation. At the same time, compared with the control group, the effect of 15 g/mL ox LDL on 36 and 48 h promoted cell proliferation, and the cell viability was significantly increased (P<005). The vitality of 60, 72 and 84 h cells did not change significantly. The treatment of H H pairs showed that there was no significant change in the cell viability of the 60 h cells. Cell proliferation was inhibited and cell viability decreased significantly (P<005), which indicated that the cell model of 15 μg/mL ox LDL treatment was not successful. The ratio of cholesteryl ester/total cholesterol in 20 h g/mL ox LDL was more than 50% and the formation of lipid vacuoles indicated the formation of foam cells. At the same time, compared with HA VSMC treated with 15 g/mL ox LDL, the activity of 36 36 cells in 20 g/mL g/mL ox treatment cells was significantly reduced (0, 01), 48, 60, 72 and LDL cells increased significantly. There was no significant change in 96 h cell viability. The cells were treated with 20 μg/mL ox LDL for 48, 60, 72 and 84 hours.Conclusion The activity of HA VSMC cells treated with ox LDL at a concentration of 20 μg/mL for 48 hours increased most significantly, which could be used as the best induction time and concentration for establishing as cell model in vitro.
Key words:  Oxidized low density lipoprotein  Human arterial smooth muscle cell  Total cholesterol  Cholesterol ester  Cell viability

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