不同溶剂中油酸对LO_2和HepG_2细胞生长和活力的影响

Influence of oleic acid in different solvent media on LO_2 and HepG_2 cell-growth and-viability

目的探讨以二甲基亚砜(DMSO)和牛血清白蛋白(BSA)分别溶解的油酸(OA)对人正常肝细胞(LO2)株和人肝癌细胞(HepG2)株的影响,寻找合适的脂肪酸溶剂以及体外研究脂代谢的肝细胞系。方法 LO2和HepG2细胞均分为:空白组,DMSO组,DMSO+不同浓度油酸组,BSA组,BSA+不同浓度油酸组。培养48 h后,采用MTT法检测细胞活力,油红O染色观察活细胞数目和胞内脂滴变化;甘油-3-磷酸氧化酶法检测细胞内甘油三酯(TG)的含量。结果以DMSO溶解的(0.4～6.4)μg/ml油酸分别处理的LO2和HepG2细胞,细胞生长曲线均呈先上升后下降的趋势,除1.6μg/ml浓度组LO2细胞生长接近空白对照组外,其它组细胞活力均显著低于对照组(P<0.05);同样以BSA溶解的(0.4～6.4)μg/ml油酸分别处理LO2和HepG2细胞,各组细胞活力均高于空白组,呈上升趋势(P<0.01)。油红O染色显示两细胞株在DMSO溶解OA处理后,与空白组比较,油酸组细胞内脂滴增多,但细胞数目减少;而在BSA溶解下,细胞内脂滴和活细胞数目均随OA浓度增加而增加,各组细胞内TG含量变化趋势与油红O染色结果类似,以LO2细胞株更为敏感。结论作为OA溶剂,BSA较DMSO合适;LO2较HepG2细胞更适合作为体外研究脂代谢的细胞株。

Objective To find a suitable media for fatty acids and a convenient cell line in vitro for studying lipid metabolism, by comparing the effect of oleic acid （OA） at different solvent media on human liver cells LO2 and hepato-carcinoma ceils HepG2. Methods Both LO2 and HepG2 cell lines were divided into normal group, DMSO group, groups of different concentrations of oleic acid, BSA group, and groups of different concentrations of oleic acid. After being cultured for 48 hours, the cell viability was analyzed with MTI＇ method, the fatty droplets in cells could be observed by oil red O staining under optical microscope. The intracellular triglyceride （TG） was detected by reactant kits. Results The growth curves of the two cell lines, which were treated with different concentrations of oleic acid （0. 4, 0. 8, 1.6, 3.2, 6.4μg/ml） dissolved in DMSO, showing first increased and then a decreased trend. Except the group of LO2 with 1.6 μg / ml OA, the cell viability of the other groups was significantly lower than that of the normal group （ P 〈 0.05 ）. However, the viability of the two cell lines, treated with oleic acid （0.4, 0. 8, 1.6, 3.2, 6.4μg/ml） dissolved in BSA, were higher than that of the normal group, and the curves showed an upward trend. Oil red O staining showed that the intracellular lipid droplets of the two cell lines treated with oleic acid dissolved in DMSO increased, as compared with the normal group, and the cell number decreased, but the lipid droplets and the cell number of the two cell lines treated with oleic acid dissolved in BSA both increased. The level of TG in each group was similar to the trend of the oil red O staining, while LO2 cell line was more sensitive. Conclusion Compared with DMSO, BSA is a better dissolution media for fat acids, and LO2 cell line was more suitable for the development in vitro model of liver lipid metabolism .