棕榈酸诱导人绒毛滋养层细胞胰岛素抵抗模型的建立

摘要

目的 探讨人胎盘绒毛滋养层细胞HTR8-S/Vneo在高脂环境下是否会形成胰岛素抵抗. 方法 用MTT法测定棕榈酸和胰岛素不影响HTR8-S/Vneo细胞增殖的最适浓度,在该最适浓度下分为四组(对照组、棕榈酸组、胰岛素组、棕榈酸-胰岛素协同处理组)处理HTR8-S/Vneo细胞,检测不同处理条件及不同处理时间下,细胞葡萄糖摄取量和细胞内总糖量. 结果 用MTT法检测不同浓度棕榈酸及胰岛素处理的HTR8-S/Vneo细胞,发现棕榈酸浓度为0.125 mmol/L、胰岛素浓度为100 nmol/L时,对细胞增殖没有显著影响(P＞0.05),故选择该浓度的棕榈酸与胰岛素处理HTR8-S/Vneo细胞.在处理12h、18h、24 h、36h分别检测葡萄糖的消耗量和细胞内总糖量,发现与对照组相比,胰岛素处理后葡萄糖消耗量和细胞内总糖量呈上升趋势(P＜0.05),而棕榈酸处理后葡萄糖消耗量和细胞内总糖量无显著变化(P＞0.05);与棕榈酸单独处理组相比,棕榈酸-胰岛素协同处理组在12h和18h能显著增加细胞内总糖量(P＜0.05),但在24 h和36 h时这种显著差异消失(P＞0.05),且与单独胰岛素处理组相比,棕榈酸-胰岛素协同处理组显著降低了葡萄糖消耗量和细胞内总糖量(P＜0.05).结论 棕榈酸-胰岛素协同处理24 h,引起HTR8-S/Vneo细胞对胰岛素的敏感性降低,HTR8-S/Vneo细胞胰岛素抵抗模型建立;与文献报道模型相比,棕榈酸-胰岛素协同处理大大缩短了细胞出现胰岛素抵抗的时间.%Objective:To investigate whether the high palmitic acid(PA)can induce the placental trophoblast cells(HTR8-S/Vneo)to form insulin resistance in high fat environment.Methods:The optimum concentrations of palmitic acid and insulin which do not affect the proliferation of HTR8-S/Vneo cells were tested by MTT assay.According to the optimum concentrations obtained by the MTT assay,the HTR8-S/Vneo cells were divided into four groups(control group,PA group,insulin group,PA-insulin co-treatment group).The cell glucose uptake and intracellular total sugar were determined in the different treatment conditions and different processing time.Results:The MTT assay in the HTR8-S/Vneo cells treated with different concentrations of PA and insulin found that PA concentration of 0.125 mmol/L and insulin concentration of 100 nmol/L had no significant effect on cell proliferation(P＞0.05),so the concentration of PA and insulin was chosen to treat HTR8-S/Vneo cells.Glucose consumption and total intracellular total sugar were measured at 12,18,24 and 36 hours after treatment.The experiment results found glucose consumption and intracellular total sugar were increased after insulin treatment compared with the control group(P＜0.05),but there was no significant changes in glucose consumption and intracellular total sugar content after PA treatment(P＜0.05).Compared with PA treatment alone group,PA-insulin synergistic treatment group could significantly increase cells at 12 and 18 hours(P＜0.05),but the significant difference was disappeared at 24 and 36 hours(P＞0.05).Compared with insulin alone group,glucose consumption and total cellular sugar were significantly reduced in PA-insulin co-treatment group(P＜0.05).Conclusions:The synergistic effect of palmitic acid and insulin for 24 hours results in the decrease of insulin sensitivity in HTR8-S/Vneo cells to establish the insulin resistance model in HTR8-S/Vneo cells.Compared with the reported model,palmitic acid-insulin synergistic treatment can greatly shorten the time of insulin resistance in cells.