摘要
目的采用肿瘤坏死因子-α(TNF-α)体外诱导培养法建立大鼠骨骼肌细胞胰岛素抵抗(IR)模型,为研究IR的发生机制及筛选改善IR的药物和功能食品提供一种简便可靠的IR细胞模型。方法以大鼠肌细胞系L6细胞为研究对象,在含10ng/ml小鼠TNF-α培养液中培养24h,同时设立空白对照组和胰岛素对照组。之后各组细胞分别加入100nmol/L胰岛素。采用葡萄糖氧化酶-过氧化物酶法(GOD-POD)法检测培养液中残存葡萄糖含量,[3H]标记的脱氧葡萄糖转运检测细胞内葡萄糖的摄取量评价模型建立的效果。结果在无胰岛素刺激下,TNFα诱导组培养液中残存葡萄糖含量和细胞内的葡萄糖摄取量,与空白对照组比较没有差异(P>0.05);在胰岛素刺激下,TNFα诱导组培养液中残存葡萄糖含量和细胞内的葡萄糖摄取量,与胰岛素对照组比较差异具有显著性(P<0.05)。结论用含10ng/ml TNFα的培养液培养24h的L6细胞产生胰岛素抵抗,作为IR肌细胞模型可广泛用于胰岛素抵抗的机制研究和筛选辅助降血糖的药物或功能食品的功能评价。
Objective To establish a convenient and reliable insulin - resistant cell model induced by TNF-α for studying the mechanisms of insulin-resistance and screening medical drugs or functional foods assisted in improving insulin-resistance. Methods L6 cells in TNF-α group,insulin group and TNF-α plus insulin group wereincubated in culture medium containing 10ng /ml TNF-α,100 nmol /L insulin and both 10ng /ml TNF-α and 100 nmol /L insulin respectively for 24 hours. The glucose remained in culture medium was measured by the method of glucose oxidizes peroxides （GOD-POD）. The glucose uptake of L6 cells was measured by [3 H]-2-deoxyglucose. Results With no insulin stimulation,the glucose remained in culture medium in TNF-αgroup was equal to that in the control group （P 0. 05. ）. With insulin stimulation,the glucose remained in culture medium in insulin group was much lower than that in TNF-α group （ P 0. 05）. Conclusion The insulin resistance in L6 cells could be induced by high concentration of TNF-α. The IR model of skeletal muscle cell could be widely used for studying the mechanism of insulin resistance and for screening medical drugs and functional food assisted in reducing blood sugar.
出处
《卫生研究》
CAS
CSCD
北大核心
2010年第2期149-151,共3页
Journal of Hygiene Research
基金
国家"十一五"科技支撑计划项目(No.2006BAD28B01)
