代综方对3T3-L1脂肪细胞胰岛素抵抗模型瘦素、抵抗素及肿瘤坏死因子α的影响

Effect of Dai-Zong-Fang on Leptin,Resistin,and TNF-α in 3T3-L1 Adipocyte Insulin Resistance Model

目的观察中药代综方对棕榈酸(Palmitic acid,PA)诱导胰岛素抵抗(Insulin resistance,IR)的3T3-L1脂肪细胞中细胞因子瘦素、抵抗素、肿瘤坏死因子α(Tumor necrosis factor-α,TNF-α)的影响。方法采用传统“鸡尾酒式”诱导3T3-L1前脂肪细胞分化成熟,用0.5 mmol/L PA建立IR模型。分为空白对照组、模型组、代综方低浓度组(含代综方生药量0.5 mg/ml)、代综方高浓度组(含代综方生药量2.0 mg/ml),干预48 h。葡萄糖氧化酶法检测培养上清中的葡萄糖浓度,计算葡萄糖基础消耗量(G_(Basic))和胰岛素刺激下消耗量(G_(Ins)),评价胰岛素敏感性指数(ISI)。ELISA检测上清中瘦素、抵抗素、TNF-α的含量,Real-Time PCR检测瘦素、抵抗素、TNF-α的mRNA表达。结果与空白对照组比较,模型组GIns、ISI均明显降低(P<0.01);与模型组比较,代综方高、低浓度组G_(Basic)、G_(Ins)、ISI均明显升高(P<0.05,P<0.01),且代综方高浓度组G_(Basic)、G_(Ins)、ISI较代综方低浓度组明显升高(P<0.05,P<0.01);与模型组比较,代综方高浓度组3T3-L1脂肪细胞上清中瘦素的含量明显增加(P<0.01),且抵抗素、TNF-α的含量明显减少(P<0.05,P<0.01);与模型组比较,代综方高浓度组3T3-L1脂肪细胞瘦素mRNA表达水平明显升高(P<0.01),且代综方低浓度和高浓度组抵抗素、TNF-αmRNA表达水平均明显降低(P<0.05,P<0.01)。结论代综方能够增加PA诱导的IR脂肪细胞葡萄糖消耗,提高胰岛素敏感性,同时上调瘦素水平,降低抵抗素、TNF-α水平。

Objective To observe the effect of Dai-Zong-Fang(DZF)on leptin,resistin,and tumor necrosis factorα(TNF-α)in the 3 T3-L1 adipocyte of insulin resistance(IR)induced by palmitic acid(PA).Methods 3 T3-L1 preadipocytes were induced to differentiate and mature by traditional“cocktail”method,and the IR model was established by 0.5 mmol/L PA.The experiment was divided into a blank control group,a model group,a DZF low-concentration group(0.5 mg/ml),and a DZF high-concentration group(2.0 mg/ml).The intervention lasted for 48 h.Glucose concentration in the culture supernatant was measured by glucose oxidase method.The basic glucose consumption(G)and the insulin-stimulated glucose consumption(G)were calculated to evaluate insulin sensitivity index(ISI).The levels of leptin,resistin,and TNF-αin the supernatant were detected by enzyme-linked immunosorbent assay(ELISA),and the corresponding mRNA expressions were detected by real-time quantitative polymerase chain reaction(real-time PCR).Results The Gand ISI in the model group were significantly lower as compared with the blank control group(P<0.01).The G_(Basic),G_(Ins),and ISI in the DZF high and low-concentration groups were higher as compared with the model group(P<0.05,P<0.01),and the G_(Basic),G_(Ins),and ISI in the DZF high-concentration group were higher than those in the DZF low-concentration group(P<0.05,P<0.01).As compared with the model group,the content of leptin in the 3 T3-L1 adipocyte supernatant of the DZF high-concentration group increased(P<0.01),whereas the content of resistin and TNF-αreduced(P<0.05,P<0.01).As compared with the model group,the mRNA expression level of leptin in the 3 T3-L1 adipocytes in the DZF high-concentration group increased(P<0.01),and the mRNA expression levels of resistin and TNF-αin the DZF low and high groups decreased(P<0.05,P<0.01).Conclusion DZF increases the glucoseconsumption of IR adipocytes induced by PA,and improves the insulin sensitivity.At the same time,it up-regulates the level of leptin,and down-regulates the levels ofresistin and TNF-α.