山茱萸提取物对3T3-L1脂肪细胞葡萄糖消耗与脂肪分解的影响

Corni Fructus Extract Increased Glucose Consumption and Inhibited Isoproterenol-induced Lipolysis in 3T3-L1 Mature Adipocytes

目的观察山茱萸环烯醚萜苷类提取物对3T3-L1成熟脂肪细胞葡萄糖消耗及异丙肾上腺素(ISO)诱导脂肪分解的影响,探讨其分子机制。方法葡萄糖氧化酶法测定0.04、0.2、1g/L浓度山茱萸提取物干预24h后对脂肪细胞葡萄糖消耗量的影响;比色法测定干预2d后ISO 30min诱导脂肪细胞脂肪分解的甘油释放量;Real-time PCR法检测干预2d后脂肪细胞AMPK、GLUT4和HSL mRNA表达水平,Western blot检测药物对AMPK、ACC蛋白磷酸化的影响。结果 1g/L山茱萸提取物干预成熟脂肪细胞24h后可增加葡萄糖消耗,干预2d后可明显抑制不同浓度ISO诱导的脂肪分解,0.2、0.04g/L提取物可抑制低浓度ISO诱导的脂肪分解;各浓度提取物干预2d可不同程度上调AMPK、GLUT4mRNA水平,下调HSL mRNA表达,1g/L浓度时AMPK、ACC磷酸化水平升高。结论山茱萸提取物可能通过上调AMPK基因表达的相关途径增加3T3-L1脂肪细胞葡萄糖消耗并抑制ISO诱导的脂肪分解。

ABSTRACT：OBJECTIVE To observe the effects of iridoid glycoside extract from Corni Fructus（ECO）on glucose consump-tion and isoproterenol （ISO）-induced lipolysis in 3T3-L1 mature adipocytes and possible mechanism.METHODS Glucose consumption of mature adipocytes intervened by ECO for 24 h was detected in GOP-POD method.After 2 d of ECO interven-tion,colorimetric method was used to evaluate the release amount of glycerol from 30 min ISO-induced lipolysis.The expres-sion of AMPK,GLUT4 and HSL mRNA were detected by Real-time PCR and phosphorylation levels of AMPK and ACC were evaluated by immunoblot.RESULTS In mature 3T3-L1 adipocytes,1 g/L ECO intervention increased glucose consumption and reduced the glycerol release amout from lipolysis induced by ISO with different concentrations.0.2 g/L and 0.04 g/L ECO decreased 100 nmol/L ISO-induced lipolysis.After 2 d intervention,ECO upregulated the levels of AMPK and GLUT4 mRNA and decreased the gene expression of HSL.1 g/L ECO promoted the phosphorylation of AMPK and ACC.CONCLUSION ECO can increase glucose consumption and inhibited ISO-induced lipolysis in 3T3-L1 mature adipocytes and the effects might be related to AMPK regulation.