糖基化终末产物对3T3-L1脂肪细胞脂联素分泌的影响

EFFECT OF ADVANCED GLYCATION END PRODUCTS ON THE EXPRESSION OF ADIPONECTIN IN 3T3-L1 ADIPOCYTES

目的糖基化终末产物(Advanced glycation end products,AGE)对3T3-L1脂肪细胞脂联素(adiponectin,APN)分泌的影响。方法3T3-L1小鼠前脂肪细胞体外培养并诱导分化为成熟的脂肪细胞,以PBS和BSA作为阴性对照,用含不同浓度梯度(50μg/ml、100μg/ml、150μg/ml)AGE的培养液对成熟的脂肪细胞体外培养48h,收集细胞和上清液,用RT-PCR方法检测脂联素mRNA的表达,ELISA方法检测培养液中脂联素蛋白的分泌情况。结果AGE干预组脂联素的表达在mRNA和蛋白水平较对照组均明显下降(P<0.05),并且随着AGE浓度的升高脂联素的合成和分泌逐渐减少,其中100μg/ml、150μg/ml AGE干预组脂联素的减少较对照组有显著差异(P<0.001)。结论糖基化终末产物能够通过抑制3T3-L1细胞脂联素mRNA的合成,近而抑制脂联素的分泌,且这种抑制呈浓度依赖性。

Objective To explore the effect of advanced glycation end products on the expression of adiponectin in mouse adipocytes. Methods 3T3-L1 adipocytes were cultured and differentiated in vitro into mature adipocytes. These cells were incubated for 48 hours in the presence of 50μg/ml, 100μg/ml or 150μg/ml AGE-BSA, followed by assay for adiponectin synthesis and secretion. We established 2 control groups with unmodified BSA and phosphate-buffered saline （PBS）. Adiponectin mRNA was quantitated by RT-PCR, while adiponectin concentrations in the culture medium were determined by ELISA. Results Adiponectin expression of differentiated adipocytes incubated with AGE-BSA was inhibited significantly as compared with that of the control groups （P〈0. 05）. Along with the increasing AGE concentration, adiponectin expression was gradually decreasing. There were significant differences between the 100μg/ml or 150μg/ml AGE experimental groups and the control groups （P〈0. 001）. Conclusion These results clarify that AGE-BSA is able to inhibit adiponectin synthesis at a mRNA level, leading to reduction in adiponectin secretion from these adipocytes, and this inhibition is dose-dependent.