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目的探讨不同浓度油酸对3T3-L1脂肪细胞脂联素及细胞周期性依赖激酶5(CDK5)、过氧化物酶体增殖物激活受体经(PPARy)表达的调节作用。方法用不同浓度(25、50、100、200、400μmol/L)油酸处理诱导成熟的3T3-L1脂肪细胞24h,用实时荧光定量PCR及Western-blotting法测定各处理组胞内脂联素、CDK5、PPARy mRNA和蛋白表达水平,并在油酸浓度为100、400μmol/L时用Western-blotting法测定PPARγ~(ser273)磷酸化水平。结果50~100μmol/L范围内,油酸可促进脂联素、PPARy mRNA表达,100μmol/L油酸可显著上调脂联素和PPAR;mRNA及蛋白表达;之后随浓度增加,油酸对脂联素和PPARy的促进作用下降,在400μmol/L时显著降低脂联素mRNA和蛋白表达,但对PPARγ蛋白无明显作用。100μmol/L油酸对PPARγ~(ser273)磷酸化水平无明显影响,但在400μmol/L时可显著升高PPARγ~(ser273)磷酸化水平。各浓度油酸对CDK5 mRNA及蛋白均无明显影响。结论50~100μmol/L油酸可促进脂联素、PPARγ表达,400μmol/L油酸抑制脂联素的表达并升高PPARγ~(ser273)磷酸化水平。提示50~100μmol/L油酸可能通过上调PPARγ而促进脂联素表达,400μmol/L油酸可能通过异常激活CDK5介导的PPARγ~(ser273)磷酸化抑制脂联素表达。
Objective To investigate the regulatory effect of different concentrations of oleic acid on the expression of adiponectin, cyclin dependent kinase 5 (CDK5) and peroxisome proliferator activated receptor (PPARy) in 3T3-L1 adipocytes. Methods The mature 3T3-L1 adipocytes were induced with different concentrations of oleic acid (25, 50, 100, 200 and 400μmol / L) for 24 h. The contents of adiponectin and adiponectin in each treatment group were determined by real-time fluorescence quantitative PCR and Western- CDK5 and PPARy mRNA and protein levels were measured. The phosphorylation level of PPARγ ~ (ser273) was determined by Western-blotting at the concentration of 100,400μmol / L oleic acid. Results Oleuric acid could promote the expression of adiponectin and PPARy mRNA in the range of 50 ~ 100μmol / L, and the mRNA and protein expression of adiponectin and PPAR were significantly up-regulated by 100μmol / L oleic acid. With the increase of concentration, The effect of PPARγ and PPARγ decreased. At 400 μmol / L, the mRNA and protein expression of adiponectin decreased significantly, but no significant effect was found on PPARγ protein. 100μmol / L oleate had no significant effect on the PPARγ ~ (ser273) phosphorylation, but at 400μmol / L, PPARγ ~ (ser273) phosphorylation was significantly increased. The concentration of oleic acid had no significant effect on CDK5 mRNA and protein. Conclusions 50 ~ 100μmol / L oleic acid can promote the expression of adiponectin and PPARγ. 400μmol / L oleate inhibited the expression of adiponectin and increased the phosphorylation of PPARγ ~ (ser273). These results suggest that 50-100 micromol / L oleic acid may promote adiponectin expression by up-regulating PPARγ. 400 micromol / L oleic acid may inhibit adiponectin expression through abnormal activation of CDK5-mediated PPARγ- (ser273) phosphorylation.