脂肪因子Vaspin体外表达与妊娠期糖尿病胰岛素抵抗研究

Effect of Vaspin in vitro expressed on insulin signal transduction pathway of GDM

目的:探讨脂肪因子Vaspin与妊娠期糖尿病(GDM)胰岛素抵抗的内在联系。方法:复苏、传代及诱导分化GDM前脂肪细胞,构建Vaspin过表达载体,转化、培养和提取;以4个过表达梯度(0.0、1.0、2.5、5.0μg)转染传代脂肪细胞,Q-PCR检测Vaspin、胰岛素受体底物-1/2(IRS-1/2)、磷脂酰肌醇-3激酶(PI3K(P85a))m RNA表达,WB检测Vaspin、IRS-1/2、PI3K(P85a)蛋白表达及IRS-1/2磷酸化水平,[3H]-2-脱氧-D-葡萄糖摄取测定法检测各组葡萄糖摄取率的变化。结果:(1)Vaspin m RNA及蛋白表达随转染浓度梯度升高而表达量增加,所构建Vaspin过表达载体有效;(2)随Vaspin表达增加,IRS-1/2、PI3K(P85a)m RNA、蛋白表达及IRS-1/2酪氨酸磷酸化程度均未出现明显变化;(3)脂肪细胞葡萄糖摄取率与Vaspin过表达浓度变化无明显关系。结论:Vaspin在GDM血清及网膜脂肪组织中的高表达与GDM胰岛素抵抗的发生没有必然联系。

Objective To investigate the relationship between Vaspin and GDM insulin resistance. Methods The GDM preadipocyte cells were recoveried, extended and differentiated.Vaspin overexpression carriers were made to transformate, cultivate and extract the plasmid. The fat cells were transfected and extended using 4 overexpression levels （0.0 μg, 1.0 μg, 2.5 μg, 5.0 μg）. Q-PCR was used to detect mRNA expression of Vaspin, insulin receptor substrate-1/2 （IRS-1/2）, phosphatidy inositol 3 kinase （PI3K （P85a） Western Blot was used to detect protein expression of Vaspin, IRS-1/2, PI3K （P85a） and IRS-1/2 phosphorylation levels, and [3H]-2-deoxidation-D-glucose uptake assay was used to detecte glucose uptake rates. Results （1） According to the Q-PCR and WB results, the constructed Vaspin overexpression carrier was effective; （2） With the Vaspin expression increased, the mRNA and protein expression of IRS-1/2, PI3K （P85a） and IRS-1/2 tyrosine phosphorylation levels had no significant changes;（3） Glucose uptake rate of fat cells had no obvious correlation with Vaspin. Conclusion High expression of Vaspin in GDM serum and omental adipose tissue has no obvious link with the insulin resistance of GDM.