Study for mechanism of insulin sensitizer using a cDNA subtraction technique

1999年10月使用基因芯片寻找噻唑烷二酮敏感基因胰岛素增敏剂噻唑烷二酮被认为主要通过增强胰岛素对骨骼肌细胞的作用而发挥抗糖尿病作用。PPARg是噻唑烷二酮的靶标，在肝细胞以及脂肪细胞和肌肉细胞中表达。为探讨噻唑烷二酮对小鼠肝细胞基因表达的影响，将小鼠原代肝细胞分别与罗格列酮（5 μmol）和新型噻唑烷二酮化合物MCC 555（25 μmol）孵育，观察噻唑烷二酮对小鼠肝细胞基因表达的影响。培养24小时后，从培养的肝细胞中纯化mRNA。mRNA经荧光标记后与基因芯片杂交。比较对照组、罗格列酮组和MCC55组之间的信号差异，并寻找噻唑烷二酮修饰表达的基因，发现TC20874、TC21818、TC25018、TC26141、TC30984等20多个基因表达水平与对照组相比有显著性差异，其中TC20874、TC21818、TC25018、TC26141和TC30984基因表达水平与对照组相比有显著性差异。 关于我们 用噻唑烷二酮上调ONS。另一方面，包括TC 14248、TC 14302、TC 14653、TC 14842、TC 15141在内的100多个基因被鉴定为噻唑烷二酮下调表达的基因。进一步研究发现，曲格列酮可增加体外培养细胞VEGF的表达，并可增加人血浆VEGF水平，其中包括TC 20874、TC 21818、TC 25018、TC 26141、TC 30984等20多个基因的表达被噻唑烷二酮上调。另一方面，包括TC 14248、TC 14302、TC 14653、TC 14842、TC 15141在内的100多个基因被鉴定为噻唑烷二酮下调表达的基因。此外，还发现曲格列酮可增加培养细胞中VEGF的表达，而处方曲格列酮可增加人血浆VEGF水平。尽管噻唑烷二酮在体外使用细胞系影响单核细胞分化，但当胰岛素增敏剂用于临床时，对白色血细胞的影响尚不清楚。因此，我们试图观察吡格列酮（另一种噻唑烷二酮）临床应用后2型糖尿病患者白色血细胞表面标志物和血清细胞因子水平的变化。我们正在分析数据。少

1999〜Search for thiazolidinedione-sensitive genes using GeneChipInsulin sensitizer, thiazolidinedione, is thought to exert an antidiabetic effect by poteniating the insulin action chiefly on skeletal muscle cells. PPARg, a target of thiazolidinedione, is expressed in hepatocytes as well as adipocytes and muscle cells. To elucidate the mechanism of thiazolidinedione, effects of thiazolidinedione on gene expression of mouse hepatocytes were examined.Mouse primary hepatocytes were incubated in medium in the presence or the absence of 5 μmol of rosiglitazone or 25 μmol of MCC555, a new thiazolidinedione. After 24 hr of culture, mRNA was purified from cultured hepatocytes. mRNA was labelled with fluoressence and was hybridized with GeneChip. Signals were compared among control, rosiglitazone and MCC55, and genes whose expressions were modified with thiazolidinedione were searched.More than 20 genes including TC20874, TC21818, TC25018, TC26141, TC30984 were identified as genes whose expressi … More ons were upregulated with thiazolidinedione. On the other hand, more than 100 genes including TC14248, TC14302, TC14653, TC14842, TC15141 were identified as genes whose expression were downregulated with thiazolidinedione. Furtheremore, it was found that expression of VEGF was increased with troglitazone in cultured cellS and plasma VEGF levels were increased in human with prescribed troglitazone.More than 20 genes including TC20874, TC21818, TC25018, TC26141, TC30984 were identified as genes whose expressions were upregulated with thiazolidinedione. On the other hand, more than 100 genes including TC14248, TC14302, TC14653, TC14842, TC15141 were identified as genes whose expression were downregulated with thiazolidinedione. Furtheremore, it was found that expression of VEGF was increased with troglitazone in cultured cellS and plasma VEGF levels were increased in human with prescribed troglitazone.Though thiazolidinedione was reported to affect monocyte differentiation in vitro using cell lines, nothing is known about the effect on white blood cells when insulin sensitizer is clinically used. We, therefore, attempted to see changes of surface markers on white blood cells and serum cytokine levels from patient with type2 diabetes after pioglitazone, another kind of thiazolidinedione, was clinically used. We are now analyzing data. Less