摘要
目的分离和克隆肿瘤坏死因子α(TNF α)致 3T3 L1脂肪细胞胰岛素抵抗 (IR)的相关基因。方法分化成熟的 3T3 L1脂肪细胞用TNF α处理 4 8h后抽提总RNA ,采用mRNA差异显示技术分离和克隆TNF α引起脂肪细胞IR的基因 ,再通过半定量RT PCR证实。结果经mRNA差异显示技术分离和克隆到 10个已知基因的cDNA ,3个已知表达序列标签 (ESTs)片段和1个新的EST片段。半定量RT PCR证实TNF α上调 3T3 L1脂肪细胞Synip基因和血浆淀粉样蛋白A3(SAA3)基因的表达。结论Synip基因和SAA3基因的表达升高可能与TNF α致 3T3 L1脂肪细胞IR和相关的心血管并发症有关。
Objective To isolate and clone the genes related to TNF-α-induced insulin resistance in 3T3-L1 adipocytes. Methods After 3T3-L1 adipocytes treated with TNF-α for 48 h, the mRNA differential display method was used to compare and analyze mRNAs prepared from 3T3-L1 adipocytes and control cells and semi-quantitative RT-PCR was used to confirm the differentially expressed genes found by mRNA differential display method. Results A total fourteen cDNA fragments differentially expressed in 3T3-L1 adipocytes treated with TNF-α were identified. Subsequent cloning and sequencing of these cDNA fragments and BLASTN analysis resulted in the identification of 10 known genes, three known ESTs and one unknown EST. The Synip and serum amyliod protein A3(SAA3) genes were confirmed to be up-regulated in 3T3-L1 adipocytes treated with TNF-α using semi-quantitative RT-PCR. Conclusion Up-regulation of the Synip and SAA3 genes may be related to TNF-α-induced insulin resistance in 3T3-L1 adipocytes and cardiovascular complications.
出处
《上海第二医科大学学报》
CSCD
2004年第12期997-1000,共4页
Acta Universitatis Medicinalis Secondae Shanghai
基金
国家科技攻关计划 (2 0 0 2BA71 1A0 5)资助项目
