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目的应用基因表达谱芯片技术筛查替米沙坦独立干预下成熟脂肪细胞的差异表达基因,进而对替米沙坦作用下成熟脂肪细胞的功能变化进行预测。方法将3T3-L1前脂肪细胞诱导为成熟脂肪细胞并进行替米沙坦(0.1mg/L)干预,Trizol一步法提取总RNA并纯化,反转录合成荧光分子(Cy3/Cy5)标记的cDNA探针,分别与含有36000个基因或基因片段的高通量小鼠全基因组寡核苷酸微阵列芯片杂交,杂交信号经扫描和数字化处理,筛选替米沙坦干预组与对照组成熟脂肪细胞间的差异表达基因,进而通过生物信息学数据分析推测替米沙坦独立作用下对于成熟脂肪细胞功能的影响以及可能信号途径。结果共筛选157个差异表达基因,其中表达上调的基因86个,表达下调的基因71个,这些基因涉及脂质代谢、细胞分化及成脂过程以及脂肪细胞分泌,其基因功能可能参与替米沙坦对于脂肪细胞功能的独立影响。通过信号通路的生物信息学分析,包括细胞-细胞受体相互作用、细胞黏附分子、脂肪细胞因子信号途径、氧化应激等与脂肪细胞分泌有关的途径均受到影响。此外,替米沙坦还可能作用于脂肪细胞增殖、分化与成脂过程相关途径如Wnt信号途径、β-catenin相关途径以及Notch信号途径等。结论替米沙坦对成熟脂肪细胞可能具有独立于AT受体的特殊作用,并对细胞脂质合成、代谢产生影响。
Objective To screen differentially expressed genes of mature adipocytes under telmisartan independent intervention using gene expression microarray technology and to predict the functional changes of mature adipocytes under the action of telmisartan. Methods 3T3-L1 preadipocytes were induced to mature adipocytes and treated with telmisartan (0.1 mg / L). Total RNA was extracted by one step Trizol and purified. The cDNA labeled with Cy3 / Cy5 was reverse transcribed Probes were hybridized with high-throughput mouse genome-wide oligonucleotide microarray containing 36,000 genes or gene fragments respectively. The hybridization signals were scanned and digitized to screen telmisartan intervention group and control group of mature adipocytes The difference between the expression of genes, and then through bioinformatics analysis of data speculated that telmisartan independent effects on the function of mature adipocytes and the possible signal pathways. Results A total of 157 differentially expressed genes were screened, of which 86 were up-regulated and 71 were down-regulated. These genes involved in lipid metabolism, cell differentiation and adipogenesis, and adipocyte secretion. The gene function may be involved in telmisartan Tan independent effects on fat cell function. Bioinformatics analysis of signal transduction pathways, including cell-cell receptor interactions, cell adhesion molecules, adipocytokine signaling pathways, and oxidative stress, are all involved in adipocyte secretion. In addition, telmisartan may also play an important role in adipocyte proliferation, differentiation and adipogenic processes such as Wnt signaling pathway, beta-catenin pathway and Notch signaling pathway. Conclusion Telmisartan may have special effects on mature adipocytes independent of AT receptors, and affect the lipid synthesis and metabolism of cells.