应用高通量基因表达谱芯片研究替米沙坦对成熟脂肪细胞的影响

Effect of telmisartan on mature adipocytes investigated with microarray based high throughput gene expression profile

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 the differentially expressed genes related to lipid metabolisms in the mature adipo cytes and to investigate the effect of telmisartan on the cells with microarray based high throughput gene expression profile. Methods The 3T3-L1 preadipocytes were induced into the mature adipocytes, which was then intervened with telmisartan （0. 1mg/L）. The total RNA was isolated from the cells with or without telmisartan intervention with Trizol method. After purification, reverse transcription was carried out to synthesize Cy3/Cy5 labeled cDNA probe. The probe was hybridized with microarray based high throughput gene expression profile （36 000 genes or gene fragments） to screen the differentially expressed genes in the cells with and without telmisartan intervention. The effect of telmisartan on the function of the mature adipocytes and the possible signal pathway were analyzed. Results A total of 157 differentially expressed genes were screened out, including 86 up-regulated genes and 71 down-regulated ones, which were involved in lipid synthesis, catabolism and transport in the adipocytes. The changes in the expression level of these genes might be related to the effects of telmisartan on function of mature adipocytes, such as cytokine-cytokine receptor interaction, cell adhesion molecules, adipocytokine signaling pathway, oxidative stress, etc. In addition, telmisartan played some roles in certain pathways related to the proliferation, differentiation, and lipid synthesis of mature adipocytes, such as Wnt, β-catenin, and Notch signaling pathways. Conclusion Telmisartan exerts its effect on mature adipoeytes in a manner which is independent of AT receptor. Telmisartan may affect the lipid synthesis, catabolism and transport of the mature adipocutes.