过表达SCD1对人骨髓间充质干细胞成内皮分化的影响以及全基因组表达谱变化研究

Effect of SCD1 Overexpression on the Endothelial Differentiation Function of Bone Marrow Mesenchymal Stem Cells and Differential Gene Expression Profile

目的:探讨过表达固醇辅酶A去饱和酶1(SCD1)对人骨髓间充质干细胞(BM-MSCs)成内皮作用的影响,并通过基因芯片及智能通路(IPA)分析系统研究全基因组表达谱变化。方法:利用已构建成功的SCD1慢病毒转染BM-MSCs,采用RT-PCR及C14技术检测SCD1在BM-MSCs中过表达情况及其活性。内皮诱导培养BM-MSCs后,采用RT-PCR技术检测CD31、v WF及CDH5等相关内皮指标,进一步运用全基因芯片检测SCD1过表达对BM-MSCs成内皮分化表达谱的影响。结果:BM-MSCs成功过表达SCD1并保持高活性。内皮诱导培养7天时,过表达组的内皮指标CD31、v WF m RNA高于对照组(p<0.05)、14天时过表达组的CD31、v WF及CDH5 m RNA均高于对照组(p<0.05)。基因芯片结果显示SCD1改变BM-MSCs内皮分化表达谱,共有522个差异基因被检测出。IPA结果显示Nrf2通路及细胞分化功能的表达差异显著(p<0.05)。结论:SCD1过表达可以促进BM-MSCs的成内皮分化,可能通过降低细胞氧化应激、提高细胞增殖分化能力实现。SCD1这种抗氧化作用可能为内皮功能修复及心血管疾病治疗提供潜在的策略,值得深入研究。

Objective： Explored the effect of Stearoyl-Co A desaturase 1（SCD1） overexpression on the endothelial differentiation function of human bone marrow mesenchymal stem cells（BM-MSCs） and genes expression profile, through the gene chip and intelligent pathway analysis system（IPA）. Methods： Used the SCD1 lentivirus which had been successfully built to transfect BM-MSCs. Performed RT-PCR to detect the expression of SCD1 and used ratio of（[14 C] oleic acid）/（[14 C] oleic and stearic acids） to detected the activity of SCD1. After endothelial induction culture, used RT-PCR to detect the endothelial index including CD31, v WF and CDH5. Gene microarray was performed and used the Ingenuity Pathway Analysis（IPA） system to analyze the differential gene expression profile of the BM-MSCs modulated by SCD1 overexpression. Results： SCD1 was stably overexpressed in BM-MSCs and maintained a highly activity. After 7 days of endothelial induction culture, CD31 and v WF m RNA of overexpression group were higher than that of the control group（p〈0.05）, and CD31, v WF and CDH5 m RNA of overexpression group were higher than that of the control group after 14 days（p〈0.05）. The microarray showed there are total 522 differential expression genes and IPA results show that the genes of NRF2 pathway and the genes of Cell Differentiation Function were significantly differential expression genes（p〈0.05）. Conclusion： SCD1 expression can promote the endothelial differentiation of BM-MSCs, probably by reducing the cell oxidative stress and improving the ability of cell proliferation differentiation. This effect of SCD1 may provide the potential strategy for the endothelial function repair and cardiovascular diseases treatment, which is worth further study.