Department of Orthopedics, Shanghai First People's Hospital, School of Medicine, Shanghai Jiao Tong University, 100, Haining Road, Shanghai 200080, China.
Lipids Health Dis. 2014 Mar 20;13:53. doi: 10.1186/1476-511X-13-53.
Bone marrow mesenchymal stem cells (BM-MSCs) are capable of differentiating into endothelial cells in vitro and acquire major characteristics of mature endothelial-like expression of vWF and CD31. SFAs and lipid oxidation products have been linked with postprandial endothelial dysfunction. Consumption of SFAs impairs arterial endothelial function, while a Mediterranean-type MUFA-diet has a beneficial effect on endothelial function by producing a decrease in levels of vWF, TFPI and PAI-1. Stearoyl-CoA desaturase 1 (SCD1), which converts SFA to MUFA, is involved in the cellular biosynthesis of MUFAs from SFA substrates. High expression of SCD1 is corresponded with low rates of fatty acid oxidation, therefore it might reduce inflammatory responses and be beneficial for the growth of induced endothelial cells. Overexpression of SCD1 in BM-MSCs might increase the growth of induced endothelial cells. The goal of this research is to study the relationship between overexpression of SCD1 and the expression of induced endothelial cells in BM-MSCs in vitro.
The gene SCD1 was integrated into a lentiviral vector, and then 293 T cells were transfected by the connected product to produce a packaged virus. BM-MSCs were infected by the packaged virus. Cell culture and endothelial induction were performed. Fluorescent quantitative PCR of CD31, vWF and VE-cad was performed after 1 week and 2 weeks to test the growth of induced endothelial cells.
The mRNA amount of CD31, vWF and VE-cad of the SCD1 overexpressed group was statistically higher than that of the empty vector (EV) group and that of the normal group after 1 week and 2 weeks, respectively (p < 0.05). Immunocytochemical staining of CD31 or vWF was detected by visualizing red color.
This study suggested that overexpression of SCD1 in BM-MSCs could increase the expression of induced endothelial cells in vitro.
骨髓间充质干细胞（BM-MSCs）在体外可分化为内皮细胞，并获得 vWF 和 CD31 等成熟内皮样表达的主要特征。饱和脂肪酸（SFAs）和脂质氧化产物与餐后内皮功能障碍有关。SFAs 的消耗会损害动脉内皮功能，而地中海型 MUFA 饮食通过降低 vWF、TFPI 和 PAI-1 的水平对内皮功能产生有益影响。将 SFA 转化为 MUFA 的硬脂酰辅酶 A 去饱和酶 1（SCD1）参与了从 SFA 底物到 MUFA 的细胞生物合成。SCD1 高表达与脂肪酸氧化率低相关，因此它可能会降低炎症反应，有利于诱导内皮细胞的生长。在 BM-MSCs 中超表达 SCD1 可能会增加诱导的内皮细胞的生长。本研究旨在研究 SCD1 过表达与体外 BM-MSCs 中诱导的内皮细胞表达之间的关系。
将 SCD1 基因整合到慢病毒载体中，然后将连接产物转染 293 T 细胞以产生包装病毒。BM-MSCs 被包装病毒感染。进行细胞培养和内皮诱导。在第 1 周和第 2 周时，通过荧光定量 PCR 检测 CD31、vWF 和 VE-cad 的表达，以检测诱导的内皮细胞的生长。
SCD1 过表达组在第 1 周和第 2 周时，CD31、vWF 和 VE-cad 的 mRNA 量分别高于空载体（EV）组和正常组（p<0.05）。通过可视化红色来检测 CD31 或 vWF 的免疫细胞化学染色。
本研究表明，在 BM-MSCs 中超表达 SCD1 可以增加体外诱导的内皮细胞的表达。
