论文部分内容阅读
饱和游离脂肪酸棕榈酸酯(palmitate,PA)可诱导包括胰岛β细胞在内的多种细胞凋亡,本文旨在观察PA诱导人脐带间充质干细胞(human umbilical cord-derived mesenchymal stem cells,hUC-MSCs)凋亡的作用,并探讨其作用机制。研究采用的hUC-MSCs经细胞表面标志分子抗体标记,通过流式细胞检测鉴定。hUC-MSCs在PA干预后通过Annexin V-FITC和7-AAD双染,流式细胞术检测细胞凋亡率。PA处理后的细胞用于细胞内质网应激标志蛋白检测。半定量PCR观察XBP1 mRNA剪切体,荧光定量PCR检测BiP、GRP94、ATF4和CHOP mRNA表达,Western blot检测BiP和CHOP基因表达。结果显示,培养的hUC-MSCs表达符合MSCs的表面标志分子,PA暴露诱导hUC-MSCs凋亡率显著升高,内质网应激标志基因XBP1 mRNA剪切体增多,BiP、GRP94、ATF4和CHOP基因表达升高。研究结果提示,PA具有诱导hUC-MSCs细胞凋亡的作用,内质网应激可能是细胞凋亡的主要原因。
Saturated free fatty acid palmitate (palmitate, PA) can induce a variety of apoptosis, including islet β cells, the purpose of this paper is to observe PA induced human umbilical cord-derived mesenchymal stem cells (hUC- MSCs) apoptosis, and explore its mechanism of action. The hUC-MSCs used in this study were labeled with cell surface marker antibody and identified by flow cytometry. hUC-MSCs were stained with Annexin V-FITC and 7-AAD after PA intervention, and the apoptosis rate was detected by flow cytometry. PA-treated cells were used for detection of cellular endoplasmic reticulum stress marker protein. The mRNA of BiP, GRP94, ATF4 and CHOP was detected by semi-quantitative PCR and the expression of BiP and CHOP was detected by Western blot. The results showed that the expression of hUC-MSCs in culture was consistent with the surface marker molecules of MSCs, the apoptosis rate of hUC-MSCs induced by PA exposure was significantly increased, the number of XBP1 mRNA endoplasmic reticulum stress gene was increased, and the expression of BiP, GRP94, ATF4 and CHOP Gene expression is elevated. The results suggest that PA has the effect of inducing apoptosis in hUC-MSCs, and endoplasmic reticulum stress may be the main reason of apoptosis.