过刊浏览

棕榈酸酯诱导人脐带间充质干细胞内质网应激和细胞凋亡

路君, 董会月, 林灵婧, 王庆华, 黄梁浒^*, 谭建明^*

厦门大学附属东方医院(南京军区福州总医院)，福建省移植生物学重点实验室，福州 350025

摘要

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。

关键词： 棕榈酸酯; 间充质干细胞; 细胞凋亡; 内质网应激

分类号：Q256

[Palmitate induces apoptosis and endoplasmic reticulum stress in human umbilical cord-derived mesenchymal stem cells.] [Article in Chinese]

LU Jun, DONG Hui-Yue, LIN Ling-Jing, WANG Qing-Hua, HUANG Liang-Hu^*, TAN Jian-Ming^*

Fujian Provincial Key Laboratory of Transplant Biology, Fuzhou General Hospital, Fuzhou 350025, China

Abstract

The saturated free fatty acid (FFA), palmitate, could induce apoptosis in various cell types, but little is known about its effects on human umbilical cord-derived mesenchymal stem cells (hUC-MSCs). Here, we investigated whether palmitate induced apoptosis and endoplasmic reticulum (ER) stress in hUC-MSCs. hUC-MSCs were stained by labeled antibodies and identified by flow cytometry. After administration with palmitate, apoptotic cell was assessed by flow cytometry using the Annexin V-FITC/7-AAD apoptosis detection kit. Relative spliced XBP1 levels were analyzed using semi-quantitative RT-PCR. The mRNA of BiP, GRP94, ATF4 and CHOP were analyzed by real-time PCR. Relative BiP and CHOP protein were analyzed using Western blot analysis. The results showed that hUC-MSCs were homogeneously positive for MSC markers; palmitate increased apoptosis of hUC-MSCs and activated XBP1 splicing, BiP, GRP94, ATF4 and CHOP transcription. These findings suggest that palmitate induces apoptosis and ER stress in hUC-MSCs.

Key words: palmitate; mesenchymal stem cells; apoptosis; ER stres

收稿日期：2013-04-27　　录用日期：2013-08-05

通讯作者：黄梁浒,谭建明　　E-mail: hlhtiger@yahoo.com.cn,tanjm156@yahoo.com.cn

引用本文：

路君, 董会月, 林灵婧, 王庆华, 黄梁浒, 谭建明. 棕榈酸酯诱导人脐带间充质干细胞内质网应激和细胞凋亡[J]. 生理学报 2013; 65 (6): 600-606.

LU Jun, DONG Hui-Yue, LIN Ling-Jing, WANG Qing-Hua, HUANG Liang-Hu, TAN Jian-Ming. [Palmitate induces apoptosis and endoplasmic reticulum stress in human umbilical cord-derived mesenchymal stem cells.] [Article in Chinese]. Acta Physiol Sin 2013; 65 (6): 600-606 (in Chinese with English abstract).