High glucose mediates endothelial-to-chondrocyte transition in human aortic endothelial cells
High glucose mediates endothelial-to-chondrocyte transition in human aortic endothelial cells
复制标题
高葡萄糖介导人主动脉内皮细胞内皮细胞向软骨细胞的转变
DOI：
10.1186/1475-2840-11-113
复制
发表时间：
2012-09-22
影响因子：
9.3
通讯作者：
Liu, Bicheng
中科院分区：
文献类型：
--
作者：
Tang, Rining;Gao, Min;Liu, Bicheng
背景：血管钙化是糖尿病常见的并发症之一。许多研究表明，高糖(HG)导致心血管钙化，但其潜在机制尚不完全清楚。最近，糖尿病患者的血管内膜钙化是最常见的。软骨细胞参与内侧钙化。最近的研究表明，通过内皮细胞向间充质干细胞的转化(EndMT)可以在软骨细胞中触发向间充质干细胞(MSCs)的转化。我们先前的研究表明，HG可诱导人主动脉内皮细胞(HAECs)发生EndMT。方法：将人脐静脉内皮细胞分为3组：正常葡萄糖(NG)组、HG组(30 mm o l/L)和甘露醇组(5.5 m o l/L NG+24.5 m o l/L)。用荧光显微镜和电子显微镜观察病理变化。免疫荧光染色检测CD31等内皮细胞标志物与成纤维细胞特异性蛋白1(FSP-1)共表达情况。实时荧光定量聚合酶链式反应和免疫印迹法检测FSP-1的表达。将内皮源性MSCs在MSC培养液中培养一周。免疫荧光染色和免疫印迹法检测MSCs标志物Stro-1、CD44、CD10和软骨细胞标志物SOX9的表达。阿新蓝染色检测软骨细胞的表达。结果：HG孵育HAECs后，细胞出现成纤维细胞样表型。HAECs双重染色显示CD31和FSP-1共存。HG组FSP-1的表达显著增加，EndMT组细胞也表达Stro-1、CD44和SOX9，与对照组相比差异有统计学意义(P<；0.05)。此外，HG组阿尔新蓝染色阳性，而NG组则为阳性。与SOX9表达的评估一致，茜素红染色分析的钙沉积也被HG处理增强。结论：HG可通过EndMT诱导内皮细胞转分化为软骨细胞样细胞，这一过程部分是通过激活Snail信号通路实现的。
Background: Vascular calcification is one of the common complications in diabetes mellitus. Many studies have shown that high glucose (HG) caused cardiovascular calcification, but its underlying mechanism is not fully understood. Recently, medial calcification has been most commonly described in the vessels of patients with diabetes. Chondrocytes were involved in the medial calcification. Recent studies have shown that the conversion into mesenchymal stem cells (MSCs) via the endothelial-to-mesenchymal transition (EndMT) could be triggered in chondrocytes. Our previous research has indicated that HG induced EndMT in human aortic endothelial cells (HAECs). Therefore, we addressed the question of whether HG-induced EndMT could be transitioned into MSCs and differentiated into chondrocytes.Methods: HAECs were divided into three groups: a normal glucose (NG) group, HG group (30 mmol/L), and mannitol (5.5 mmol/L NG + 24.5 mmol/L) group. Pathological changes were investigated using fluorescence microscopy and electron microscopy. Immunofluorescence staining was performed to detect the co-expression of endothelial markers, such as CD31, and fibroblast markers, such as fibroblast-specific protein 1 (FSP-1). The expression of FSP-1 was detected by real time-PCR and western blots. Endothelial-derived MSCs were grown in MSC medium for one week. The expression of the MSCs markers STRO-1, CD44, CD10 and the chondrocyte marker SOX9 was detected by immunofluorescence staining and western blots. Chondrocyte expression was detected by alcian blue staining. Calcium deposits were analyzed by alizarin red staining.Results: The incubation of HAECs exposed to HG resulted in a fibroblast-like phenotype. Double staining of the HAECs indicated a co-localization of CD31 and FSP-1. The expression of FSP-1 was significantly increased in the HG group, and the cells undergoing EndMT also expressed STRO-1, CD44 and SOX9 compared with the controls (P < 0.05). Additionally, alcian blue staining in the HG group was positive compared to the NG group. Consistent with the evaluation of SOX9 expression, calcium deposits analyzed by alizarin red staining were also enhanced by the HG treatment. Specifically, we showed that HG-induced EndMT is accompanied by the activation of the canonical Snail pathway.Conclusions: Our study demonstrated that HG could induce endothelial cells transdifferentiation into chondrocyte-like cells via the EndMT, which is mediated in part by the activation of the Snail signaling pathway.