糖基化终产物对心肌微血管内皮细胞表达MCP-1和ICAM-1的影响及机制研究
Effect and mechanism of AGEs-induced expression of ICAM-1 and MCP-1 on rat cardiac microvascular endothelial cells
赵蓓 1王海昌 1张荣庆 1魏丽萍 1刘毅 1郝媛媛 1尹志勇 1张存1
作者信息
- 1. 第四军医大学西京医院心血管内科,陕西,西安,710033
- 折叠
摘要
目的:观察体外孵育的牛血清白蛋白(BSA)非酶促糖基化终末产物(AGEs)对心肌微血管内皮细胞细胞间黏附分子(ICAM-1)和单核细胞趋化因子-1(MCP-1)表达的影响及机制.方法:取50 g/L牛血清白蛋白(BSA)、500 g/L D-葡萄糖,于37℃孵箱内避光孵育12 wk,制备外源性AGEs-BSA.体外培养大鼠心肌微血管内皮细胞(CMECs).分设不同浓度梯度的AGEs实验组、BSA对照组,采用ELISA法测定MCP-1的表达,流式细胞术测定ICAM-1的表达,Western Blot法测定糖基化终末产物受体蛋白(RAGE)的表达,RT-PCR检测RAGE mRNA的表达.结果:100,200,400 mg/L AGEs可显著增加心肌微血管内皮细胞ICAM-1(13.2%,14.5%,38.1%),MCP-1[(52.5±5.5),(116.0±3.1),(139.6±8.7) μg/L]的表达(与对照组相比较,P<0.05),且在蛋白水平及mRNA水平均明显增加RAGE的表达(P<0.05),并呈浓度依赖性.结论:AGE-BSA可刺激心肌微血管内皮细胞过量表达ICAM-1和MCP-1,从而加速心肌微血管炎症的发生与发展.其机制可能是AGEs上调了心肌微血管内皮细胞RAGE的表达.也进一步证实了AGEs-RAGE信号系统的起动是糖尿病心肌微血管病变发生发展一个重要原因.
Abstract
AIM: To investigate the effect and the mechanism of advanced glycation end products ( AGEs) on the expression of intercellular adhesion molecule-1 ( ICAM-1 ) and monocyte chemotatic protein-1 (MCP-1) in cultured rat cardiac microvascular endothelial cells (CMECs). METHODS: AGEs-BSA was prepared by incubating bovine serum albumin ( BSA) with high concentration of D-glucose at 37℃ in vitro. CMECs were cultured according to the descriptions in literatures. The production of ICAM-1 induced by AGEs was determined by flow cytometry (FCM) and MCP-1 was examined by enzyme-linked immunosorb-nent assay ( ELISA). Receptor for advanced glycation end products (RAGE) was detected at both protein and mRNA levels using Western blot and RT-PCR,respectively. RESULTS: AGEs (100,200 and 400 mg/L) stimulated CMECs to over-express MCP-1 [(52.5±5.5),(116.0±3.1),(139.6 ±8.7)μg /L] and ICAM-1 ( 13. 2%,14. 5%,38. 1% ) and increased the expression of RAGE at both protein and mRNA levels in a dose concentration-dependent manner (all P<0.05). CONCLUSION: AGEs up-regulates ICAM-1 and MCP-1 expressions and increases expression of RAGE in CMEC and the underlying mechanism may be that AGEs increase the expression of proinflammatory cytokines through up-regulation of RAGE. These findings further suggest that AGE-RAGE interaction is a key contributing factor leading to the development of microvascular diabetic complications.
关键词
糖基化终产物,高级/耱基化终末产物受体/心肌/微血管/内皮细胞Key words
glycosylation end products,advanced/receptor for advanced glycosylation end products/myocardium/microvascular/endothelial cells引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据