内质网应激在尿毒症血清致内皮细胞功能异常中的作用  

Effect of endoplasmic reticulum stress on uremic serum-induced endothelial cell dysfunction

作　　者：曾薇[1] 罗志锋[1] 齐伟[1] 郭艳红[1] 庞琦[1] 张均[1] 王宗前[1] 牟娇[1] 冯兵[1] 

机构地区：[1]第三军医大学新桥医院肾内科,重庆400037

出　　处：《解放军医学杂志》2011年第2期133-136,共4页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(30570763)

摘　　要：目的探讨内质网应激(ERS)在尿毒症血清致内皮细胞功能异常中的作用。方法采集尿毒症患者和正常健康人的静脉血血清。以不同浓度的尿毒症血清或正常血清(2.5%、5%、10%、15%、20%)刺激人脐静脉内皮细胞(hUVEC)24h后,采用MTT比色法检测细胞活力的变化,流式细胞术检测细胞周期,Hoechst33342染色观察细胞凋亡,荧光定量RT-PCR法检测单核细胞趋化蛋白-1(MCP-1)和葡萄糖调节蛋白78(GRP78)mRNA表达的变化。结果当浓度小于10%时,尿毒症血清对hUVEC细胞增殖率和S期百分比的影响随浓度增加而增大。当浓度大于10%时,hUVEC细胞增殖率和S期百分比逐渐下降,且细胞凋亡率逐渐增高。与正常血清组比较,10%尿毒症血清刺激后hUVEC GRP78和MCP-1 mRNA表达均显著升高(P<0.05)。应用分子伴侣4-苯基丁酸(4-PBA)干预,可显著抑制尿毒症血清诱导的GRP78 mRNA和MCP-1 mRNA表达(P<0.05),降低hUVEC的增殖率和S期百分比(P<0.05)。结论内质网应激可能是尿毒症血清诱导内皮细胞功能异常的重要细胞内机制。Objective To investigate the effect of endoplasmic reticulum stress in uremic serum-induced endothelial cell dysfunction.Methods Human umbilical vein endothelial cells（hUVECs） were cultured in growth medium with the addition of uremic serum collected from uremic patients in different concentrations,or normal serum collected from normal adults for 24h.The cell viability was then tested by MTT assay and the cell cycle was analyzed by flow cytometry（FCM）.Hoechst33342 assay was used to analyze the apoptotic effect of uremic serum on hUVECs.The cellular mRNA expressions of monocyte chemotactic protein-1（MCP-1） and glucose-regulated protein 78（GRP78） were measured by real-time fluorescence PCR.Results Co-cultured with lower concentrations of uremic serum（〈10%）,the proliferation and percentage of S stage of cell cycle in hUVECs increased gradually and significantly.In contrast,the proliferation of hUVECs decreased gradually and the apoptosis of hUVECs increased when cultured with higher concentrations（〉10%） of uremic serum.Expressions of GRP78 and MCP-1 mRNA increased significantly in 10% uremic serum than in normal serum（P〈0.05）.However,induction of expressions of GRP78 and MCP-1 mRNA by uremic serum could be inhibited significantly by the chemical chaperone 4-phenylbutyric acid（4-PBA）（P〈0.05）.Furthermore,cell viability and the percentage of S stage of hUVECs were decreased（P〈0.05）.Conclusion Endoplasmic reticulum stress plays a critical role in uremic serum-induced endothelial cell dysfunction.

关 键 词：内质网应激 4-苯基丁酸 尿毒症 人脐静脉内皮细胞 动脉粥样硬化 

分 类 号：R692.5[医药卫生—泌尿科学]