黄连多糖对AGEs诱导内皮细胞增殖及其受体表达的作用研究  

Effects of Polysaccharides from Coptis Chinensis on HUVECs Proliferation Induced by Advanced Glycation Endproducts and Expression of Its Receptor

作　　者：尹登科[1] 杨晔[1] 陈松[2] 李云[1] 高向东[2] 

机构地区：[1]安徽中医学院药学院,安徽合肥230031 [2]中国药科大学生命科学与技术学院,江苏南京210009

出　　处：《药物生物技术》2012年第6期476-479,共4页Pharmaceutical Biotechnology

基　　金：天然药物活性组分与功效国家重点实验室(中国药科大学)资助项目(SKLNMKF201218);安徽省高校优秀人才基金(No.2011SQRL091)

摘　　要：考察黄连多糖对高级糖基化终产物(AGEs)诱导人脐静脉内皮细胞(HUVECs)增殖和AGEs受体(RAGE)表达的作用。采用水提,Sevag法去蛋白,醇沉法获得黄连多糖(CCP);80%汇聚的HUVECs分成6组,分别为空白对照组、BSA对照组(蛋白浓度200μg/mL)、AGEs组(蛋白浓度200μg/mL)、AGEs+CCP(25μg/mL)、AGEs+CCP(50μg/mL)和AGEs+CCP(100μg/mL),采用MTT法检测黄连多糖对AGEs诱导HUVECs增殖的影响;实时荧光定量PCR检测RAGE mRNA表达;Western Blot分析RAGE蛋白表达情况。HUVECs经AGEs诱导48h后,其增殖率显著增殖。黄连多糖可以剂量依赖性的抑制AGEs诱导HUVECs早期增殖作用,定量PCR和Western Blot结果表明CCP可以在mRNA和蛋白水平抑制RAGE表达。黄连多糖可通过抑制RAGE表达,降低AGEs对内皮细胞的激活作用。To study the effects of Coptis Chinensis polysaccharide （CCP） on HUVECs proliferation induced by advanced glycation endproducts （AGEs） and the expression of the receptor for AGEs （RAGE）, the total CCP was prepared by water extraction, depro- teinized by method of sevag, and alcohol precipitation. HUVECs with 80% confluent were divided into six groups as control（ without treatment）, BSA group （ 200 μg/mL ）, AGEs group （ 200 μg/mL, protein concentration ）, AGEs ＋ CCP （ 25 μg/mL）, AGEs ＋ CCP （50μg/mL） and AGEs ＋ CCP（ 100 μg/mL）, The proliferation of HUVECs was determined by the method of MTY, Real Time Quantitative Fluorescence RCR was used to analyze the expression of RAGE mRNA and Western Blot was used to detect the expression of RAGE. The proliferation of HUVECs was increased after treatment with AGEs for 48 h, CCP significantly inhibited the pro-proliferation of HUVECs induced by AGEs in dose-dependent manner. The results of PCR and Western Blot also demonstrated that CCP could decrease the expression of RAGE mRNA and protein. CCP inhibited the activation of HUVECs induced by AGEs through inhibiting the expression of RAGE.

关 键 词：黄连多糖 高级糖基化终产物 内皮细胞 增殖 受体 实时定量PCR 

分 类 号：Q539[生物学—生物化学]