冠心病痰证患者TGF-β1-PI3K-GAG通路研究  

Study on TGF-β1-PI3K-GAG pathway in patients with phlegm syndrome of coronary heart disease

作　　者：朱璞玉 郑朝阳[2] 丁邦晗[3] 王剑 ZHU Pu-yu;ZHENG Chao-yang;DING Bang-han;WANG Jian(Department of Basic Theory of Chinese Medicine,School of Basic Medical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Departmentl of Cardiology,Guangdong Provincial Hospital of Chinese Medicine,Guangzhou 510000,China;Emergency Department,Guangdong Provincial Hospital of Chinese Medicine,Guangzhou 510000,China)

机构地区：[1]广州中医药大学基础医学院,中西医结合基础研究中心,广州510006 [2]广东省中医院心血管一科,广州510000 [3]广东省中医院急诊科,广州510000

出　　处：《中华中医药杂志》2021年第2期1057-1060,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81873210);广州中医药大学一流学科建设重点项目。

摘　　要：目的:以糖胺聚糖(GAG)为切入点探求冠心病患者痰证相对于阴虚证所存在的差异及其可能机制。方法:收集广东省中医院心血管一科冠心病患者52例,分别提取冠心病痰证及阴虚证患者血清GAG,及患者血清作用小鼠血管平滑肌细胞后的GAG。硫酸咔唑法进行GAG含量测定,凝集素检测法测定GAG种类,Western Blot法测定多能蛋白聚糖Versican,ELISA法测定患者血清中的转化生长因子-β1(TGF-β1)含量和多功能蛋白聚糖Versican含量;TGF-β1依赖Smad通路抑制剂SB525334和PI3K通路抑制剂MK2206干预后,再测定GAG含量,观察痰证与阴虚证GAG检测值差异对应的信号通路。结果:冠心病患者痰证相比于阴虚证,血清中GAG差异无统计学意义。相近血糖水平的痰证患者血清作用于平滑肌细胞后,较阴虚证患者作用后,GAG增高(P<0.05),进一步分析发现增高的GAG主要是硫酸软骨素/硫酸皮肤素(CS/DS),硫酸软骨素类蛋白聚糖(CSPG)中Versican表达升高(P<0.05);与阴虚证比较,痰证患者血清中Versican含量差异无统计学意义,TGF-β1含量增高(P<0.01);添加TGF-β1后,平滑肌细胞GAG含量升高;与阴虚证血清相比,添加痰证血清明显升高平滑肌细胞的GAG含量;在抑制Smad通路后,痰证组GAG比阴虚证组GAG增高的效应仍存在,而在抑制PI3K通路后,痰证组GAG比阴虚证组GAG增高的效应消除了。结论:冠心病痰证患者TGF-β1-PI3K-GAG通路,可能是痰病变的特征信息,其中TGF-β1与生痰因子相关,而GAG和Versican作为痰的可能物质基础,因其主要在组织间隙,在血清中反应不够灵敏。Objective:To explore the difference between phlegm syndrome and yin deficiency syndrome in patients with coronary heart disease(CHD)and its possible mechanism focused on glycosaminoglycan(GAG).Methods:Patients of CHD were recruited from Department I of Cardiology,Guangdong Provincial Hospital of Chinese Medicine.The content of GAGs was determined by Carbazole sufate method,the type of GAGs was determined by lectin assay,Versican was determined by Western Blot,TGF-β1 and Versican in sera of patients were determined by ELISA.Sera that contain Smad depended TGF-β1 pathway inhibitor SB525334 and PI3 K pathway inhibitor MK2206 were applied on MOVAS,then the content of GAGs was measured,and the signaling pathway corresponding to the difference of GAGs content between phlegm syndrome and yin deficiency syndrome was observed.Resuts:There was no significant difference in GAGs content of sera between phlegm syndrome and yin deficiency syndrome.But the content of GAGs increased(P<0.05)after MOVAS cutured with sera of patients with phlegm syndrome when compared with yin deficiency syndrome in the similar level of blood sμgar.Further analysis showed that the main type of increased GAGs was chondroitin sufate/dermatan sufate(CS/DS).Versican increased(P<0.05)after MOVAS cutured with sera of patients with phlegm syndrome when compared with yin deficiency syndrome,and there was no significant difference in the content of Versican in sera of patients between phlegm syndrome and yin deficiency syndrome.With the content of TGF-β1 increased(P<0.01)in sera of patients with phlegm syndrome when compared with yin deficiency syndrome,the content of GAGs also increased after MOVAS treated with recombinant human TGF-β1 when compared with that in normal group.Of course,the content of GAGs increased(P<0.05)after MOVAS cutured with sera of patients with phlegm syndrome when compared with yin deficiency syndrome.GAGs content diminished in both phlegm syndrome and yin deficiency syndrome after inhibiting Smad depended TGF-β1 pathway or PI

关 键 词：硫酸软骨素 多功能蛋白聚糖 转化生长因子-β1 冠心病 痰证 阴虚证 通路 糖胺聚糖 

分 类 号：R259[医药卫生—中西医结合]