摘要
目的旨在观察补脏通络方含药血清对高糖环境下人脐静脉内皮细胞(HUVEC)NO、eNOS及MMP9/TIMP1表达的影响,探讨补脏通络方保护糖尿病大血管内皮的可能机制。方法采用血清药理学方法提取补脏通络方含药血清。体外培养HUVEC,细胞分正常组、高糖组、补脏通络方低剂量含药血清干预组(BZTL-L)、补脏通络方中剂量含药血清干预组(BZTL-M),补脏通络方高剂量含药血清干预组(BZTL-H)进行干预实验。硝酸还原酶法检测细胞上清液NO水平,Western Blot分析细胞eNOS、MMP9、TIMP1蛋白含量。结果高糖组较正常组,NO水平和eNOS、TIMP-1表达均降低(P<0.01),MMP-9表达和MMP-9/TIMP-1比值均升高(P<0.01);各中药干预组较高糖组,NO水平和eNOS、TIMP-1表达均升高(P<0.01),MMP-9表达和MMP-9/TIMP-1比值均下降(P<0.01),且MMP-9/TIMP-1比值下降呈剂量依赖性。结论改善内皮细胞NO分泌,调节内皮细胞MMP-9/TIMP-1平衡以稳定血管基质的正常代谢,可能是补脏通络方保护糖尿病大血管内皮的具体机制之一。
Objective To study the Effect of Bu Zang Tongluo Decoction medicated serum interferred on NO,e NOS,MMP-9 /TIMP-1 in Human umbilical vein endothelial cells（ HUVEC） under high sugar and discuss the mechanism of Bu Zang Tongluo Decoction intervening diabetic large vascular disease. Methods Extract Bu Zang Tong Iuo Decoction medicated serum with the method of serum pharmacology. HUVEC cultivated in vitro were divided into normal group,high sugar group and Bu Zang Tong Iuo Decoction medicated serum low-dose group（ BZTL-L）,Bu Zang Tongluo Decoction medicated serum middle-dose group（ BZTL-M）,Bu Zang Tong Iuo Decoction medicated serum high-dose group（ BZTL-H） to do intervening experiment. The content of NO in supernatant of cell was tested by Nitrate reductive enzymatic and the expression of e NOS,MMP9,TIMP1 in cell were tested by Western Blot. Results Compared with normal group,the content of NO and the expression of e NOS,TIMP-1 were reduced（ P〈0. 01） and the expression of MMP-9 and the ratio of MMP-9 / TIMP-1 were increased in high sugar group（ P〈0. 01）.Compared with high sugar group,the content of NO and the expression of e NOS,TIMP-1 were increased（ P〈0. 01）,the expression of MMP-9 and the ratio of MMP-9 / TIMP-1 were decreased（ P〈0. 01） in each herb intervening group and the ratio of MMP-9 / TIMP-1 showed dose-dependent. Conclusion Improving the secretion of NO in endothelial cells and regulating MMP-9 / TIMP-1 to stabilize the metabolism of vascular matrix may be relate to the protective mechanism of Bu Zang Tongluo Decoction on diabetic vascular endothelium.
出处
《时珍国医国药》
CAS
CSCD
北大核心
2015年第12期2832-2834,共3页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金青年基金(No.81202733)
湖北省武汉市科技局科技攻关计划(武科计【2013】20号)
湖北中医药大学校级重点项目(2013[144]号)
