高血压病血瘀证患者血清对内皮细胞功能及AMPK激活的影响

Effects of the serum of hypertensive patients with blood stasis syndrome on function and AMPK activation of endothelial cells

目的:探索高血压病血瘀证患者血清对内皮细胞功能及AMPK活性的影响.方法:高血压病血瘀证患者血清刺激人脐静脉内皮细胞(HUVECs)24 h,以高血压病非血瘀证患者血清作为对照.采用一氧化氮(NO)、内皮素(ET-1)检测试剂盒检测NO和ET-1的含量.Western blot法检测磷酸化一氧化氮合酶(e NOS)、总e NOS、细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)、磷酸化AMPK及总AMPK的表达.定量PCR法检测肿瘤坏死因子(TNF-α)、白细胞介素-1β(IL-1β)的表达.结果:高血压病血瘀证患者血清处理24 h后,显著降低NO含量、抑制e NOS的活性、升高ET-1的含量,升高ICAM-1、VCAM-1的表达,升高单核细胞黏附率,升高内皮细胞TNF-α、IL-1β的表达水平,与高血压病非血瘀证患者血清相比,有统计学差异(P<0.05).此外,高血压病血瘀证患者血清显著降低蛋白激酶AMPKα的磷酸化水平,与高血压病非血瘀证患者血清相比,有统计学差异(P<0.05).结论:高血压病血瘀证患者血清能诱导内皮功能障碍和炎症反应,其机制可能与抑制AMPK的活性有关.

Objective：This study aimed to investigate the effect of the serum of hypertensive patients with blood stasis syndrome（ BSS） on the function and Adenosine 5＇-monophosphate（ AMP）-activated protein kinase（ AMPK） activation of endothelial cells. Methods： Human umbilical vein endothelial cells（ HUVECs） were treated with the serum of hypertensive patient with BSS for 24 h. The serum of hypertensive patients who are not BSS（ non-BSS） was set as control. The content of nitric oxide（ NO）and endothelin-1（ ET-1） were tested by NO and ET-1 assay kit. Western blot was used to detect the expression of phosphorylated endothelial nitric oxide synthase（ e NOS）, total e NOS, intercellular adhesion molecule 1（ ICAM-1）,vascular cell adhesion molecule 1（ VCAM-1）,phosphorylated AMPK,and total AMPK. Quantitative PCR（ q PCR） was used to detect the expression of tumor necrosis factor-α（TNF-α） and interleukin-1β（IL-1β）. Results： With the treatment of serum of hypertensive patients with BSS,the content of NO was significantly reduced and the activity of e NOS was distinctly inhibited.The level of ET-1,ICAM-1 and VCAM-1 increased,and mononuclear-endothelial cell adhesion rate elevated. The expression of TNF-α and IL-1β was enhanced as well. The results showed significant difference from those of non-BSS group（ P〈 0. 05）. In addition,we also found that the serum of hypertensive patients with BSS significantly reduced the phosphorylation level of AMPKα（ P 〈0. 05）when compared with the results of non-BSS group. Conclusion：The serum of hypertensive patients with BSS can induce endothelial dysfunction and inflammation. Its mechanism may relate to the inhibition of AMPK.