丹参对高血压病血瘀证血清诱导内皮功能障碍的影响

Effects of Salvia Miltiorrhiza on Endothelial Dysfunction Induced by Serum of Hypertension Patients with Blood Stasis Syndrome

目的:探索丹参对高血压病血瘀证患者血清诱导的内皮功能障碍及腺苷酸活化蛋白激酶(AMPK)活性的影响,并进一步探讨丹参保护内皮功能障碍的潜在作用机制。方法:以人脐静脉内皮细胞(HUVEC)为研究对象,利用高血压病血瘀证患者血清诱导内皮功能障碍,用不同浓度的丹参(50、100μg/mL)处理24 h后,采用试剂盒检测一氧化氮(NO)及内皮素(ET-1)的含量。采用Western Blot检测磷酸化及总的一氧化氮合酶(eNOS)、细胞间黏附分子-1 (ICAM-1)、血管细胞黏附分子-1(VCAM-1)、磷酸化及总的AMPK的表达。采用荧光定量PCR技术检测肿瘤坏死因子(TNF-α)、白细胞介素-1β(IL-1β)的表达。结果:丹参能剂量依赖性的增加NO的含量、降低ET-1的含量、增加eNOS的磷酸化水平,降低内皮细胞黏附分子ICAM-1、VCAM-1的表达,减少单核细胞黏附率,降低炎性细胞因子TNF-α、IL-1β水平,升高AMPK磷酸化水平,与模型组比较差异均具有统计学意义(P<0.05)。结论:丹参能够改善高血压病血瘀证患者血清诱导的内皮功能障碍,其机制可能与激活AMPK有关。

Objective: To explore the effect of salvia miltiorrhiza on endothelial dysfunction induced by serum of hypertensive patients with blood stasis syndrome and Adenosine 5'-monophosphate( AMP)-activated protein kinase activity and further explore the potential mechanisms in such effects. Methods: The human umbilical vein endothelial cells ( HUVECs) were used in this research. The blood serum of hypertensive patients with blood stasis syndrome was used to induce endothelial dysfunction and then it was treated with different concentrations of Salvia miltiorrhiza (50, 100 μg/mL) for 24 h. Kits were used to detect the levels of nitric oxide( NO) and endothelin( ET - 1). Phosphorylated and total endothelial nitric oxide synthase(eNOS), intercellular adhesion molecule - 1 (ICAM - 1), vascular cell adhesion molecule -1 ( VCAM - 1), phosphorylated and total AMPK were detected by Western Blot. The expressions of tumor necrosis factor -a( TNF -α) and interleukin - 1 p( IL - 1β) were detected by real - time PCR. Results:Salvia miltiorrhiza can dose -dependently increase the amount of NO and the phosphorylation level of eNOS, while it decreased the amount of ET-1 , the expressions of ICAM-1 and VCAM - 1 and reduced the adhesion rate of monocytes. Furthermore, the levels of inflammatory cytokines TNF -α and IL-10 were decreased and the phosphorylation level of AMPK was increased. All data was statistically significant compared with those of the model group( P < 0. 05 ). Conclusion: Endothelial dysfunction induced by serum of hypertensive patients with blood stasis syndrome was significantly improved by Salvia miltiorrhiza and its mechanism may be related to the activation of AMPK.