川芎嗪对脂多糖诱导的血管内皮细胞损伤的影响

目的探讨川芎嗪(tetramethylpyrazine,TMP)对脂多糖(lipopolysaccharide,LPS)诱导引起的人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)损伤的保护作用。方法以人脐静脉内皮细胞为实验对象,MTT法检测细胞活力,比色法测定一氧化氮合酶(nitric oxide synthase,NOS)活性,硝酸还原酶法测上清液一氧化氮(nitric oxide,NO)含量,荧光法测定胞内NO及ROS水平,流式细胞仪法测定细胞凋亡与周期。结果川芎嗪与脂多糖同内皮细胞孵育后没有引起细胞活力的明显改变,排除了实验模型中药物直接细胞毒作用。脂多糖可以抑制细胞NOS活性(P<0.05)及胞内外NO含量(P<0.05),增加胞内ROS水平(P<0.01),促进细胞凋亡(P<0.05),抑制正常细胞周期的进程(P<0.05),而川芎嗪可以恢复脂多糖所致的上述效应,并具有一定的浓度依赖性。结论川芎嗪可以通过有效抑制脂多糖诱导的HUVEC NO、ROS水平改变所引起的细胞凋亡,从而起到内皮细胞损伤的保护作用。

Effect of tetramethylpyrazine on lipopolysaccharide-induced damage in vascular endothelial cell

Aim To investigate the protective effect of Tetramethylpyrazine (TMP) on damage in human umbilical vein endothelial cells (HUVECs) induced by Lipopolysaccharide (LPS).Method HUVECs were tested by MTT assay for cell viability, colorimetric method for activity of nitric oxide synthase (NOS), measurement for supernatant of nitric oxide(NO) with nitrate reductase method. The level of intracellular NO and reactive oxygen species (ROS) were estimated through fluorescence assay. The cell apoptosis and cycle were analyzed on a FACS Vantage SE flow cytometer.Results The cell viability had no obviously influence by TMP pre-incubated with or without LPS (P>0.05). It was established that both TMP and LPS had less cytotoxic property in this condition. LPS could inhibit the NOS activity (P<0.05), intracellular and extracellular NO production (P<0.05), proceeding of cell cycle (P<0.05); increase intracellular ROS production (P<0.01) and percentage of cell apoptosis (P<0.05). Attentively, TMP could dose-dependently inhibit all the effect mediated by LPS as described above.Conclusion TMP can attenuate the damage in HUVECs induced by LPS through inhibiting the decrease of NO production level and ROS increasing.