目的研究蛇床子素对高糖诱导原代培养人脐静脉血管内皮细胞(HUVEC)炎症的保护作用。方法将HUVEC细胞分为对照组、高糖组、对照+蛇床子素组和高糖+蛇床子素组。培养5 d后,采用硝酸还原酶法测定细胞上清液中一氧化氮(NO),荧光酶标仪测定细胞内活性氧(ROS),并通过real-time PCR和Western blot法检测趋化因子CCL5及其受体CCR5 mRNA和蛋白的表达。结果高糖明显降低对照组细胞上清液中NO水平同时升高ROS,蛇床子素能增加高糖组细胞上清液中NO水平同时降低高糖组细胞内ROS;real-time PCR和Western blot结果显示蛇床子素可以降低高糖组细胞CCL5及其受体CCR5 mRNA和蛋白的表达量。结论蛇床子素对高糖诱导的人脐静脉血管内皮细胞的损害具有保护作用是与其抗氧化应激及抗炎作用有关。
AIM To study the protective effects of osthole on the inflammation of primary cultured human um- bilical vein endothelial cells (HUVEC) induced by high glucose, which provided the theoretical basis for the study of new anti-diabetes agents. METHODS HUVECs were divided into four groups: control, high glucose, control + osthole, and high glucose + osthole group. After being cultured 5 d, the nitric oxide (NO) level in super- natant was detected by nitrate reductase; intracellular reactive oxygen species (ROS) was analyzed with fluores- cence probe; the expressions of CCL5 and CCR5 were measured by the real-time PCR and Western blot. RE- SULTS High glucose markedly decreased NO level in the supernatant in the control group but increased its ROS. Osthole elevated NO level in the supernatant when used with the high glucose, as well as lessened its ROS level. The results of real-time PCR and Western blot showed that osthole could weaken the expressions of CCL5 and CCR5 mRNA, which were induced by high glucose. CONCLUSION Osthole has protective effects on the injury of HUVEC induced by high glucose. Its working mechanism might be related to the prevention and control of diabetes angiopathies.