中文摘要：

目的观察川芎嗪对波动高糖诱导人脐静脉内皮细胞损伤及蛋白激酶Cβ1（PKCβ1）表达的影响。方法分离、鉴定并培养人脐静脉内皮细胞,实验共分为7组：正常低糖（N）组、稳定高糖（W）组、波动高糖（B）组、稳定高糖＋LY333531（WL）组、波动高糖＋LY333531（BL）组、波动高糖＋川芎嗪（TMP）组和波动高糖＋二甲双胍（MH）组。8d后检测各组细胞凋亡率、肿瘤坏死因子α（TNF-α）、可溶性细胞间黏附分子1（sICAM-1）、晚期氧化蛋白产物（AOPP）和总抗氧化能力（T-AOC）水平及细胞PKCβ1蛋白表达情况。结果与N组相比,W组和B组HUVECs总凋亡率、TNF-α、sICAM-1、AOPP含量、PKCβ1蛋白表达均显著升高（P〈0.01）,T-AOC显著降低（P〈0.01）,且W组和B组间差异有统计学意义（P〈0.05或P〈0.01）。应用TMP和MH干预具有与LY333531阻断相似的效应,可显著降低细胞的总凋亡率、TNF-α、sICAM-1、AOPP含量及PKCβ1蛋白表达（P〈0.01）,同时可显著升高T-AOC水平（P〈0.01）。结论波动性高糖状态具有显著加重人脐静脉内皮细胞损伤的效应,且该效应与PKCβ1表达水平显著升高密切相关;川芎嗪对波动性高血糖状态下的血管内皮功能具有明显的保护作用,其作用机制与其显著抑制PKCβ1的过表达进而减轻氧化应激和炎症反应密切相关。

英文摘要：

Objective To investigate the effects of Tetramethylpyrazine（TMP）on human umbilical vein endothelial cells（HUVECs）injury induced by high blood glucose fluctuations and the PKCβ1expression.Methods HUVECs were incubated for 8 days in media containing different glucose concentrations：5.56 mmol/L（normal glucose,N）,25 mmol/L（constant high glucose,W）,or a daily alternating 5.56 or 25 mmol/L glucose（fluctuant high glucose,B）.LY333531（200 nmol/L）was pretreated on W group（WL）and B group（BL）,respectively.Meanwhile,B group were treated with tetramethylpyrazine hydrochloride（500μmol/L）and metformin hydrochloride（1 mmol/L）,respectively,which were called as TMP group and MH group.Cell apoptosis was measured by flow cytometry.The concentrations of tumor necrosis factor-alpha（TNF-α）,soluble intercellular adhesion molecule-1（sICAM-1）,AOPP and total antioxidant capacity（T-AOC）in the cell culture supernatant were measured by ELISA method.The expression of protein kinase C（PKC）β1in HUVECs was measured by Western blot.Results In comparison with N group,cell apoptosis rate（mainly late apoptosis）,levels of TNF-α,sICAM-1 and AOPP,the expression of PKCβ1in W group and B group were all significantly increased（P〈0.05 or P〈0.01）,whereas T-AOC was significantly decreased（P〈0.01）,all especially in B group（P〈0.05 or P〈0.01）.Pretreated with LY333531,cell apoptosis rate,levels of TNF-α,sICAM-1 and AOPP,as well as the expression of PKCβ1,were all significantly decreased（P〈0.05 or P〈0.01）,meanwhile T-AOC was significantly increased（P〈0.05 or P〈0.01）.TMP and MH manifested the similar protective effects as LY333531.Conclusion High blood glucose can induce HUVEC injury obviously,which is closely related to PKCβ1over-expression.Tetramethylpyrazine can protect HUVECs suffering fluctuant high glucose obviously,and the protective mechanism of which is closely related to its effects of relieving vessel stress,alleviating inflammatory reaction and in