摘要
目的观察高糖作用人脐静脉内皮细胞时血管内皮生长因子mRNA和蛋白的表达与蛋白激酶C通路被激活和抑制的关系。方法提取并体外培养人脐静脉内皮细胞,分别加入不同浓度葡萄糖(5.5、11、22及44mmol/L)、蛋白激酶C激活剂和阻断剂并分别作用不同时间,应用逆转录聚合酶链反应检测血管内皮生长因子mR-NA水平,免疫细胞化学检测血管内皮生长因子及蛋白激酶C蛋白表达。结果经高糖处理的人脐静脉内皮细胞中血管内皮生长因子mRNA的转录水平明显高于对照组(P<0.05或P<0.01),但脐静脉内皮细胞经22 mmol/L葡萄糖作用72 h或佛波酯作用6 h,血管内皮生长因子mRNA水平不再升高,而呈下降趋势(P<0.05)。经22 mmol/L葡萄糖作用72 h后,血管内皮生长因子蛋白表达也逐渐下降。蛋白激酶C在22 mmol/L葡萄糖处理2 h后出现膜转位。而蛋白激酶C抑制剂GF109203X可阻断上述现象。结论高糖能引起脐静脉内皮细胞血管内皮生长因子mR-NA和蛋白表达增高。高糖引起脐静脉内皮细胞血管内皮生长因子基因表达增高,其机制与高糖激活蛋白激酶C通路有关。
Aim To observe the transcript and protein expression levels of vascular endothelial growth factor （VEGF） in human umbilical vein endothelial cells （hUVEC） cultured with high glucose in vitro and investigate the relationship between it and protein kinase C （PKC） pathway. Methods hUVEC were cultured in vitro with glucose at different concentrations or for different cultural time and cultured with PKC inhibitor GF109203X or PKC activator PMA respectively. The transcript of VEGF was measured by RT-PCR. The protein expression of VEGF and PKCβB Ⅱ was detected with immtmocytochemistry. Results The transcript and protein expression levels of VFGF in high glucose groups or PKC activator PMA group were higher than control group or PKC inhibitor GF109203X group （ P〈0.05 or P〈0.01 ）. Cultured with glucose at 22 mmol/L, the mRNA and protein expression of VEGF increased in 72 h, the protein expression of VEGF reached the summit at 72 h, and then began to decrease （ P〈0.01 ）. Cultured with PMA, the transcript of VEGF increased within 6 h, and then decreased. The protein of PKCβⅡ .seemed to translocate to cell membrane from nucleus after treated with 22 mmol/L glucose for 2 h. However, PKC inhibitor GF109203X can inhibit the phenomena above. Conclusions The transcript and protein expression of VEGF gene increased in hUVEC, cultured with high glucose. High glucose induced the expression of VEGF gene via PKC pathway in h UVEC.
出处
《中国动脉硬化杂志》
CAS
CSCD
2006年第1期17-20,共4页
Chinese Journal of Arteriosclerosis
基金
湖南省教委基金
卫生厅科研基金(97B103)资助
