不同浓度葡萄糖对血管内皮细胞11β-羟基类固醇脱氢酶基因表达的影响

The expression of 11β-hydroxysteroid dehydrogenase in diabetes mellitus vasculopathy treated with different concentration of glucose

目的:观察葡萄糖下对大动脉内皮细胞11β-HSD1,11β-HSD2表达的影响,及11β-HSD1与TGF-β1,ET-1等细胞因子之间的关系;同时观察反义寡核苷酸对高糖条件下大动脉内皮细胞11β-HSD1,11β-HSD2表达的干预作用。方法:猪髂动脉内皮细胞分别培养于不同浓度葡萄糖(5.6,10.0,20.0,30.0mmol/L)中,用RT-PCR法检测11β-HSD1,11β-HSD2,TGF-β1,ET-1 mRNA的表达,Western-blot方法检测11β-HSD1,11β-HSD2蛋白表达情况。结果:高糖状态干预48h后,11β-HSD1,TGF-β1 mRNA表达随葡萄糖浓度的增加而增加,呈现一定的相关性;11β-HSD1蛋白表达随葡萄糖浓度增加而增加。采用11β-HSD1与11β-HSD2反义寡核苷酸干预后11β-HSD1与11β-HSD2 mRNA和蛋白质表达完全被抑制,同时TGF-β1的表达降低。结论:葡萄糖刺激血管内皮细胞11β-HSD1 mRNA和蛋白质表达增加,且在高糖状态下,11β-HSD1 mRNA的表达与TGF-β1RNA表达有直线相关性,11β-HSD1可能与糖尿病及其血管并发症相关。

Objective：To investigate the effect of glucose and glucocorticoid on the expression of 11β-HSD1,11β-HSD2 gene transcriptions in PIEC in vitro,and the relationship between 11β-HSD1 and transforming growth factor-β1 and Endothelin-1. Meanwhile,to observe the effect of 11β-HSD1 and 11β-HSD2 antisense oligonucleotides on the expression of 11β-HSD1 and 11β-HSD2 by PIEC under hyperglycemia, nethods：PIEC were cultured in medium in vitro under different concentration of glucose （5.6 mmol/ L, 10.0 mmol/L,20.0 mmol/L,30.0 mmol/L）and in different concentration of glucocorticoid and antisense oligonucleotides under hyperglycemia. RT-PCR was applied to determine the expression of 11β-HSD1,11β-HSD2,TGF-β1 and ET-1 mRNA. Western-blot was performed to measurer the expression of 11β-HSD1 and 11β-HSD2. Results：After treatment of different concentration of glucose for 48 h,the expression of 1 1β-HSD1 and TGF-β1 mRNA under high glucose condition were significantly increased and represented a certain correlation;high glucose can increase the expression of 11β-HSD1 protein. Both mRNA and protein of 11β-HSD1 and 11β-HSD2 were blocked by 11β-HSD1 and 11β-HSD2 antisense oligonucleotides; meanwhile,the expression of TGF-β1 mRNA was decreased. Conclusion：High glucose and glucocorticoid could increase the expression of 11β-HSD1 mRNA and protein,there was a certain linear correlation between the expression of 11β-HSD 1 and TGF-β1 mRNA. 11β-HSD1 might have a correlation with diabetes mellitus.