高糖诱导血管内皮细胞表达结缔组织生长因子的影响

High glucose promotes connective tissue growth factor expression in vascular endothelial cells with TGFβ-dependent pathways

目的观察高糖对血管内皮细胞表达结缔组织生长因子(connective tissue growth facotr,CTGF)的影响。方法体外培养人脐静脉内皮细胞(human umibilical vein endotheial cells,HUVECs),分别在含正常葡萄糖(5.5 mM)、高糖(25 mM葡萄糖)、25 mM甘露醇及高糖加转化生长因子β1(transforming frowth factorβ1,TGFβ1)中和抗体的培养基中培养24、48或72小时,并在相应时间点收集细胞总RNA及总蛋白。分别用半定量RT-PCR及Western Blot方法检测CTGF mRNA及蛋白表达水平的变化。结果与正常葡萄糖培养及25mM甘露醇对照组相比,高糖刺激24小时后,HUVECs的CTGF mRNA及蛋白表达水平均显著增加,并且这种作用持续到48小时。TGFβ1的中和抗体能部分阻断高糖的这种刺激作用。结论高糖能诱导HUVECs表达CTGF,而内皮细胞CTGF的表达增加可能是高糖参与动脉粥样硬化等心血管疾病发生的机制之一。

Objective Vascular endothelial dysfunction is characterized by multiple interactions between endothelial cells and components of the blood. This study was designed to study the effect of high glucose on pro-atherogenic connective tissue growth factor （CTGF） in vascular endothelial cells. Methods Cultured human umbilical vein endothelial cells （HUVECs） were treated with normal glucose （5.5 mM）, high glucose （25 mM）, 25mM mannitol or high glucose to- gether with a neutralizing antibody to transforming growth factor β1（TGF-β1） for 24, 48 or 72 hours respectively. The expression of CTGF mRNA and protein was determined with RT-PCR and western Blot, respectively. Results Expression of CTGF mRNA was up-regulated 24 hour after exposed to high glucose condition, which remained the elevated level up to 48 hours. Meanwhile, CTGF protein level increased after high glucose exposure for 24 hours, and maintained elevated status up to 48 hours. The upregulation of CTGF appears to be TGFβ-dependent since an anti-TGFβ neutralizing antibody partially blocks the effect of high glucose on CTGF gene and protein expression. Conclusion High glucose-induced CTGF expression in vascular endothelial cells is mediated, at least partially, with the TGF-β-dependent pathways, which suggests that CTGF might be an important factor involved in the patho-physiological responses to high glucose in vascular endothelial cells.