NF-κB信号通路对糖尿病血管内皮的影响

Effect of nuclear factor-kappa B signaling pathway on vascular endothelium of diabetes

目的研究糖尿病血管内皮细胞NF-κB信号途径,从而为防治糖尿病血管并发症提供新的治疗策略。方法设计、合成针对NF-κB基因ShRNA的寡核苷酸序列;构建腺病毒载体质粒,转染293A细胞,进行病毒包装、扩增;腺病毒载体转导血管内皮细胞,通过RT-PCR和Western blot从mRNA以及蛋白质水平验证构建的病毒载体对血管内皮细胞NF-κB表达的抑制效应;用Western blot验证NF-κB干扰的血管内皮细胞中JNK(c-Jun氨基末端激酶)的变化。结果高浓度葡萄糖的条件下,内皮细胞NF-κB的表达量显著升高;干扰RNA有效干扰了正常血糖和高血糖环境下NF-κB及p-JNK的表达;p-JNK的表达随着NF-κB表达的下降而下降。结论高糖通过NF-κB及JNK的信号传导通路影响内皮细胞的凋亡。

Objective To provide a new treatment strategy for the prevention of diabetes mellitus,through investigating the NF-kappa B signaling pathway of diabetic vascular endothelial cells.Methods The NF-kappa B gene ShRNA oligonucleotide sequence was designed and synthesized.Adenovirus vector plasmid which were transfected to 293 Acells were constructed,then were packaged and amplifyed.The virus was transducted to vascular endothelial cells,then the expression of inhibition of NFkappa B was proved by RT-PCR and Western blot from the mRNA and protein level in vascular endothelial cells.By using Western blot,the changes of JNK with the NF-kappa B disturbance in vascular endothelial cells was tested.Results Under the condition of high concentration of glucose,the expression of NF-kappa B in endothelial cells was significantly elevated.RNAi effectively interfered the expression of NF-kappa B and p-JNK on the condition of the normal blood sugar and high blood sugar.The expression of p-JNK decreased with the expression of NF-kappa B.Conclusion High sugar can affect the apoptosis of endothelial cells through the signaling pathway of NF-kappa B and p-JNK.