BDNF参与保护葡萄糖饥饿引起的微血管内皮细胞凋亡

BDNF involved in the protection of microvascular endothelial cell apoptosis under glucose starvation

目的:探讨脑源性神经营养因子(BDNF)是否参与葡萄糖缺乏状态下微血管内皮细胞的保护。方法:使用小鼠脑微血管内皮细胞bEnd.3细胞株构建葡萄糖饥饿模型,分别使用BDNF的siRNA及BDNF-TrkB通路阻断剂对经建模的微血管内皮细胞进行处理,使用流式细胞技术检测细胞凋亡,使用Western blot和qRT-PCR分别对BDNF及其他调控基因的表达进行研究。结果:葡萄糖饥饿会导致微血管内皮细胞凋亡,晚期凋亡水平随着缺糖时间增加而升高,且微血管内皮细胞BDNF表达量上调。在葡萄糖饥饿状态下,使用BDNF受体TrkB的抑制剂K252a阻断BDNF-TrkB通路能增加微血管内皮细胞的早期凋亡;抑制BDNF表达使微血管内皮细胞的凋亡升高。结论:BDNF参与保护葡萄糖饥饿引起的微血管内皮细胞凋亡,并参与非葡萄糖饥饿和葡萄糖饥饿状态下糖酵解关键调控基因PFKM表达水平的维持,从而在维持血管内皮细胞的存活及通过糖酵解供能中发挥重要的调控作用。

Objective:To investigate whether brain-derived neurotrophic factor(BDNF)is involved in protection of the survival of microvascular endothelial cells(MECs)under glucose starvation.Methods:The glucose starvation model was established using mouse brain microvascular endothelial cell line,bEnd.3.The modeled MECs were treated with siRNA of BDNF and BDNF-TrkB pathway blockers respectively.The flow cytometry was applied to investigate the apoptosis of treated cells.Western blot and qRT-PCR were used to analyze the expression of BDNF and other regulatory genes.Results:Glucose starvation leaded to apoptosis of MECs,and the degree of late apoptosis was increased with the prolongation of glucose starvation.With glucose starvation,the expression of BDNF in MECs was upregulated.Under glucose starvation,when BDNF-TrkB pathway was blocked with inhibitor of BDNF receptor,K252a,the early apoptosis of MECs was increased.In addition,inhibition of BDNF expression was able to increase apoptosis of MECs in glucose starvation.Conclusion:BDNF is involved in protection of the apoptosis of MECs mediated by glucose starvation,and in the maintenance of the expression of PFKM gene,a key regulatory gene of glycolysis,in the non-glucose starvation and glucose starvation.Thus plays an important regulatory role in maintaining the survival of MECs and energy supporting of MECs viaglycolysis.