【摘要】 背景:脑源性神经营养因子与其受体TrkB在心脏与骨骼肌内皮细胞存在表达,在心脏血管系统的发育中及骨骼肌缺血时能有效促进血管新生,但其促血管新生的细胞与分子机制尚不清楚。目的:应用针对脑源性神经营养因子受体TrkB的shiRNA质粒对心脏微血管内皮细胞进行转染,观察TrkB 3种亚型的表达及心脏微血管内皮细胞的生长状况和形态,初步探讨脑源性神经营养因子-TrkB通路在心脏微血管内皮细胞中的调控作用。方法:使用TrkB-shiRNA质粒转染大鼠心脏微血管内皮细胞,应用荧光实时定量PCR检测TrkB的3个亚型,TrkB-FL、TrkB-T1及TrkB-T2 mRNA的表达,并观察经转染的心脏微血管内皮细胞的增殖情况。结果与结论:应用TrkB-shiRNA质粒转染心脏微血管内皮细胞后,TrkB 3种亚型在转染后的4d内mRNA表达均下降(P<0.05或P<0.01),且经转染的心脏微血管内皮细胞的生长变慢。说明TrkB-shiRNA质粒可沉默心脏微血管内皮细胞的TrkB-FL、TrkB-T1及TrkB-T2的表达,且TrkB表达被抑制可能会影响心脏微血管内皮细胞的增殖。更多还原

【Abstract】 BACKGROUND: Recent studies have shown that endothelial cells (ECs) in heart and skeletal muscle express brain derived neurotrophic factor (BDNF) and its receptor tyrosine receptor kinase B (TrkB). BDNF-TrkB pathway might play an important role in cardiovascular system development, angiogenesis and ischemic limb regeneration. However, the molecular mechanism regarding to BDNF-induced angiogenesis is still unknown. OBJECTIVE: TrkB-shiRNA vector was transfected into cardiac microvascular endothelial cells (CMECs) to investigate the expression of 3 TrkB isoforms, the growth and cell morphology, in addition, to explore the regulation of TrkB pathway on CMECs. METHODS: TrkB-shiRNA vector was used. The expressions of TrkB isoforms (TrkB-FL, TrkB-T1 and TrkB-T2) in CMECs was analyzed by real-time PCR. The silencing effect of TrkBs in CMECs proliferation was analyzed by cell culture and cell counting. RESULTS AND CONCLUSION: It was found that the expression of TrkB isoforms, TrkB-FL, TrkB-T1 and TrkB-T2, was decreased at 4 days after CMECs transfected with TrkB-shiRNA vector (P < 0.05 or P < 0.01). In addition, the proliferation of transfected CMECs was decreased. It demonstrated that transfection of CMECs with TrkB-shiRNA vector was able to decrease expression of TrkB-FL, TrkB-T1 and TrkB-T2 and decrease the proliferation of CMECs.更多还原