Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (20): 3655-3659.doi: 10.3969/j.issn.1673-8225.2010.20.011

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Transfection of cardiac microvascular endothelial cells using TrkB-shiRNA plasmid

Chen Si-yun1,2,3, Cao Liang1,2,3, Li Zhen1,2,3, Shen Xiao-tao1,2,3,4, Zheng Xin1,2,3, Liang Yong-jia1,2,3, Cai Dong-qing1,2,3,4   

  1. 1Key Laboratory for Regenerative Medicine, Ministry of Education, 2Joint Laboratory for Regenerative Medicine, the Chinese University of Hong KongJinan University, 3Guangdong Provincial Department of Science and Technology, International Science & Technology Cooperation Base, 4Department of Biomedical Engineering, Jinan University, Guangzhou  510632, Guangdong Province, China
  • Online:2010-05-14 Published:2010-05-14
  • Contact: Cai Dong-qing, Professor, Doctoral supervisor, Key Laboratory for Regenerative Medicine, Ministry of Education, Joint Laboratory for Regenerative Medicine, the Chinese University ofHong Kong-Jinan University, Guangdong Provincial Department of Science and Technology, International Science & Technology Cooperation Base, Department of Biomedical Engineering, Jinan University, Guangzhou 510632, Guangdong Province, China tdongbme@jnu.edu.cn
  • About author:Chen Si-yun, Studying for master’s degree, Key Laboratory for Regenerative Medicine, Ministry of Education, Joint Laboratory for Regenerative Medicine, the Chinese University of Hong KongJinan University, Guangdong Provincial Department of Science and Technology, International Science & Technology Cooperation Base, Jinan University, Guangzhou 510632, Guangdong Province, China csy.csy@163.com
  • Supported by:

    the 863 Program, No. 2007AA02Z105*;
    the National Natural Science Foundation of China, No. 30770886*, 30570369*, 30340038*, 30973158*;
    Guangdong Key Grant for Natural Science Foundation, No. 04105826*;
    Guangdong Grant for Science and Technology Development, No. 2004B30601007*

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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.

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