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ENGLISH ABSTRACT
JOURNAL ARTICLE
RESEARCH SUPPORT, NON-U.S. GOV'T
[tMfn2 gene transfer promotes vascular smooth-muscle cells apoptosis via activation of the mitochondrial cell death pathway].
Zhonghua Xin Xue Guan Bing za Zhi 2009 July
OBJECTIVE: To investigate the effect of tMfn2 gene transfer on promoting the apoptosis of vascular smooth-muscle cells (VSMCs).
METHODS: VSMCs were infected by adenovirus-mediated tMfn2 (Adv-tMfn2) or adenovirus-mediated Mfn2 (Adv-Mfn2). FACS analysis, cell death ELISA and TUNEL staining were used to investigate the role of tMfn2 and Adv-Mfn2 gene transfer on VSMCs apoptosis. Western blot was used to analyze the protein expression of p-Akt, Bcl-2, Bax and cleaved caspase-9.
RESULTS: FACS and ELISA results showed that tMfn2 was superior to Mfn2 in promoting VSMCs apoptosis and tMfn2 gene transfer induced VSMCs apoptosis in a time-dependent manner (P < 0.01). TUNEL staining evidenced that there were more positive-apoptotic VSMCs in tMfn2 group than that in Mfn2 group (P < 0.01). The protein expressions of phosphorylated Akt and Bcl-2 were significantly decreased, whereas Bax and cleaved caspase-9 protein expressions were significantly upregulated in tMfn2-transfected VSMCs.
CONCLUSIONS: tMfn2 transfer promoted apoptosis of VSMCs in a time dependent manner via the PI3K-Akt signaling pathway and mitochondrial apoptotic pathway.
METHODS: VSMCs were infected by adenovirus-mediated tMfn2 (Adv-tMfn2) or adenovirus-mediated Mfn2 (Adv-Mfn2). FACS analysis, cell death ELISA and TUNEL staining were used to investigate the role of tMfn2 and Adv-Mfn2 gene transfer on VSMCs apoptosis. Western blot was used to analyze the protein expression of p-Akt, Bcl-2, Bax and cleaved caspase-9.
RESULTS: FACS and ELISA results showed that tMfn2 was superior to Mfn2 in promoting VSMCs apoptosis and tMfn2 gene transfer induced VSMCs apoptosis in a time-dependent manner (P < 0.01). TUNEL staining evidenced that there were more positive-apoptotic VSMCs in tMfn2 group than that in Mfn2 group (P < 0.01). The protein expressions of phosphorylated Akt and Bcl-2 were significantly decreased, whereas Bax and cleaved caspase-9 protein expressions were significantly upregulated in tMfn2-transfected VSMCs.
CONCLUSIONS: tMfn2 transfer promoted apoptosis of VSMCs in a time dependent manner via the PI3K-Akt signaling pathway and mitochondrial apoptotic pathway.
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