Insulin-like growth factor-1 protecting cardiomyocytes from apoptosis by down-regulating transcription factor basic transcription element binding protein through extracellular regulated kinase 1/2 pathway

doi:10.16098/j.issn.0529-1356.2015.03.007

Acta Anatomica Sinica ›› 2015 ›› Issue (3) : 329-335. DOI: 10.16098/j.issn.0529-1356.2015.03.007

ZHANG Jian-kai¹CUI Xiao-jun¹XU Xiao-ling²DING Bi-lan² LI Jin-ju²LI Tao³ WU Zhu-guo ^4*

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Abstract

Objective To investigate gene regulation mechanism of insulin-like growth factor-1 (IGF-1) anti-apoptotic effect on rat cardiomyocytes. Methods Primary neonatal rat cardiomyocytes (NRCMs) were cultured in vitro，IGF-1(10nmol/L) was added with different signal transduction pathway inhibitors ［phosphatidylinositol 3-kinase(PI3K), extracellular regulated kinase (ERK)1/2 and Raf-1］ respectively (20μmol/L). The gene expression of basic transcription element binding protein (BTEB) was detected by RT-PCR and Western blotting, by which the pathway of IGF-1 down-regulated BTEB gene expression was judged. NRCMs were treated with 100umol/L hydrogen peroxide (H2O2) to induce apoptosis. BTEB specific siRNA was transfected into the cells by Lipofectamine 2000.Myocardial cells apoptosis was detected by DNA-ladder analysis, Annexin V-FITC／PI dual staining，Caspase-3 activity assay and Hoechst33258 staining. Results The mRNA and protein expression levels of BTEB gene in NRCMs were down-regulated significantly after IGF-1 had stimulated for 60 minutes. Compared with control groups, BTEB mRNA and protein expression in ERK1/2 pathway inhibitor PD98059 group was significantly higher (P＜0.01).The apoptosis of NRCMs was induced by H₂O₂. Artificially inhibited BTEB gene expression with BTEB specific siRNA，BTEB mRNA and protein expression decreased obviously (P＜0.05). Compared with control group, the apoptotic rates of NRCMs induced by H₂O₂in IGF-1 group and BTEB specific siRNA groups were declined (all P＜0.05)，decreased Caspase-3 activity(all P＜0.05), attenuated DNA fragmentation and reduced apoptotic bodies were also observed in these groups. The anti-apoptotic effect of BTEB gene silencing on NRCMs was similar with that of IGF-1 treatment. Conclusion IGF-1 protects cardiomyocytes from apoptosis by down-regulating transcription factor BTEB through ERK1/2 pathway.

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ZHANG Jian-kai CUI Xiao-jun XU Xiao-ling DING Bi-lan LI Jin-ju LI Tao WU Zhu-guo*. Insulin-like growth factor-1 protecting cardiomyocytes from apoptosis by down-regulating transcription factor basic transcription element binding protein through extracellular regulated kinase 1/2 pathway[J]. Acta Anatomica Sinica. 2015(3): 329-335 https://doi.org/10.16098/j.issn.0529-1356.2015.03.007

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