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Critical role of insulin‑like growth factor binding protein‑5 in methamphetamine‑induced apoptosis in cardiomyocytes.

Leung KP, Qu YH, Qiao DF, Xie WB, Li DR, Xu JT, Wang HJ, Yue X - Mol Med Rep (2014)

Bottom Line: The cardiotoxic effects have yet not been clearly elucidated with respect to the apoptotic pathway.Silencing IGFBP5 with small interfering RNA significantly reduced apoptosis and suppressed the expression of caspase‑3 in NRVMs following treatment with MA.To the best of our knowledge, the present study provided the first evidence suggesting that IGFBP5 is a potential therapeutic target in MA‑induced apoptosis in vitro, providing a foundation for future in vivo studies.

View Article: PubMed Central - PubMed

Affiliation: Department of Forensic Medicine, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China.

ABSTRACT
Methamphetamine (MA) is a highly abused amphetamine‑like psychostimulant. At present, the mechanisms underlying MA‑induced cardiotoxicity are poorly understood. The cardiotoxic effects have yet not been clearly elucidated with respect to the apoptotic pathway. Insulin‑like growth factor binding protein‑5 (IGFBP5) is important for cell growth control and the induction of apoptosis. The aim of the present study was to analyze whether IGFBP5 is involved in MA‑induced apoptosis as a novel target. MA‑induced apoptosis was observed in neonatal rat ventricular myocytes (NRVMs) in a concentration‑dependent manner using a terminal deoxyribonucleotide transferase‑mediated dUTP nick end‑labeling assay. Using reverse transcription polymerase chain reaction and western blotting, MA was demonstrated to induce concentration‑dependent increases in the expression of IGFBP5. Silencing IGFBP5 with small interfering RNA significantly reduced apoptosis and suppressed the expression of caspase‑3 in NRVMs following treatment with MA. To the best of our knowledge, the present study provided the first evidence suggesting that IGFBP5 is a potential therapeutic target in MA‑induced apoptosis in vitro, providing a foundation for future in vivo studies.

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Related in: MedlinePlus

The mRNA and protein expression of IGFBP5 increases in a dose-dependent manner in NRVMs treated with MA for 48 h. NRVMs were incubated with MA at varying concentrations (0, 0.5, 1.0 or 1.5 mM). (A) Total RNA was subjected to reverse transcription polymerase chain reaction. (B) Whole cell lysates were analyzed by western blotting. An antibody against β-actin was used as the loading control. The data are presented as the mean ± standard deviation (n=6; *P<0.01). MA, methamphetamine; NRVMs, neonatal rat ventricular myocytes; IGFBP5, insulin-like growth factor binding protein-5.
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f2-mmr-10-05-2306: The mRNA and protein expression of IGFBP5 increases in a dose-dependent manner in NRVMs treated with MA for 48 h. NRVMs were incubated with MA at varying concentrations (0, 0.5, 1.0 or 1.5 mM). (A) Total RNA was subjected to reverse transcription polymerase chain reaction. (B) Whole cell lysates were analyzed by western blotting. An antibody against β-actin was used as the loading control. The data are presented as the mean ± standard deviation (n=6; *P<0.01). MA, methamphetamine; NRVMs, neonatal rat ventricular myocytes; IGFBP5, insulin-like growth factor binding protein-5.

Mentions: IGFBP5 mRNA was identified in NRVMs using qPCR (Fig. 2A). Melting curve analysis confirmed the specificity of transcripts of IGFBP5 in NRVMs. To evaluate how MA affects IGFBP5 expression, NRVMs were exposed to varying concentrations of MA. MA increased the mRNA expression of IGFBP5 in a dose-dependent manner. The mRNA expression of IGFBP5 significantly increased in the 1.5 mM MA group by 7.06±0.16 fold (n=6; P<0.01). IGFBP5 protein expression was also evaluated in MA-treated NRVMs (Fig. 2B). Immunoblotting using anti-IGFBP5 antibody indicated that the molecular weight of the IGFBP5 protein was 31 kDa. MA increased IGFBP5 protein expression in MA-treated NRVMs to 4.67±0.21 fold (n=6; P<0.01).


Critical role of insulin‑like growth factor binding protein‑5 in methamphetamine‑induced apoptosis in cardiomyocytes.

Leung KP, Qu YH, Qiao DF, Xie WB, Li DR, Xu JT, Wang HJ, Yue X - Mol Med Rep (2014)

The mRNA and protein expression of IGFBP5 increases in a dose-dependent manner in NRVMs treated with MA for 48 h. NRVMs were incubated with MA at varying concentrations (0, 0.5, 1.0 or 1.5 mM). (A) Total RNA was subjected to reverse transcription polymerase chain reaction. (B) Whole cell lysates were analyzed by western blotting. An antibody against β-actin was used as the loading control. The data are presented as the mean ± standard deviation (n=6; *P<0.01). MA, methamphetamine; NRVMs, neonatal rat ventricular myocytes; IGFBP5, insulin-like growth factor binding protein-5.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4214346&req=5

f2-mmr-10-05-2306: The mRNA and protein expression of IGFBP5 increases in a dose-dependent manner in NRVMs treated with MA for 48 h. NRVMs were incubated with MA at varying concentrations (0, 0.5, 1.0 or 1.5 mM). (A) Total RNA was subjected to reverse transcription polymerase chain reaction. (B) Whole cell lysates were analyzed by western blotting. An antibody against β-actin was used as the loading control. The data are presented as the mean ± standard deviation (n=6; *P<0.01). MA, methamphetamine; NRVMs, neonatal rat ventricular myocytes; IGFBP5, insulin-like growth factor binding protein-5.
Mentions: IGFBP5 mRNA was identified in NRVMs using qPCR (Fig. 2A). Melting curve analysis confirmed the specificity of transcripts of IGFBP5 in NRVMs. To evaluate how MA affects IGFBP5 expression, NRVMs were exposed to varying concentrations of MA. MA increased the mRNA expression of IGFBP5 in a dose-dependent manner. The mRNA expression of IGFBP5 significantly increased in the 1.5 mM MA group by 7.06±0.16 fold (n=6; P<0.01). IGFBP5 protein expression was also evaluated in MA-treated NRVMs (Fig. 2B). Immunoblotting using anti-IGFBP5 antibody indicated that the molecular weight of the IGFBP5 protein was 31 kDa. MA increased IGFBP5 protein expression in MA-treated NRVMs to 4.67±0.21 fold (n=6; P<0.01).

Bottom Line: The cardiotoxic effects have yet not been clearly elucidated with respect to the apoptotic pathway.Silencing IGFBP5 with small interfering RNA significantly reduced apoptosis and suppressed the expression of caspase‑3 in NRVMs following treatment with MA.To the best of our knowledge, the present study provided the first evidence suggesting that IGFBP5 is a potential therapeutic target in MA‑induced apoptosis in vitro, providing a foundation for future in vivo studies.

View Article: PubMed Central - PubMed

Affiliation: Department of Forensic Medicine, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China.

ABSTRACT
Methamphetamine (MA) is a highly abused amphetamine‑like psychostimulant. At present, the mechanisms underlying MA‑induced cardiotoxicity are poorly understood. The cardiotoxic effects have yet not been clearly elucidated with respect to the apoptotic pathway. Insulin‑like growth factor binding protein‑5 (IGFBP5) is important for cell growth control and the induction of apoptosis. The aim of the present study was to analyze whether IGFBP5 is involved in MA‑induced apoptosis as a novel target. MA‑induced apoptosis was observed in neonatal rat ventricular myocytes (NRVMs) in a concentration‑dependent manner using a terminal deoxyribonucleotide transferase‑mediated dUTP nick end‑labeling assay. Using reverse transcription polymerase chain reaction and western blotting, MA was demonstrated to induce concentration‑dependent increases in the expression of IGFBP5. Silencing IGFBP5 with small interfering RNA significantly reduced apoptosis and suppressed the expression of caspase‑3 in NRVMs following treatment with MA. To the best of our knowledge, the present study provided the first evidence suggesting that IGFBP5 is a potential therapeutic target in MA‑induced apoptosis in vitro, providing a foundation for future in vivo studies.

Show MeSH
Related in: MedlinePlus