Activation of IL-11/STAT3 pathway in preconditioned human skeletal myoblasts blocks apoptotic cascade under oxidant stress

Niagara Muhammad Idris, Muhammad Ashraf, Rafeeq P.H. Ahmed, Jiang Shujia, Khawaja H. Haider

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Aim: To determine whether our novel approach of diazoxide-induced stem cell preconditioning might be extrapolated to human skeletal myoblasts to support their survival under lethal oxidant stress. Methods & results: Using an in vitro model of H 2O 2 treatment of human skeletal myoblasts, we report the ability of diazoxide-preconditioned human skeletal myoblasts to express cytokines and growth factors, which act in an autocrine and paracrine fashion to promote their own survival. Preconditioning of skeletal myoblasts was cytoprotective and significantly reduced their apoptotic index (p < 0.05). IL-11 gene and protein expression was significantly increased in preconditioned skeletal myoblasts. Transfection of skeletal myoblasts with IL-11-specific siRNA incurred their death under oxidant stress. The cytoprotective effect of diazoxide preconditioning was blocked by Erk1/2 inhibitor PD98059 (20-100 μM), which abrogated STAT-3 phosphorylation, thus confirming a possible involvement of Erk1/2/STAT3 signaling downstream of IL-11 in cell survival. We also investigated the time course of subcellular changes and signaling pathway of skeletal myoblasts apoptosis under oxidant stress before and after preconditioning. Apoptosis was induced in skeletal myoblasts with 100-500 μM H 2O 2 for time points ranging from 1 to 24 h. Release of lactate dehydrogenase, disruption of the mitochondrial membrane potential and cytochrome-c translocation into cytoplasm were the earliest signs of apoptosis. Total Akt protein remained unchanged whereas marked reduction in pAkt was observed in the native skeletal myoblasts. Terminal dUTP nick end-labeling and annexin-V positivity were significantly increased after 4 h. Ultra-structure studies showed condensed chromatin, shriveled nuclei and swollen mitochondria. Conclusion: These data suggest that skeletal myoblasts undergo apoptosis under oxidant stress in a time-dependent manner and preconditioning of skeletal myoblasts significantly prevented their apoptosis via IL-11/STAT3 signaling.

Original languageEnglish (US)
Pages (from-to)47-57
Number of pages11
JournalRegenerative Medicine
Volume7
Issue number1
DOIs
StatePublished - Jan 1 2012
Externally publishedYes

Fingerprint

Skeletal Myoblasts
Interleukin-11
Cell death
Oxidants
Chemical activation
Proteins
Diazoxide
Apoptosis
Phosphorylation
Mitochondria
Stem cells
Labeling
Genes
Cells
Membranes
Survival
Mitochondrial Membrane Potential
Annexin A5
Cytochromes c
L-Lactate Dehydrogenase

Keywords

  • apoptosis
  • diazoxide
  • myoblasts
  • preconditioning

ASJC Scopus subject areas

  • Biomedical Engineering
  • Embryology

Cite this

Activation of IL-11/STAT3 pathway in preconditioned human skeletal myoblasts blocks apoptotic cascade under oxidant stress. / Idris, Niagara Muhammad; Ashraf, Muhammad; Ahmed, Rafeeq P.H.; Shujia, Jiang; Haider, Khawaja H.

In: Regenerative Medicine, Vol. 7, No. 1, 01.01.2012, p. 47-57.

Research output: Contribution to journalArticle

Idris, Niagara Muhammad ; Ashraf, Muhammad ; Ahmed, Rafeeq P.H. ; Shujia, Jiang ; Haider, Khawaja H. / Activation of IL-11/STAT3 pathway in preconditioned human skeletal myoblasts blocks apoptotic cascade under oxidant stress. In: Regenerative Medicine. 2012 ; Vol. 7, No. 1. pp. 47-57.
@article{5233157987a84f958d5ac3219c6113be,
title = "Activation of IL-11/STAT3 pathway in preconditioned human skeletal myoblasts blocks apoptotic cascade under oxidant stress",
abstract = "Aim: To determine whether our novel approach of diazoxide-induced stem cell preconditioning might be extrapolated to human skeletal myoblasts to support their survival under lethal oxidant stress. Methods & results: Using an in vitro model of H 2O 2 treatment of human skeletal myoblasts, we report the ability of diazoxide-preconditioned human skeletal myoblasts to express cytokines and growth factors, which act in an autocrine and paracrine fashion to promote their own survival. Preconditioning of skeletal myoblasts was cytoprotective and significantly reduced their apoptotic index (p < 0.05). IL-11 gene and protein expression was significantly increased in preconditioned skeletal myoblasts. Transfection of skeletal myoblasts with IL-11-specific siRNA incurred their death under oxidant stress. The cytoprotective effect of diazoxide preconditioning was blocked by Erk1/2 inhibitor PD98059 (20-100 μM), which abrogated STAT-3 phosphorylation, thus confirming a possible involvement of Erk1/2/STAT3 signaling downstream of IL-11 in cell survival. We also investigated the time course of subcellular changes and signaling pathway of skeletal myoblasts apoptosis under oxidant stress before and after preconditioning. Apoptosis was induced in skeletal myoblasts with 100-500 μM H 2O 2 for time points ranging from 1 to 24 h. Release of lactate dehydrogenase, disruption of the mitochondrial membrane potential and cytochrome-c translocation into cytoplasm were the earliest signs of apoptosis. Total Akt protein remained unchanged whereas marked reduction in pAkt was observed in the native skeletal myoblasts. Terminal dUTP nick end-labeling and annexin-V positivity were significantly increased after 4 h. Ultra-structure studies showed condensed chromatin, shriveled nuclei and swollen mitochondria. Conclusion: These data suggest that skeletal myoblasts undergo apoptosis under oxidant stress in a time-dependent manner and preconditioning of skeletal myoblasts significantly prevented their apoptosis via IL-11/STAT3 signaling.",
keywords = "apoptosis, diazoxide, myoblasts, preconditioning",
author = "Idris, {Niagara Muhammad} and Muhammad Ashraf and Ahmed, {Rafeeq P.H.} and Jiang Shujia and Haider, {Khawaja H.}",
year = "2012",
month = "1",
day = "1",
doi = "10.2217/rme.11.109",
language = "English (US)",
volume = "7",
pages = "47--57",
journal = "Regenerative Medicine",
issn = "1746-0751",
publisher = "Future Medicine Ltd.",
number = "1",

}

TY - JOUR

T1 - Activation of IL-11/STAT3 pathway in preconditioned human skeletal myoblasts blocks apoptotic cascade under oxidant stress

AU - Idris, Niagara Muhammad

AU - Ashraf, Muhammad

AU - Ahmed, Rafeeq P.H.

AU - Shujia, Jiang

AU - Haider, Khawaja H.

PY - 2012/1/1

Y1 - 2012/1/1

N2 - Aim: To determine whether our novel approach of diazoxide-induced stem cell preconditioning might be extrapolated to human skeletal myoblasts to support their survival under lethal oxidant stress. Methods & results: Using an in vitro model of H 2O 2 treatment of human skeletal myoblasts, we report the ability of diazoxide-preconditioned human skeletal myoblasts to express cytokines and growth factors, which act in an autocrine and paracrine fashion to promote their own survival. Preconditioning of skeletal myoblasts was cytoprotective and significantly reduced their apoptotic index (p < 0.05). IL-11 gene and protein expression was significantly increased in preconditioned skeletal myoblasts. Transfection of skeletal myoblasts with IL-11-specific siRNA incurred their death under oxidant stress. The cytoprotective effect of diazoxide preconditioning was blocked by Erk1/2 inhibitor PD98059 (20-100 μM), which abrogated STAT-3 phosphorylation, thus confirming a possible involvement of Erk1/2/STAT3 signaling downstream of IL-11 in cell survival. We also investigated the time course of subcellular changes and signaling pathway of skeletal myoblasts apoptosis under oxidant stress before and after preconditioning. Apoptosis was induced in skeletal myoblasts with 100-500 μM H 2O 2 for time points ranging from 1 to 24 h. Release of lactate dehydrogenase, disruption of the mitochondrial membrane potential and cytochrome-c translocation into cytoplasm were the earliest signs of apoptosis. Total Akt protein remained unchanged whereas marked reduction in pAkt was observed in the native skeletal myoblasts. Terminal dUTP nick end-labeling and annexin-V positivity were significantly increased after 4 h. Ultra-structure studies showed condensed chromatin, shriveled nuclei and swollen mitochondria. Conclusion: These data suggest that skeletal myoblasts undergo apoptosis under oxidant stress in a time-dependent manner and preconditioning of skeletal myoblasts significantly prevented their apoptosis via IL-11/STAT3 signaling.

AB - Aim: To determine whether our novel approach of diazoxide-induced stem cell preconditioning might be extrapolated to human skeletal myoblasts to support their survival under lethal oxidant stress. Methods & results: Using an in vitro model of H 2O 2 treatment of human skeletal myoblasts, we report the ability of diazoxide-preconditioned human skeletal myoblasts to express cytokines and growth factors, which act in an autocrine and paracrine fashion to promote their own survival. Preconditioning of skeletal myoblasts was cytoprotective and significantly reduced their apoptotic index (p < 0.05). IL-11 gene and protein expression was significantly increased in preconditioned skeletal myoblasts. Transfection of skeletal myoblasts with IL-11-specific siRNA incurred their death under oxidant stress. The cytoprotective effect of diazoxide preconditioning was blocked by Erk1/2 inhibitor PD98059 (20-100 μM), which abrogated STAT-3 phosphorylation, thus confirming a possible involvement of Erk1/2/STAT3 signaling downstream of IL-11 in cell survival. We also investigated the time course of subcellular changes and signaling pathway of skeletal myoblasts apoptosis under oxidant stress before and after preconditioning. Apoptosis was induced in skeletal myoblasts with 100-500 μM H 2O 2 for time points ranging from 1 to 24 h. Release of lactate dehydrogenase, disruption of the mitochondrial membrane potential and cytochrome-c translocation into cytoplasm were the earliest signs of apoptosis. Total Akt protein remained unchanged whereas marked reduction in pAkt was observed in the native skeletal myoblasts. Terminal dUTP nick end-labeling and annexin-V positivity were significantly increased after 4 h. Ultra-structure studies showed condensed chromatin, shriveled nuclei and swollen mitochondria. Conclusion: These data suggest that skeletal myoblasts undergo apoptosis under oxidant stress in a time-dependent manner and preconditioning of skeletal myoblasts significantly prevented their apoptosis via IL-11/STAT3 signaling.

KW - apoptosis

KW - diazoxide

KW - myoblasts

KW - preconditioning

UR - http://www.scopus.com/inward/record.url?scp=83655182222&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=83655182222&partnerID=8YFLogxK

U2 - 10.2217/rme.11.109

DO - 10.2217/rme.11.109

M3 - Article

C2 - 22168497

AN - SCOPUS:83655182222

VL - 7

SP - 47

EP - 57

JO - Regenerative Medicine

JF - Regenerative Medicine

SN - 1746-0751

IS - 1

ER -