Limits...
CXCR4 attenuates cardiomyocytes mitochondrial dysfunction to resist ischaemia-reperfusion injury.

Cai WF, Kang K, Huang W, Liang JL, Feng YL, Liu GS, Chang DH, Wen ZL, Paul C, Xu M, Millard RW, Wang Y - J. Cell. Mol. Med. (2015)

Bottom Line: The increased CXCR4 expression was observed in cardiomyocytes post CXCR4-adenovirus transduction and this OE significantly reduced the cardiomyocyte contractility under basal conditions.Interestingly, this CXCR4-induced mitochondrial protective effect is associated with the enhanced signal transducer and activator of transcription 3 (expression in mitochondria.Consequently, in the presence of H/R, mitochondrial dysfunction was mitigated and cardiomyocyte death was decreased to 65% in the CXCR4 OE group as compared with the control group.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology & Lab Medicine, College of Medicine, University of Cincinnati, Cincinnati, OH, USA.

No MeSH data available.


Related in: MedlinePlus

Hypoxia-reoxygenation-induced apoptosis is associated with the decreased CXCR4 expression in rat cardiomyocytes. (A) Representative immunoblots illustrating CXCR4, cleaved-caspase3 and GAPDH in response to the hypoxia/reoxygenation (H/R). (B and C) Quantitative analysis of CXCR4 (B) and cleaved-Caspase3 (C) expression levels relative to normal condition after normalization to GAPDH levels. (D) DNA fragmentation (mono- and oligo-nucleosomes contents) in isolated rat myocytes was detected by ELISA in presence and absence of H/R injury. (E) Representative flow cytometric pseudo-colour density plots illustrating the distribution of apoptotic (PE-Cy7 positive) and necrotic (eFluor-780 positive) cells. (F) Quantitative analysis of annexin V or eFluor-780 positive cells in either the absence or presence of HR injury. (G) ADP and ATP concentrations were assessed by colorimetric assay, and the ratios of ADP to ATP were used to determine the energetic metabolism of cardiomyocytes either in the absence or presence of H/R. n represents six independent experiments; *P < 0.05 versus normal group; #P < 0.05 versus H/R 1 and 4 hrs.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4549033&req=5

fig01: Hypoxia-reoxygenation-induced apoptosis is associated with the decreased CXCR4 expression in rat cardiomyocytes. (A) Representative immunoblots illustrating CXCR4, cleaved-caspase3 and GAPDH in response to the hypoxia/reoxygenation (H/R). (B and C) Quantitative analysis of CXCR4 (B) and cleaved-Caspase3 (C) expression levels relative to normal condition after normalization to GAPDH levels. (D) DNA fragmentation (mono- and oligo-nucleosomes contents) in isolated rat myocytes was detected by ELISA in presence and absence of H/R injury. (E) Representative flow cytometric pseudo-colour density plots illustrating the distribution of apoptotic (PE-Cy7 positive) and necrotic (eFluor-780 positive) cells. (F) Quantitative analysis of annexin V or eFluor-780 positive cells in either the absence or presence of HR injury. (G) ADP and ATP concentrations were assessed by colorimetric assay, and the ratios of ADP to ATP were used to determine the energetic metabolism of cardiomyocytes either in the absence or presence of H/R. n represents six independent experiments; *P < 0.05 versus normal group; #P < 0.05 versus H/R 1 and 4 hrs.

Mentions: Isolated rat cardiomyocytes were subjected to H/R condition to investigate the alteration of CXCR4 expression level in response to I/R injury. After exposure to 3 hrs of hypoxia and 1 hr of re-oxygenation, CXCR4 expression level was not changed significantly. However, the expression level was decreased by 36%, 55% and 77% after re-oxygenation for 4, 7 and 9 hrs, respectively, when compared to cardiomyocytes maintained under normal control conditions (Fig.1A and B). Immunoblot analysis indicated that the appearance of the cleaved form of caspase-3 was enhanced to 1.20-, 1.23-, 1.80- and 2.0-fold in response to 1, 4, 7 and 9 hrs of re-oxygenation compared to normal status (Fig.1C). During apoptosis, activated caspase-3 can cleave nuclear DNA between nucleosomes, producing a mixture of DNA fragments. Consistent with this, the ELISA assay showed that the DNA fragmentation increased in a time-dependent pattern post H/R (Fig.1D). Indeed, both apoptosis and necrosis were increased which demonstrated by the enhanced Annexin V and eFluor-780 positive cells respectively (Fig.1E and F). Cellular bioenergetics, as is indicated by the ADP/ATP ratio, also increased in a time-dependent manner after cardiomyocyte exposure to H/R injury (Fig.1G).


CXCR4 attenuates cardiomyocytes mitochondrial dysfunction to resist ischaemia-reperfusion injury.

Cai WF, Kang K, Huang W, Liang JL, Feng YL, Liu GS, Chang DH, Wen ZL, Paul C, Xu M, Millard RW, Wang Y - J. Cell. Mol. Med. (2015)

Hypoxia-reoxygenation-induced apoptosis is associated with the decreased CXCR4 expression in rat cardiomyocytes. (A) Representative immunoblots illustrating CXCR4, cleaved-caspase3 and GAPDH in response to the hypoxia/reoxygenation (H/R). (B and C) Quantitative analysis of CXCR4 (B) and cleaved-Caspase3 (C) expression levels relative to normal condition after normalization to GAPDH levels. (D) DNA fragmentation (mono- and oligo-nucleosomes contents) in isolated rat myocytes was detected by ELISA in presence and absence of H/R injury. (E) Representative flow cytometric pseudo-colour density plots illustrating the distribution of apoptotic (PE-Cy7 positive) and necrotic (eFluor-780 positive) cells. (F) Quantitative analysis of annexin V or eFluor-780 positive cells in either the absence or presence of HR injury. (G) ADP and ATP concentrations were assessed by colorimetric assay, and the ratios of ADP to ATP were used to determine the energetic metabolism of cardiomyocytes either in the absence or presence of H/R. n represents six independent experiments; *P < 0.05 versus normal group; #P < 0.05 versus H/R 1 and 4 hrs.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4549033&req=5

fig01: Hypoxia-reoxygenation-induced apoptosis is associated with the decreased CXCR4 expression in rat cardiomyocytes. (A) Representative immunoblots illustrating CXCR4, cleaved-caspase3 and GAPDH in response to the hypoxia/reoxygenation (H/R). (B and C) Quantitative analysis of CXCR4 (B) and cleaved-Caspase3 (C) expression levels relative to normal condition after normalization to GAPDH levels. (D) DNA fragmentation (mono- and oligo-nucleosomes contents) in isolated rat myocytes was detected by ELISA in presence and absence of H/R injury. (E) Representative flow cytometric pseudo-colour density plots illustrating the distribution of apoptotic (PE-Cy7 positive) and necrotic (eFluor-780 positive) cells. (F) Quantitative analysis of annexin V or eFluor-780 positive cells in either the absence or presence of HR injury. (G) ADP and ATP concentrations were assessed by colorimetric assay, and the ratios of ADP to ATP were used to determine the energetic metabolism of cardiomyocytes either in the absence or presence of H/R. n represents six independent experiments; *P < 0.05 versus normal group; #P < 0.05 versus H/R 1 and 4 hrs.
Mentions: Isolated rat cardiomyocytes were subjected to H/R condition to investigate the alteration of CXCR4 expression level in response to I/R injury. After exposure to 3 hrs of hypoxia and 1 hr of re-oxygenation, CXCR4 expression level was not changed significantly. However, the expression level was decreased by 36%, 55% and 77% after re-oxygenation for 4, 7 and 9 hrs, respectively, when compared to cardiomyocytes maintained under normal control conditions (Fig.1A and B). Immunoblot analysis indicated that the appearance of the cleaved form of caspase-3 was enhanced to 1.20-, 1.23-, 1.80- and 2.0-fold in response to 1, 4, 7 and 9 hrs of re-oxygenation compared to normal status (Fig.1C). During apoptosis, activated caspase-3 can cleave nuclear DNA between nucleosomes, producing a mixture of DNA fragments. Consistent with this, the ELISA assay showed that the DNA fragmentation increased in a time-dependent pattern post H/R (Fig.1D). Indeed, both apoptosis and necrosis were increased which demonstrated by the enhanced Annexin V and eFluor-780 positive cells respectively (Fig.1E and F). Cellular bioenergetics, as is indicated by the ADP/ATP ratio, also increased in a time-dependent manner after cardiomyocyte exposure to H/R injury (Fig.1G).

Bottom Line: The increased CXCR4 expression was observed in cardiomyocytes post CXCR4-adenovirus transduction and this OE significantly reduced the cardiomyocyte contractility under basal conditions.Interestingly, this CXCR4-induced mitochondrial protective effect is associated with the enhanced signal transducer and activator of transcription 3 (expression in mitochondria.Consequently, in the presence of H/R, mitochondrial dysfunction was mitigated and cardiomyocyte death was decreased to 65% in the CXCR4 OE group as compared with the control group.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology & Lab Medicine, College of Medicine, University of Cincinnati, Cincinnati, OH, USA.

No MeSH data available.


Related in: MedlinePlus