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Catestatin attenuates endoplasmic reticulum induced cell apoptosis by activation type 2 muscarinic acetylcholine receptor in cardiac ischemia/reperfusion.

Liao F, Zheng Y, Cai J, Fan J, Wang J, Yang J, Cui Q, Xu G, Tang C, Geng B - Sci Rep (2015)

Bottom Line: Cardiomyocytes endogenously produced CST and its expression was reduced after I/R.CST pretreatment decreased apoptosis especially endoplasmic reticulum (ER) stress response during I/R.The protective effects of CST were blocked by extracellular signal-regulated kinases 1/2 (ERK1/2) inhibitor PD90895 and phosphoinositide 3-kinase (PI3 K) inhibitor wortmannin.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology and Pathophysiology, School of Basic Medical Science, Peking University, P.R. China.

ABSTRACT
Catestatin (CST) is a catecholamine secretion inhibiting peptide as non-competitive inhibitor of nicotinic acetylcholine receptor. CST play a protective role in cardiac ischemia/reperfusion (I/R) but the molecular mechanism remains unclear. Cardiomyocytes endogenously produced CST and its expression was reduced after I/R. CST pretreatment decreased apoptosis especially endoplasmic reticulum (ER) stress response during I/R. The protection of CST was confirmed in H9c2 cardiomyoblasts under Anoxia/reoxygenation (A/R). In contrast, siRNA-mediated knockdown of CST exaggerated ER stress induced apoptosis. The protective effects of CST were blocked by extracellular signal-regulated kinases 1/2 (ERK1/2) inhibitor PD90895 and phosphoinositide 3-kinase (PI3 K) inhibitor wortmannin. CST also increased ERK1/2 and protein kinase B (Akt) phosphorylation and which was blocked by atropine and selective type 2 muscarinic acetylcholine (M2) receptor, but not type 1 muscarinic acetylcholine (M1) receptor antagonist. Receptor binding assay revealed that CST competitively bound to the M2 receptor with a 50% inhibitory concentration of 25.7 nM. Accordingly, CST inhibited cellular cAMP stimulated by isoproterenol or forskolin, and which was blocked by selective M2 receptor antagonist. Our findings revealed that CST binds to M2 receptor, then activates ERK1/2 and PI3 K/Akt pathway to inhibit ER stress-induced cell apoptosis resulting in attenuation cardiac I/R injury.

No MeSH data available.


Related in: MedlinePlus

CST pretreatment reduced ER-stress and decreased apoptosis.(a) Cleaved caspase-3 immunofluorescence staining and nuclear staining; apoptotic cells were stained in pink Scale bar = 100 μM. (b) Apoptosis cell numbers were counted in five different field of vision. (c) Western blot analysis of effect of I/R and CP on apoptosis and caspase family members and PARP and (d) proteins related to the ER stress pathway. Relative protein expression was analyzed by band gray degree. β-actin is a normalization control (e). Six independent experiments were performed for above studies.
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f2: CST pretreatment reduced ER-stress and decreased apoptosis.(a) Cleaved caspase-3 immunofluorescence staining and nuclear staining; apoptotic cells were stained in pink Scale bar = 100 μM. (b) Apoptosis cell numbers were counted in five different field of vision. (c) Western blot analysis of effect of I/R and CP on apoptosis and caspase family members and PARP and (d) proteins related to the ER stress pathway. Relative protein expression was analyzed by band gray degree. β-actin is a normalization control (e). Six independent experiments were performed for above studies.

Mentions: We used cleaved caspase-3 (a marker of cell apoptosis) antibody immunofluorescence staining to assess cell apoptosis. I/R increased the number of apoptotic cardiomyocytes, and CP significantly reduced the cell apoptosis (Fig. 2a,b). Consistently, I/R increased apoptosis markers (caspase-9, 7 and -3 cleavage, and poly (ADP-ribose) polymerase (PARP) cleaved into 89- and 24-kDa segments). CP lowered cleaved caspase-9, -7, -3 and PARP (Fig. 2c,e) for a role in reduced apoptosis.


Catestatin attenuates endoplasmic reticulum induced cell apoptosis by activation type 2 muscarinic acetylcholine receptor in cardiac ischemia/reperfusion.

Liao F, Zheng Y, Cai J, Fan J, Wang J, Yang J, Cui Q, Xu G, Tang C, Geng B - Sci Rep (2015)

CST pretreatment reduced ER-stress and decreased apoptosis.(a) Cleaved caspase-3 immunofluorescence staining and nuclear staining; apoptotic cells were stained in pink Scale bar = 100 μM. (b) Apoptosis cell numbers were counted in five different field of vision. (c) Western blot analysis of effect of I/R and CP on apoptosis and caspase family members and PARP and (d) proteins related to the ER stress pathway. Relative protein expression was analyzed by band gray degree. β-actin is a normalization control (e). Six independent experiments were performed for above studies.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4645123&req=5

f2: CST pretreatment reduced ER-stress and decreased apoptosis.(a) Cleaved caspase-3 immunofluorescence staining and nuclear staining; apoptotic cells were stained in pink Scale bar = 100 μM. (b) Apoptosis cell numbers were counted in five different field of vision. (c) Western blot analysis of effect of I/R and CP on apoptosis and caspase family members and PARP and (d) proteins related to the ER stress pathway. Relative protein expression was analyzed by band gray degree. β-actin is a normalization control (e). Six independent experiments were performed for above studies.
Mentions: We used cleaved caspase-3 (a marker of cell apoptosis) antibody immunofluorescence staining to assess cell apoptosis. I/R increased the number of apoptotic cardiomyocytes, and CP significantly reduced the cell apoptosis (Fig. 2a,b). Consistently, I/R increased apoptosis markers (caspase-9, 7 and -3 cleavage, and poly (ADP-ribose) polymerase (PARP) cleaved into 89- and 24-kDa segments). CP lowered cleaved caspase-9, -7, -3 and PARP (Fig. 2c,e) for a role in reduced apoptosis.

Bottom Line: Cardiomyocytes endogenously produced CST and its expression was reduced after I/R.CST pretreatment decreased apoptosis especially endoplasmic reticulum (ER) stress response during I/R.The protective effects of CST were blocked by extracellular signal-regulated kinases 1/2 (ERK1/2) inhibitor PD90895 and phosphoinositide 3-kinase (PI3 K) inhibitor wortmannin.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology and Pathophysiology, School of Basic Medical Science, Peking University, P.R. China.

ABSTRACT
Catestatin (CST) is a catecholamine secretion inhibiting peptide as non-competitive inhibitor of nicotinic acetylcholine receptor. CST play a protective role in cardiac ischemia/reperfusion (I/R) but the molecular mechanism remains unclear. Cardiomyocytes endogenously produced CST and its expression was reduced after I/R. CST pretreatment decreased apoptosis especially endoplasmic reticulum (ER) stress response during I/R. The protection of CST was confirmed in H9c2 cardiomyoblasts under Anoxia/reoxygenation (A/R). In contrast, siRNA-mediated knockdown of CST exaggerated ER stress induced apoptosis. The protective effects of CST were blocked by extracellular signal-regulated kinases 1/2 (ERK1/2) inhibitor PD90895 and phosphoinositide 3-kinase (PI3 K) inhibitor wortmannin. CST also increased ERK1/2 and protein kinase B (Akt) phosphorylation and which was blocked by atropine and selective type 2 muscarinic acetylcholine (M2) receptor, but not type 1 muscarinic acetylcholine (M1) receptor antagonist. Receptor binding assay revealed that CST competitively bound to the M2 receptor with a 50% inhibitory concentration of 25.7 nM. Accordingly, CST inhibited cellular cAMP stimulated by isoproterenol or forskolin, and which was blocked by selective M2 receptor antagonist. Our findings revealed that CST binds to M2 receptor, then activates ERK1/2 and PI3 K/Akt pathway to inhibit ER stress-induced cell apoptosis resulting in attenuation cardiac I/R injury.

No MeSH data available.


Related in: MedlinePlus