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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

Catestatin (CST) pretreatment (CP) reduced infarct size and maintained cardiac function after cardiac ischemia-reperfusion (I/R) injury in rats.(a) The procedure of global cardiac I/R (n = 18) and CP (n = 18) and original recording of left ventricular (LV) pressure. (b) The ChgA protein and its cleavage fragments expression were measured by western blot. (c) Heart tissue was stained with triphenyltetrazolium chloride and infarct size (white color) was measured by volume (n = 6 for each group). Evaluation of cardiac function including LV end diastolic pressure (LVEDP)(b), LV developed pressure (dLVP, LV systolic pressure–diastolic pressure) (d), and LV ±dp/dtmax (e) during I/R and CP (total 36 rats were recruited). Data are mean ± SD. *P < 0.05, **P < 0.01 vs I/R group. #P < 0.05 vs parameters before ischemia.
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f1: Catestatin (CST) pretreatment (CP) reduced infarct size and maintained cardiac function after cardiac ischemia-reperfusion (I/R) injury in rats.(a) The procedure of global cardiac I/R (n = 18) and CP (n = 18) and original recording of left ventricular (LV) pressure. (b) The ChgA protein and its cleavage fragments expression were measured by western blot. (c) Heart tissue was stained with triphenyltetrazolium chloride and infarct size (white color) was measured by volume (n = 6 for each group). Evaluation of cardiac function including LV end diastolic pressure (LVEDP)(b), LV developed pressure (dLVP, LV systolic pressure–diastolic pressure) (d), and LV ±dp/dtmax (e) during I/R and CP (total 36 rats were recruited). Data are mean ± SD. *P < 0.05, **P < 0.01 vs I/R group. #P < 0.05 vs parameters before ischemia.

Mentions: Ischemia reperfusion procedure and CST pretreatment (CP) (CST supplementation for 15 min before I/R) were performed (Fig. 1a). After 30 min of ischemia and 1 h of reperfusion, cardiac endogenous ChgA mRNA (see supplementary Fig. S1 online) and protein expression were no changes, but CST fragments (21 kD and 7.5 kD) reduced comparison to control (Fig. 1b). Different dose of CST perfusion for 1 h did not induce lactate dehydrogenase (LDH) leakage (supplementary Fig. S2 online). CST (25 nM, 50 nM and 100 nM) pretreatment decreased LDH leakage and cardiac troponin I (cTNI) level in perfusate at different reperfusion time point (supplementary Fig. S3 online), which indicated that CST pretreatment lowered the ischemia reperfusion (I/R) injury, especially CST at 100 nM exhibited well protection. So we selected 100 nM CST in following experiments. As Fig. 1c showed, I/R caused global cardiac infarction, and CP significantly reduced infarct size as compared with I/R alone (9.8 ± 4.2% vs 27.5 ± 8.2%, P = 0.0001).


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)

Catestatin (CST) pretreatment (CP) reduced infarct size and maintained cardiac function after cardiac ischemia-reperfusion (I/R) injury in rats.(a) The procedure of global cardiac I/R (n = 18) and CP (n = 18) and original recording of left ventricular (LV) pressure. (b) The ChgA protein and its cleavage fragments expression were measured by western blot. (c) Heart tissue was stained with triphenyltetrazolium chloride and infarct size (white color) was measured by volume (n = 6 for each group). Evaluation of cardiac function including LV end diastolic pressure (LVEDP)(b), LV developed pressure (dLVP, LV systolic pressure–diastolic pressure) (d), and LV ±dp/dtmax (e) during I/R and CP (total 36 rats were recruited). Data are mean ± SD. *P < 0.05, **P < 0.01 vs I/R group. #P < 0.05 vs parameters before ischemia.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: Catestatin (CST) pretreatment (CP) reduced infarct size and maintained cardiac function after cardiac ischemia-reperfusion (I/R) injury in rats.(a) The procedure of global cardiac I/R (n = 18) and CP (n = 18) and original recording of left ventricular (LV) pressure. (b) The ChgA protein and its cleavage fragments expression were measured by western blot. (c) Heart tissue was stained with triphenyltetrazolium chloride and infarct size (white color) was measured by volume (n = 6 for each group). Evaluation of cardiac function including LV end diastolic pressure (LVEDP)(b), LV developed pressure (dLVP, LV systolic pressure–diastolic pressure) (d), and LV ±dp/dtmax (e) during I/R and CP (total 36 rats were recruited). Data are mean ± SD. *P < 0.05, **P < 0.01 vs I/R group. #P < 0.05 vs parameters before ischemia.
Mentions: Ischemia reperfusion procedure and CST pretreatment (CP) (CST supplementation for 15 min before I/R) were performed (Fig. 1a). After 30 min of ischemia and 1 h of reperfusion, cardiac endogenous ChgA mRNA (see supplementary Fig. S1 online) and protein expression were no changes, but CST fragments (21 kD and 7.5 kD) reduced comparison to control (Fig. 1b). Different dose of CST perfusion for 1 h did not induce lactate dehydrogenase (LDH) leakage (supplementary Fig. S2 online). CST (25 nM, 50 nM and 100 nM) pretreatment decreased LDH leakage and cardiac troponin I (cTNI) level in perfusate at different reperfusion time point (supplementary Fig. S3 online), which indicated that CST pretreatment lowered the ischemia reperfusion (I/R) injury, especially CST at 100 nM exhibited well protection. So we selected 100 nM CST in following experiments. As Fig. 1c showed, I/R caused global cardiac infarction, and CP significantly reduced infarct size as compared with I/R alone (9.8 ± 4.2% vs 27.5 ± 8.2%, P = 0.0001).

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