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Apoptosis

, Volume 22, Issue 1, pp 108–117 | Cite as

Adiponectin exerts cardioprotection against ischemia/reperfusion injury partially via calreticulin mediated anti-apoptotic and anti-oxidative actions

  • Yang Sun
  • Dajun Zhao
  • Yang Yang
  • Chao Gao
  • Xing Zhang
  • Zhiqiang Ma
  • Shuai Jiang
  • Lin Zhao
  • Wenhao Chen
  • Kai Ren
  • Wei YiEmail author
  • Feng GaoEmail author
Article

Abstract

The underlying mechanisms of cardioprotection of adiponectin (APN) against ischemia/reperfusion (I/R) injury remain largely unknown. The present study aimed to investigate whether calreticulin (CRT) mediated APN’s cardioprotection against I/R injury. We inhibited mice cardiac CRT expression via intra-myocardial injection of CRT SiRNA, performed transient LAD ligation, measured the cardiac function, apoptosis and oxidative stress to identify CRT’s effects on cardioprotective actions of APN against I/R injury in vivo. LDH release and expression of CRT were measured in neonatal cardiomyocytes (NCM) subjected to simulated I/R (SI/R) and APN. CRT specific SiRNA was also utilized in vitro. CRT inhibition partially blunted cardioprotection of APN against I/R injury (evidenced by left ventricular ejection fraction and myocardial infarct size). It also blunted APN’s function against I/R induced apoptosis and oxidative stress (evidenced by TUNEL positive staining and reactive oxygen species production). In addition, SI/R increased LDH release, and administration of APN attenuated SI/R-induced cell death significantly. However, neither SI/R nor APN altered CRT expression in NCM. Inhibition of CRT expression blunted cardioprotective action of APN against SI/R induced apoptotic events (evidenced by TUNEL positive staining, LDH release and Caspase 3 activity). Furthermore, CRT inhibition significantly blunted APN’s anti-oxidative action (evidenced by gp91phox expression and superoxide generation). However, CRT inhibition did not attenuate AMPK phosphorylation by APN administration in NCM. Therefore, these novel findings strongly indicate that APN exerts cardioprotective effects against I/R injury partially via CRT mediated anti-apoptotic and anti-oxidative actions.

Keywords

Adiponectin Apoptosis Calreticulin Myocardial ischemia/reperfusion Oxidative stress 

Notes

Acknowledgments

This work was supported by National Natural Science Foundation of China (81422004, 81500263, 81470480, 81470477, 81100137, 81100081), China Postdoctoral Science Foundation (2015M572681), National High-tech R&D Program of China (2014AA020514, 2015AA020919), and Technological New Star Program of Shaanxi Province (2014KJXX-56).

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.

Supplementary material

10495_2016_1304_MOESM1_ESM.tif (55.7 mb)
Supplementary material 1 Supplementary Figure 1. Stealth RNAi (20μl; 0.8μg/g) or negative scramble SiRNA were injected into each mouse’s intramyocardia. (A-B) Efficiency of CRT inhibition in vivo determined by representative western blot and RT-PCR. **P <0.01 vs. Scramble SiRNA group. N=6-8. (B) The viability of normal NCM subjected to the SiRNA was assessed by performing an MTT assay, and the viability was expressed as an OD value. N=6-8 (TIF 57035 KB)
10495_2016_1304_MOESM2_ESM.tif (48.7 mb)
Supplementary material 2 Supplementary Figure 2. CRT expression was determined by representative western blots. *P <0.05, **P <0.01 vs. Control group. N=6-8 (TIF 49885 KB)

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Yang Sun
    • 1
    • 2
    • 3
  • Dajun Zhao
    • 2
  • Yang Yang
    • 4
  • Chao Gao
    • 5
  • Xing Zhang
    • 1
  • Zhiqiang Ma
    • 6
  • Shuai Jiang
    • 1
  • Lin Zhao
    • 2
  • Wenhao Chen
    • 2
  • Kai Ren
    • 2
  • Wei Yi
    • 2
    Email author
  • Feng Gao
    • 1
    Email author
  1. 1.Department of Aerospace MedicineFourth Military Medical UniversityXi’anChina
  2. 2.Department of Cardiovascular SurgeryXijing Hospital, Fourth Military Medical UniversityXi’anChina
  3. 3.Department of GeriatricXijing Hospital, Fourth Military Medical UniversityXi’anChina
  4. 4.Department of Biomedical EngineeringThe Fourth Military Medical UniversityXi’anChina
  5. 5.Department of CardiologyXijing Hospital, Fourth Military Medical UniversityXi’anChina
  6. 6.Department of Thoracic Surgery, Tangdu HospitalThe Fourth Military Medical UniversityXi’anChina

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