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The effects of curcumin post-treatment against myocardial ischemia and reperfusion by activation of the JAK2/STAT3 signaling pathway

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Abstract

In this study, we evaluated the effect of curcumin (Cur) post-treatment on isolated perfused rat hearts that had been subjected to a protocol of ischemia and reperfusion injury. We also examined whether the Janus kinase 2 and signal transducer and activator 3 of transcription (JAK2/STAT3) signaling pathway plays a role in the cardioprotective effects of Cur post-treatment. Isolated perfused rat hearts were subjected to 60 min of ischemia, followed by 60 min of reperfusion. The hearts were exposed to 1-μM Cur during the first 10 min of reperfusion in the absence or presence of the JAK kinase-specific inhibitor AG490 (AG, 1 μM). The Cur treatment conferred a cardioprotective effect, and the treated hearts demonstrated an improved post-ischemic cardiac functional recovery, a decreased myocardial infarct size and decreased lactate dehydrogenase release in the coronary flow, a reduced number of apoptotic cardiomyocytes, up-regulation of the anti-apoptotic protein Bcl2 and down-regulation of the pro-apoptotic protein Caspase3. AG blocked the Cur-mediated cardioprotection by inhibiting the JAK2/STAT3 signaling pathway, as reflected by the abrogation of the Cur-induced up-regulation of Bcl2 and down-regulation of Caspase3. The results suggest that Cur post-treatment can attenuate IR injury through the activation of the JAK2/STAT3 signaling pathway, which transmits a survival signal to the myocardium.

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Abbreviations

Cur:

Curcumin

AG, AG490:

A specific inhibitor of the JAK2/STAT3 signaling pathway

IR:

Ischemia and reperfusion

JAK2/STAT3:

Janus kinase 2 and signal transducer and activator 3 of transcription protein

LVEDP:

Left ventricular end-diastolic pressure

LVDP:

Left ventricular peak develo** pressure

HR:

Heart rate

CF:

Coronary flow, the volume of buffer that overflowed out of an isolated heart chamber during a 1 min interval

DP:

The index of cardiac contractile function = (HR × LVDP)/1,000

+dP/dt max:

The maximum rate of pressure change in the ventricle

TUNEL:

Terminal deoxynucleotidyl transferase dUTP nick end labeling assay

DAPI:

4′, 6-diamidino-2-phenylindole

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Acknowledgments

This study was supported by grants from the National Natural Science Foundation of China (81102687) and the Academic Promotion Project of ****g Hospital (XJZT09M16 and XJZT10M12).

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The authors have no disclosures to declare.

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Authors and Affiliations

  1. Department of Cardiovascular Surgery, ****g Hospital, The Fourth Military Medical University, 127 Changle West Road, **’an, 710032, China

    Weixun Duan, Yang Yang, Shiqiang Yu, **cheng Liu, Zhenxiao ** & Dinghua Yi

  2. Department of Prosthodontics, School of Stomatology, The Fourth Military Medical University, 145 Changle West Road, **’an, 710032, China

    Juanjuan Yan

  3. Department of Physiology and Cardiology, ****g Hospital, The Fourth Military Medical University, **’an, 710032, China

    **gjun Zhou & Jianying Zhang

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  1. Weixun Duan
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  2. Yang Yang
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  5. **cheng Liu
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  8. Zhenxiao **
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Correspondence to Zhenxiao ** or Dinghua Yi.

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W. Duan and Y. Yang contributed equally to this work.

Electronic supplementary material

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395_2012_263_MOESM1_ESM.docx

Supplement Tab. 1 The effects of Cur and AG on the hemodynamic parameters of isolated rat hearts. The results are expressed as the mean ± SEM of 8 rats per group. **P<0.01 vs. the control group, ##P<0.01 vs. the Cur group. Cur, curcumin; AG, AG490; HR, heart rate; LVDP, left ventricular peak develo** pressure; +dP/dt max, the maximum rate of pressure change in the ventricle; DP, the index of cardiac contractile function as determined using the product of HR × LVDP/1000; CF, coronary flow (DOCX 16 kb)

395_2012_263_MOESM2_ESM.docx

Supplement Fig. 1 Experimental protocol. All of the hearts were initially perfused and stabilized with KHB for 10 min. The control hearts continued to be perfused with KHB, whereas the hearts in the Cur group were perfused with KHB for 60 min, subjected to KHB with 1 μM Cur for 10 min, then perfused with KHB for 50 min. The hearts in the AG group were perfused with KHB for 60 min, subjected to KHB with 1 μM AG for 10 min, then perfused with KHB for 50 min. Cur, curcumin; AG, AG490; KHB, Krebs-Henseleit buffer (DOCX 38 kb)

395_2012_263_MOESM3_ESM.docx

Supplement Fig. 2 The effects of Cur and AG on the cardiac function of normal isolated rat hearts. Representative images of LVDP curves, as monitored by an MP150 pressure transducer system (Biopac, USA), are shown (n=8). Cur, curcumin; AG, AG490; LVDP, left ventricular peak develo** pressure (DOCX 119 kb)

395_2012_263_MOESM4_ESM.docx

Supplement Fig. 3 The effects of Cur and AG on the myocardial infarct size of normal isolated rat hearts. Representative images of the myocardial infarct size are shown. The infarction size is expressed as the percentage of infarct relative to the mass at risk. The results are expressed as the mean ± SEM of 8 rat hearts per group. There was no difference in the myocardial infarct size among the three groups. Cur, curcumin; AG, AG490 (DOCX 593 kb)

395_2012_263_MOESM5_ESM.docx

Supplement Fig. 4 The effects of Cur and AG on LDH release in normal isolated rat hearts. The LDH levels were determined using an ELISA kit according to the manufacturer’s instructions. The LDH release of the three groups is shown in the figure. The amount of LDH was normalized against the wet weight of the heart and is expressed as IU/g. The results are expressed as the mean ± SEM of 8 rat hearts per group. There was no difference in the LDH release among the three groups. Cur, curcumin; AG, AG490 (DOCX 33 kb)

395_2012_263_MOESM6_ESM.docx

Supplement Fig. 5 The effects of Cur and AG on apoptosis in normal isolated rat hearts. Representative images of apoptotic cardiomyocytes are shown. The apoptotic cells were detected by immunofluorescent staining with TUNEL (green), and DAPI (blue) staining was used to label the nuclei. The results are expressed as the mean ± SEM of 8 rat hearts per group. There was no difference in the amount of apoptosis among the three groups. Cur, curcumin; AG, AG490 (DOCX 246 kb)

395_2012_263_MOESM7_ESM.docx

Supplement Fig. 6 The effects of Cur and AG on JAK2 and STAT3 phosphorylation in normal isolated rat hearts. Representative images of the Western blot results are shown. The results are expressed as the mean ± SEM of 8 rat hearts per group. **P<0.01 vs. the control group, ##P<0.01 vs. the Cur group. Cur, curcumin; AG, AG490 (DOCX 146 kb)

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Duan, W., Yang, Y., Yan, J. et al. The effects of curcumin post-treatment against myocardial ischemia and reperfusion by activation of the JAK2/STAT3 signaling pathway. Basic Res Cardiol 107, 263 (2012). https://doi.org/10.1007/s00395-012-0263-7

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