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Trimetazidine affects pyroptosis by targeting GSDMD in myocardial ischemia/reperfusion injury

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Abstract

Objective

Trimetazidine (TMZ) exerts a strong inhibitory effect on ischemia/reperfusion (I/R) injury. Inflammation plays a key role in I/R injury. We hypothesized that TMZ may protect cardiomyocytes from I/R injury by inhibiting inflammation.

Methods

The left anterior descending coronary artery was ligated for 30 min followed by 6 h of reperfusion to establish a model of I/R injury. H9c2 cardiomyocytes were subjected to 2 h of hypoxia and 3 h of normoxic conditions to establish a model of hypoxia/reoxygenation (H/R) injury. We monitored the change in pyroptosis by performing Western blot analysis, microscopy and ELISA.

Results

I/R and H/R treatment stimulated gasdermin D-N domain (GSDMD-N) expression in cardiomyocytes (sham onefold vs. I/R 2.5-fold; control onefold vs. H/R 2.0-fold). Moreover, TMZ increased the viability of H9c2 cardiomyocytes subjected to H/R treatment (H/R 65.0% vs. H/R + TMZ 85.3%) and reduced the infarct size in vivo (I/R 47.0% vs. I/R + TMZ 28.3%). H/R and I/R treatment increased the levels of TLR4, MyD88, phospho-NF-κB p65 and the NLRP3 inflammasome; however, TMZ reduced the expression of these proteins. Additionally, TMZ inhibited noncanonical inflammasome signaling induced by I/R injury.

Conclusions

In summary, TMZ alleviated pyroptosis induced by myocardial I/R injury through the TLR4/MyD88/NF-κB/NLRP3 inflammasome pathway. Therefore, TMZ represents an alternative treatment for myocardial I/R injury.

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Abbreviations

AAR:

Area at risk

CCK-8:

Cell counting kit-8

CK-MB:

Creatine kinase-MB

GSDMD-FL:

Gasdermin D-full length

GSDMD-N:

Gasdermin D-N domain

HE:

Hematoxylin and eosin

H/R:

Hypoxia/reoxygenation

INF:

Infarct size

I/R:

Ischemia/reperfusion

LAD:

Left anterior descending

LDH:

Lactate dehydrogenase

LV:

Left ventricle

TMZ:

Trimetazidine

TTC:

2,3,5-Triphenyltetrazolium chloride

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Acknowledgements

The work was supported by the National Natural Science Foundation of China (Grant No. 81670227 and 81600341), the Natural Science Foundation of Zhejiang Province (Grant No. LQ19H020004), the Cardiac Rehabilitation and Metabolic Therapy research fund (Grant No. 2019ZJXQN03), the Medical Health Science and Technology Project of Zhejiang Provincial (Grant No. 2021KY1072), the Wenzhou Science and Technology Bureau (Grant No. Y20170045, Y20160030 and Y20180105) and the Traditional Chinese Medicine Administration of Zhejiang Province (Grant No. 2016ZA137).

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Author notes

  1. Xudong Chen and Shuang Lin have contributed equally to this work.

Authors and Affiliations

  1. Department of Cardiology, The Key Lab of Cardiovascular Disease of Wenzhou, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, People’s Republic of China

    Xudong Chen, Shuang Lin, Peiren Shan, Weiqi Wang, Zhouqing Huang, Bozhi Ye & Weijian Huang

  2. Department of Cardiology, Ningbo Hangzhou Bay Hospital, Ningbo, Zhejiang, People’s Republic of China

    Xudong Chen

  3. Department of Emergency, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, People’s Republic of China

    Shanshan Dai

  4. Department of Cardiology, The Second Affiliated Hospital of Jiaxing University, Jiaxing, Zhejiang, People’s Republic of China

    Jibo Han

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Correspondence to Bozhi Ye.

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11_2021_1530_MOESM1_ESM.tif

Supplementary file1 (TIF 1362 KB) Supplementary Fig. S1 Schematic of the protocol and structure of TMZ. (A) Schematic of the protocol for the I/R and H/R study. After different treatments, tissue or cell samples were collected for further experimentation. (B) Schematic representation of the groups and indicators. (C) The structure of TMZ used in this study. I/R, ischemia/reperfusion; TMZ, trimetazidine; control, normoxic; H/R, hypoxia/reoxygenation; CK-MB, creatine kinase-MB; TTC, 2,3,5-triphenyltetrazolium chloride; HE, hematoxylin-eosin; IHC, immunohistochemistry; WB, Western blot analysis; ELISA, enzyme-linked immunosorbent assay; LAD, left anterior descending; CCK-8, Cell Counting Kit-8; LDH, lactate dehydrogenase; IF, immunofluorescence

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Chen, X., Lin, S., Dai, S. et al. Trimetazidine affects pyroptosis by targeting GSDMD in myocardial ischemia/reperfusion injury. Inflamm. Res. 71, 227–241 (2022). https://doi.org/10.1007/s00011-021-01530-6

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