Normothermic ex vivo Heart Perfusion Combined With Melatonin Enhances Myocardial Protection in Rat Donation After Circulatory Death Hearts via Inhibiting NLRP3 Inflammasome-Mediated Pyroptosis

Jun Lu; Liwei Xu; Zifeng Zeng; Chuqing Xue; Jiale Li; Xiong Chen; Pengyu Zhou; Shaoyan Lin; Yuhui Liao; Xianjin Du; Ronghua Yang; Shaoyi Zheng

doi:10.3389/fcell.2021.733183

Normothermic ex vivo Heart Perfusion Combined With Melatonin Enhances Myocardial Protection in Rat Donation After Circulatory Death Hearts via Inhibiting NLRP3 Inflammasome-Mediated Pyroptosis

Front Cell Dev Biol. 2021 Aug 31:9:733183. doi: 10.3389/fcell.2021.733183. eCollection 2021.

Authors

Jun Lu¹, Liwei Xu¹, Zifeng Zeng¹, Chuqing Xue¹, Jiale Li¹, Xiong Chen¹, Pengyu Zhou¹, Shaoyan Lin¹, Yuhui Liao², Xianjin Du³, Ronghua Yang⁴, Shaoyi Zheng¹

Affiliations

¹ Department of Cardiovascular Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, China.
² Molecular Diagnosis and Treatment Center for Infectious Diseases, Dermatology Hospital, Southern Medical University, Guangzhou, China.
³ Department of Critical Care Medicine, Renmin Hospital of Wuhan University, Wuhan, China.
⁴ Department of Burn Surgery, The First People's Hospital of Foshan, Foshan, China.

Abstract

Objective: The adoption of hearts from donation after circulatory death (DCD) is a promising approach for the shortage of suitable organs in heart transplantation. However, DCD hearts suffer from serious ischemia/reperfusion injury (IRI). Recent studies demonstrate that nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome-mediated pyroptosis is a novel target to ameliorate myocardial IRI. Melatonin is shown to inhibit NLRP3 inflammasome-mediated pyroptosis. Therefore, this study is designed to verify the hypothesis that melatonin can protect the heart graft preserved with ex vivo heart perfusion (EVHP) against myocardial IRI via inhibiting NLRP3 inflammasome-mediated pyroptosis in a rat model of DCD.

Methods: Donor-heart rats were randomly divided into three groups: (1) Control group: non-DCD hearts were harvested from heart-beating rats and immediately preserved with allogenic blood-based perfusate at constant flow for 105 min in the normothermic EVHP system; (2) DCD-vehicle group; and (3) DCD-melatonin group: rats were subjected to the DCD procedure with 25 min of warm ischemia injury and preserved by the normothermic EVHP system for 105 min. Melatonin (200 μmol/L) or vehicle was perfused in the cardioplegia and throughout the whole EVHP period. Cardiac functional assessment was performed every 30 min during EVHP. The level of oxidative stress, inflammatory response, apoptosis, and NLRP3 inflammasome-mediated pyroptosis of heart grafts submitted to EVHP were evaluated.

Results: Twenty five-minute warm ischemia injury resulted in a significant decrease in the developed pressure (DP), dP/dt _max , and dP/dt _min of left ventricular of the DCD hearts, while the treatment with melatonin significantly increased the DP, dP/dt _max of the left ventricular of DCD hearts compared with DCD-vehicle group. Furthermore, warm ischemia injury led to a significant increase in the level of oxidative stress, inflammatory response, apoptosis, and NLRP3 inflammasome-mediated pyroptosis in the hearts preserved with EVHP. However, melatonin added in the cardioplegia and throughout the EVHP period significantly attenuated the level of oxidative stress, inflammatory response, apoptosis, and NLRP3 inflammasome-mediated pyroptosis compared with DCD-vehicle group.

Conclusion: EVHP combined with melatonin post-conditioning attenuates myocardial IRI in DCD hearts by inhibiting NLRP3 inflammasome-mediated pyroptosis, which might expand the donor pool by the adoption of transplantable DCD hearts.

Keywords: donation after circulatory death; heart preservation; ischemia/reperfusion injury; melatonin; normothermic ex vivo heart perfusion; pyroptosis.