Puerarin enhances TFEB-mediated autophagy and attenuates ROS-induced pyroptosis after ischemic injury of random-pattern skin flaps

Yingying Lai; Ningning Yang; Donghao Shi; Xianhui Ma; Yingying Huang; Jingzhou Lu; Xuzi Zhang; Hui Zhou; Weiyang Gao; Cong Mao; Long Wang

doi:10.1016/j.ejphar.2024.176621

Puerarin enhances TFEB-mediated autophagy and attenuates ROS-induced pyroptosis after ischemic injury of random-pattern skin flaps

Eur J Pharmacol. 2024 Apr 26:974:176621. doi: 10.1016/j.ejphar.2024.176621. Online ahead of print.

Authors

Yingying Lai¹, Ningning Yang¹, Donghao Shi², Xianhui Ma¹, Yingying Huang¹, Jingzhou Lu¹, Xuzi Zhang¹, Hui Zhou³, Weiyang Gao⁴, Cong Mao⁵, Long Wang⁶

Affiliations

¹ Department of Orthopaedics, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325027, China; Key Laboratory of Orthopaedics of Zhejiang Province, Wenzhou, 325027, China; The Second School of Medicine, Wenzhou Medical University, Wenzhou, 325027, China.
² The Second School of Medicine, Wenzhou Medical University, Wenzhou, 325027, China.
³ Graduate School, Jiangxi University of Traditional Chinese Medicine, Nanchang, 330002, China.
⁴ Department of Orthopaedics, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325027, China; Key Laboratory of Orthopaedics of Zhejiang Province, Wenzhou, 325027, China; The Second School of Medicine, Wenzhou Medical University, Wenzhou, 325027, China. Electronic address: weiyanggaoi@126.com.
⁵ Department of Orthopaedics, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325027, China; Key Laboratory of Orthopaedics of Zhejiang Province, Wenzhou, 325027, China; The Second School of Medicine, Wenzhou Medical University, Wenzhou, 325027, China. Electronic address: maocong@wmu.edu.cn.
⁶ Department of Orthopaedics, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325027, China; Key Laboratory of Orthopaedics of Zhejiang Province, Wenzhou, 325027, China; The Second School of Medicine, Wenzhou Medical University, Wenzhou, 325027, China. Electronic address: wanglongi@yeah.net.

PMID: 38679118
DOI: 10.1016/j.ejphar.2024.176621

Abstract

Background and aim: Necrosis of random-pattern flaps restricts their application in clinical practice. Puerarin has come into focus due to its promising therapeutic effects in ischemic diseases. Here, we employed Puerarin and investigated its role and potential mechanisms in flap survival.

Experimental procedure: The effect of Puerarin on the viability of human umbilical vein endothelial cells (HUVECs) was assessed by CCK-8, EdU staining, migration, and scratch assays. Survival area measurement and laser Doppler blood flow (LDBF) were utilized to assess the viability of ischemic injury flaps. Levels of molecules related to oxidative stress, pyroptosis, autophagy, transcription factor EB (TFEB), and the AMPK-TRPML1-Calcineurin signaling pathway were detected using western blotting, immunofluorescence, dihydroethidium (DHE) staining, RT-qPCR and Elisa.

Key results: The findings demonstrated that Puerarin enhanced the survivability of ischemic flaps. Autophagy, oxidative stress, and pyroptosis were implicated in the ability of Puerarin in improving flap survival. Increased autophagic flux and augmented tolerance to oxidative stress contribute to Puerarin's suppression of pyroptosis. Additionally, Puerarin modulated the activity of TFEB through the AMPK-TRPML1-Calcineurin signaling pathway, thereby enhancing autophagic flux.

Conclusions and implications: Puerarin promoted flap survival from ischemic injury through upregulation of TFEB-mediated autophagy and inhibition of oxidative stress. Our findings offered valuable support for the clinical application of Puerarin in the treatment of ischemic diseases, including random-pattern flaps.

Keywords: Autophagy; Oxidative stress; Puerarin; Pyroptosis; Random skin flaps.