1. Academic Validation
  2. A novel mechanism of Dimethyl ester of Alpha-ketoglutarate in suppressing Paraquat-induced BEAS-2B cell injury by alleviating GSDME dependent pyroptosis

A novel mechanism of Dimethyl ester of Alpha-ketoglutarate in suppressing Paraquat-induced BEAS-2B cell injury by alleviating GSDME dependent pyroptosis

  • Phytomedicine. 2023 Feb 5;112:154698. doi: 10.1016/j.phymed.2023.154698.
Yahui Tang 1 Kaiyuan Chen 2 Zhong Xiao 2 Guangliang Hong 2 Lufeng Hu 3 Jianping Cai 4 Zhongqiu Lu 5
Affiliations

Affiliations

  • 1 Department of Emergency Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China; Wenzhou Key Laboratory of emergency and disaster medicine, Wenzhou 325000, China. Electronic address: tangyahui@wzhospital.cn.
  • 2 Department of Emergency Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China; Wenzhou Key Laboratory of emergency and disaster medicine, Wenzhou 325000, China.
  • 3 Wenzhou Key Laboratory of emergency and disaster medicine, Wenzhou 325000, China; Department of Pharmacy, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
  • 4 Ministry of Health Key Laboratory of Geriatrics (J.-P.C.), Beijing Hospital, National Center of Gerontology; Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, China.
  • 5 Department of Emergency Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China; Wenzhou Key Laboratory of emergency and disaster medicine, Wenzhou 325000, China. Electronic address: wzykdxlzq@126.com.
Abstract

Background: Acute lung injury (ALI) induced by paraquat (PQ) progresses rapidly, leading to high mortality; however, there is no specific antidote. Our limited knowledge of the pathogenic toxicological mechanisms of PQ has hindered the development of treatments against PQ exposure.

Purpose: Pyroptosis is a form of programmed cell death recently identified as a novel molecular mechanism adopted by chemotherapeutic drugs for Cancer therapy. However, the involvement of Pyroptosis in PQ-induced lung injury has not been reported. Therefore, we investigated the effects of PQ on the lung tissues to elucidate the molecular mechanisms underlying its toxicity, especially its ability to induce Pyroptosis.

Methods: To observe the morphological changes of BEAS-2B cells exposed to PQ, the plasma membrane damage of the cells was detected by LDH release assay, mitochondrial function and cell metabolism were detected by energy metabolism analysis. Western blotting was used to detect the protein levels of GSDMD, C-GSDMD, GSDME and N-GSDME in BEAS-2B cells. Metabolites of TCA cycle were detected by metabolomics, and the changes of TCA cycle metabolic enzymes in cells were detected by Western blotting.

Results: We observed that PQ induced proteolytic cleavage of gasdermin E (GSDME) with concomitant cleavage of Caspase 3 in BEAS-2B cells. Knockout of GSDME attenuated PQ-induced cell death. Additionally, PQ induced ROS accumulation, mitochondrial depolarisation, and mitochondrial dysfunction in these cells. PQ activated the Caspase 3/GSDME pathway and damaged the cytoplasmic membrane in cells, leading to Pyroptosis. We demonstrated that DMK suppressed PQ-induced Pyroptosis by blocking PQ-induced Caspase 3/GSDME pathway activation, reducing cellular ROS levels, and improving mitochondrial function.

Conclusion: These findings provide novel insights into the previously unrecognized mechanism of GSDME-dependent Pyroptosis in PQ poisoning.

Keywords

DMK; GSDME; Lung injury; Paraquat; Pyroptosis.

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