2. Academic Validation
  3. DTX3L induced NLRP3 ubiquitination inhibit R28 cell pyroptosis in OGD/R injury

DTX3L induced NLRP3 ubiquitination inhibit R28 cell pyroptosis in OGD/R injury

  • Biochim Biophys Acta Mol Cell Res. 2023 Jan 24;1870(3):119433. doi: 10.1016/j.bbamcr.2023.119433.
Ziyu Zhou 1,  Lei Shang 2,  Qi Zhang 3,  Ximin Hu 4,  Ju-Fang Huang 5,  Kun Xiong 6
Affiliations

Affiliations

  • 1 Department of Anatomy and Neurobiology, School of Basic Medical Science, Central South University, Changsha 410008, China; The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Institute of Basic Medicine and Cancer (IBMC), Chinese Academy of Sciences, Hangzhou 310022, China.
  • 2 Jiangxi Research Institute of Ophthalmology and Visual Sciences, Affiliated Eye Hospital of Nanchang University, Nanchang 330006, China. Electronic address: shanglei1986@ncu.edu.cn.
  • 3 Department of Anatomy and Neurobiology, School of Basic Medical Science, Central South University, Changsha 410008, China. Electronic address: zhangqi2014@csu.edu.cn.
  • 4 Department of Anatomy and Neurobiology, School of Basic Medical Science, Central South University, Changsha 410008, China.
  • 5 Department of Anatomy and Neurobiology, School of Basic Medical Science, Central South University, Changsha 410008, China. Electronic address: huangjufang@csu.edu.cn.
  • 6 Department of Anatomy and Neurobiology, School of Basic Medical Science, Central South University, Changsha 410008, China; Hunan Key Laboratory of Ophthalmology, Changsha 410008, China; Key Laboratory of Emergency and Trauma, Ministry of Education, College of Emergency and Trauma, Hainan Medical University, Haikou 571199, China. Electronic address: xiongkun2001@163.com.
PMID: 36706922 DOI: 10.1016/j.bbamcr.2023.119433
Abstract

Ischemia/reperfusion (I/R) injury is one of the most common etiologies in many diseases. Retinal I/R leads to cytokine storm, resulting in tissue damage and cell death. Pyroptosis, a novel type of regulated cell death, occurs after cellular I/R injury. In this study, we established an oxygen glucose deprivation (OGD/R) cellular model (R28) to simulate retinal I/R injury. We conducted an LDH assay, and EthD-III and PI staining procedures to confirm Pyroptosis. Mass spectrometry and bioinformatics analysis were used to identify the possible proteins interacting with NLRP3. Co-IP and various Molecular Biology techniques were used to investigate the possible modes regulating NLRP3 by DTX3L. EthD-III, PI staining and LDH assays demonstrated Pyroptosis induced by OGD/R injury, mediated via NLRP3 pathway. Mass spectrometry and bioinformatics analysis screened out three candidate proteins interacting with NLRP3, and further Co-IP experiment indicated that DTX-3L may interact with NLRP3 to regulate its protein levels after injury. Co-IP experiments and various Molecular Biology methods demonstrated that DTX3L ubiquitinates NLRP3 resulting in Pyroptosis after R28 OGD/R injury. Further, NLRP3 LRR and DTX3L RING domains interact with each Other. Our study demonstrated that DTX3L may ubiquitinate NLRP3 to regulate OGD/R-induced Pyroptosis globally in R28 cells.

Keywords

DTX3L; NLRP3; OGD/R; Pyroptosis; Ubiquitination.

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