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METTL3 Modulates TXNIP Expression to Affect the Activation of NLRP3 Inflammasome in Hepatic Cells Under Oxygen–Glucose Deprivation/Reperfusion Injury

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Abstract

Hepatic ischemia–reperfusion (I/R) injury is still a major risk factor and unsolved problem in hepatic surgery. Methyltransferase-like 3 (METTL3), an important m6A-modified methylase, regulates inflammation and cellular stress response. In this study, we demonstrated the special role of METTL3 and its underlying mechanism in hepatic I/R injury. In the mouse model of hepatic I/R and in the oxygen–glucose deprivation and reoxygenation (OGD/R)-induced AML12 and NCTC 1469 cells, the expression of METTL3 was significantly upregulated. Inhibition of METTL3 in OGD/R-induced AML12 and NCTC 1469 cells both increased the cell viability, declined the cell apoptosis, and decreased the reactive oxygen species (ROS) and the release levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18), diminishing NLRP3 and Caspase1-p20 expressions. Moreover, METTL3 positively modulated TXNIP expression in an m6A manner. TXNIP overexpression reversed the effects of METTL3 knockdown on OGD/R-induced injury in AML12 cells. Furthermore, inhibition of NLRP3 inflammasome activity contributed to the protective effects of TXNIP knockdown in OGD/R-induced AML12 cells. In conclusion, METTL3 knockdown alleviated OGD/R-induced hepatocyte injury, and the specific mechanism was associated with the inhibition of NLRP3 inflammasome activation, which was attributed to the reduction of TXNIP in an m6A-dependent manner.

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Funding

This work was supported by the Shaanxi Provincial Health Research Fund (No. 2022D005) and the National Natural Science Foundation of China (No. 82103553).

Shaanxi Provincial Health Research Fund,2022D005,National Natural Science Foundation of China,82103553

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Authors and Affiliations

  1. Anesthesia Department, the Second Affiliated Hospital of Xi’an Jiaotong University, No. 157, West 5th Road, Xi’an, Shaanxi Province, 710004, China

    Yong Zhang, Jianrui Lv, Gang Wu, Xiaoming Lei, Wei Li & Zhenni Zhang

  2. Department of General Surgery, Xuanwu Hospital, Capital Medical University, Beijing, China

    Jian Bai

  3. Department of General Practice, Beijing Friendship Hospital, Capital Medical University, Beijing, China

    Xue Zhang

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Contributions

Yong Zhang, Jianrui Lv, Jian Bai, Xue Zhang, Gang Wu, Xiaoming Lei, Wei Li, and Zhenni Zhang contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Yong Zhang, Jianrui Lv, Jian Bai, and Xue Zhang. Yong Zhang, Jianrui Lv, Gang Wu, Xiaoming Lei, and Wei Li analyzed experimental results. Yong Zhang, Jianrui Lv, Jian Bai, and Zhenni Zhang were responsible for data curation. The first draft of the manuscript was written by Yong Zhang and Jianrui Lv. Yong Zhang, Jianrui Lv, Jian Bai, Xue Zhang, Gang Wu, Xiaoming Lei, Wei Li, and Zhenni Zhang commented on previous versions of the manuscript. Yong Zhang, Jianrui Lv, Jian Bai, Xue Zhang, and Zhenni Zhang edited and revised the manuscript. All authors read and approved the final manuscript.

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10753_2023_1958_MOESM1_ESM.tif

Supplementary file1: METTL3 is upregulated in OGD/R-induced NCTC 1469 cells. (a) The mRNA level of METTL3 in NCTC 1469 cells treated with or without OGD/R. (b) The protein level of METTL3 in NCTC 1469 cells treated with or without OGD/R. *P < 0.05, **P < 0.01, ***P < 0.001. (TIF 542 KB)

10753_2023_1958_MOESM2_ESM.tif

Supplementary file2: METTL3 knockdown alleviates OGD/R-induced hepatic cell injury in NCTC 1469 cells. NCTC 1469 cells infected with sh-NC or sh-METTL3 lentivirus were subjected to OGD/R injury. The mRNA (a) and protein (b) expression levels of METTL3 were detected by qPCR and western blot, respectively. (c) Cell viability was assessed by CCK-8 assay. (d and e) The release levels of IL-1β and IL-18 were measured by ELISA assay. (f) The protein expression levels of NLRP3 and Caspase1-p20 were detected by western blot. ns indicates no statistical significance. *P < 0.05, **P < 0.01, ***P < 0.001. (TIF 1808 KB)

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Zhang, Y., Lv, J., Bai, J. et al. METTL3 Modulates TXNIP Expression to Affect the Activation of NLRP3 Inflammasome in Hepatic Cells Under Oxygen–Glucose Deprivation/Reperfusion Injury. Inflammation (2024). https://doi.org/10.1007/s10753-023-01958-4

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