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MLKL Aggravates Ox-LDL-Induced Cell Pyroptosis via Activation of NLRP3 Inflammasome in Human Umbilical Vein Endothelial Cells

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Abstract

Atherosclerosis is a progressive chronic inflammation in the arterial walls. It is believed that the deposition of low-density lipoprotein (LDL) and its damage to endothelial cells play a vital role in atherosclerosis. Oxidized LDL (Ox-LDL) was confirmed to induce endothelial cell pyroptosis which plays an important role in intima inflammation and the development of atherosclerosis, but the underlying molecular mechanism needs to be explored. Here, we showed that ox-LDL upregulated the expression of mixed lineage kinase domain-like (MLKL) protein at both the mRNA and protein levels in endothelial cells, associated with the augment of pro-caspase-1 cleavage, interleukin-1β (IL-1β) maturation, pro-IL-1β production, and lactate dehydrogenase (LDH) release. Overexpression of MLKL substantially aggravated ox-LDL-induced increasing levels of caspase-1, IL-1β, pro-IL-1β, and LDH. MLKL-induced caspase-1 activation and IL-1β maturation were abolished by NLR family, pyrin domain-containing 3 (NLRP3) specific inhibitor MCC950, or extracellular high potassium concentration. Our findings indicated that MLKL is essential for regulation of ox-LDL-induced pyroptosis and inflammation through the activation of NLRP3 inflammasome, and suggested that MLKL could act as potential therapeutic targets to ameliorate atherosclerosis-related diseases.

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Funding

This work was supported by the National Natural Sciences Foundation of China (grant numbers 81871701 and 81772244), the Natural Science Fund of Guangdong (grant numbers 2020B1515020013, 2017A030313532 and 2017A030313535), and the Science and Technology Program of Guangzhou (grant number 201704020213).

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  1. Qian Wu and Xin He contributed equally to this work.

Authors and Affiliations

  1. Laboratory Medicine Center, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China

    Qian Wu, Xin He, Li-Mei Wu, Ru-Yi Zhang, Li-Min Li, Chang-Meng Wu, Yuan-Bin Lu, Bing Hu, Chao Shi, Zhi-Feng Lu, Biao Yang, Lei Zheng, Yan-Wei Hu & Qian Wang

  2. Laboratory Medicine Center, Zhujiang Hospital, Southern Medical University, Guangzhou, 510515, China

    Xin He, Li-Min Li & Qian Wang

  3. Department of Clinical Laboratory, Guangzhou Twelfth People’s Hospital, Guangzhou, 510620, China

    Li-Mei Wu

  4. Department of Clinical Laboratory, Guangzhou Women & Children Medical Center, Guangzhou Medical University, Guangzhou, China

    Yan-Wei Hu

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  1. Qian Wu
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Contributions

All authors contributed to the study conception and design. Qian Wu and Xin He designed the study, performed most part of the experiments, analyzed and interpreted the data, and wrote the manuscript. Li-Mei Wu, Ru-Yi Zhang, and Li-Min Li performed part of the experimental procedures. Chang-Meng Wu, Yuan-Bin Lu, Bing Hu, Chao Shi, Zhi-Feng Lu, and Biao Yang contributed to data acquisition. Lei Zheng, Yan-Wei Hu, and Qian Wang provided financial support and guided the completion of the project. All authors read and approved the final manuscript.

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Correspondence to Yan-Wei Hu or Qian Wang.

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Wu, Q., He, X., Wu, LM. et al. MLKL Aggravates Ox-LDL-Induced Cell Pyroptosis via Activation of NLRP3 Inflammasome in Human Umbilical Vein Endothelial Cells. Inflammation 43, 2222–2231 (2020). https://doi.org/10.1007/s10753-020-01289-8

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