Effects of MgSO4 on inhibiting Nod-like receptor protein 3 inflammasome involve decreasing intracellular calcium

Ya-Ying Chang; Ming-Chang Kao; Jui-An Lin; Tsung-Ying Chen; Ching-Feng Cheng; Chih-Shung Wong; I-Shiang Tzeng; Chun-Jen Huang

doi:10.1016/j.jss.2017.09.005

Effects of MgSO₄ on inhibiting Nod-like receptor protein 3 inflammasome involve decreasing intracellular calcium

J Surg Res. 2018 Jan:221:257-265. doi: 10.1016/j.jss.2017.09.005. Epub 2017 Sep 30.

Authors

Ya-Ying Chang¹, Ming-Chang Kao², Jui-An Lin³, Tsung-Ying Chen⁴, Ching-Feng Cheng⁵, Chih-Shung Wong⁶, I-Shiang Tzeng⁷, Chun-Jen Huang⁸

Affiliations

¹ Institute of Medical Sciences, Tzu Chi University, Hualien, Taiwan; Department of Anesthesiology, Taipei Tzu Chi Hospital, Taipei, Taiwan; School of Medicine, Tzu Chi University, Hualien, Taiwan.
² Department of Anesthesiology, Taipei Tzu Chi Hospital, Taipei, Taiwan; School of Medicine, Tzu Chi University, Hualien, Taiwan.
³ Department of Anesthesiology, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan; School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
⁴ School of Medicine, Tzu Chi University, Hualien, Taiwan; Department of Anesthesiology, Buddhist Tzu Chi General Hospital, Hualien, Taiwan.
⁵ Institute of Medical Sciences, Tzu Chi University, Hualien, Taiwan; School of Medicine, Tzu Chi University, Hualien, Taiwan; Department of Pediatrics, Buddhist Tzu Chi General Hospital, Hualien, Taiwan.
⁶ Department of Anesthesiology, Cathay General Hospital, Taipei, Taiwan.
⁷ Department of Medical Research, Taipei Tzu Chi Hospital, Taipei, Taiwan.
⁸ Department of Anesthesiology, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan; Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan. Electronic address: huangcj1112@gmail.com.

PMID: 29229137
DOI: 10.1016/j.jss.2017.09.005

Abstract

Background: Nod-like receptor protein 3 (NLRP3) inflammasome is a multiprotein complex composed of NLRP3, caspase-1, and apoptosis-associated speck-like protein containing a caspase recruitment domain. Activation of NLRP3 inflammasome leads to interleukin-1β (IL-1β) upregulation and pyroptosis, a proinflammatory cell death characterized by increased cell size. Of note, calcium signaling is crucial for NLRP3 inflammasome activation. This study elucidated the effects of magnesium sulfate (MgSO₄), a potent calcium antagonist, on modulating NLRP3 inflammasome.

Materials and methods: THP-1 cells, the human monocytic leukemia cell line, were treated with lipopolysaccharide (LPS, 1 μg/ml) plus nigericin (5 μM) (the LPS + Nig group) and LPS plus nigericin plus MgSO₄ (20 mM) [the LPS + Nig + M(20)] to facilitate investigations. Levels of IL-1β, pyroptosis, and NLRP3 inflammasome induction as well as intracellular calcium were assayed.

Results: IL-1β concentration of the LPS + Nig + M(20) group was significantly lower than the LPS + Nig group (P = 0.001). Cell size of the LPS + Nig + M(20) group was significantly smaller than the LPS + Nig group (P < 0.001). Level of pyroptotic cell death of the LPS + Nig + M(20) group was significantly lower than the LPS + Nig group (P = 0.004). NLRP3 mRNA and protein concentrations of the LPS + Nig + M(20) group were also significantly lower than the LPS + Nig group (P = 0.021 and P < 0.001). Similarly, apoptosis-associated speck-like protein containing a caspase recruitment domain speck formation ratio and caspase-1 concentration of the LPS + Nig + M(20) group were significantly lower than the LPS + Nig group (both P < 0.001). The change in intracellular calcium level of the LPS + Nig + M(20) group was significantly smaller than the LPS + Nig group (P = 0.001).

Conclusions: MgSO₄ inhibits NLRP3 inflammasome, IL-1β upregulation, and pyroptosis. The mechanism is consistent with decreased intracellular calcium levels.

Keywords: ASC; Caspase-1; Magnesium sulfate; Pyroptosis; THP-1.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Calcium Signaling / drug effects*
Caspase 1 / metabolism
Caspase Activation and Recruitment Domain
Humans
Inflammasomes / antagonists & inhibitors*
Interleukin-1beta / metabolism
Magnesium Sulfate / pharmacology*
NLR Family, Pyrin Domain-Containing 3 Protein / metabolism*
Pyroptosis / drug effects*
THP-1 Cells

Substances

IL1B protein, human
Inflammasomes
Interleukin-1beta
NLR Family, Pyrin Domain-Containing 3 Protein
NLRP3 protein, human
Magnesium Sulfate
Caspase 1