High uric acid caused pyroptosis of endothelial cells through the TXNIP/NLRP3 pathway

doi:10.3877/cma.j.issn.2095-3216.2021.02.005

Abstract

Abstract:

Objective

Hyperuricemia can lead to the activation of NLRP3 inflammasomes, resulting in the dysfunction and injury of endothelial cells, but the mechanism is unclear. This study aimed to explore whether uric acid (UA) could affect the epigenetic regulation and activation of inflammasomes through reactive oxygen species (ROS), leading to the endothelial damage.

Methods

Human umbilical vein endothelial cells (HUVEC) were cultured with UA (600 μmol/L) for 24 hours. Western blot was used to determine expression changes of pyroptosis-related proteins including NLRP3, caspase-1, and GSDMD-N. The levels of LDH, IL-1β, and IL-18 in the cells supernatant were detected by ELISA. The gain-of-function and loss-of-function experiments were performed to verify the role of NLRP3 inflammasomes in the pyroptosis of endothelial cells induced by high uric acid. Immunoprecipitation method was used to detect the interaction among oxidative stress-related proteins of TXNIP, TRX, and NLRP3 in the high uric acid environment. Overexpression and knock-down of TXNIP with lentivirus were used to further verify the regulation of NLRP3 by TXNIP, as well as its effect on pyroptosis of endothelial cells.

Results

The expression of pyroptosis-related proteins as NLRP3, caspase-1, and GSDMD-N in HUVEC treated with UA was increased, while the release of LDH, IL-1β, and IL-18 in the cells supernatant was also increased, with the proliferation of HUVEC being significantly inhibited. Gain-of-function and loss-of-function experiments confirmed that high uric acid led to pyroptosis of endothelial cells via NLRP3 inflammasomes. Treating the HUVEC with the ROS scavenger mito-Tempo inhibited the dissociation of TXNIP/TRX, thereby blocked the interaction between TXNIP and NLRP3, eventually resulting in inhibiting both the activation of NLRP3 inflammasomes and pyroptosis. Overexpression of TXNIP led to the activation of NLRP3 and subsequent pyroptosis, while knock-down of TXNIP caused the opposite results.

Conclusion

This study demonstrated that high uric acid caused inflammation and subsequent pyroptosis of the endothelial cells through ROS promoting the dissociation of TXNIP/TRX and then activating the NLRP3 inflammasomes.

Key words: ROS, TXNIP, NLRP3 inflammasome, Pyroptosis

Kun Chi, Zhangning Fu, Chengcheng Song, Xiaodong Geng, Chao Liu, Guangyan Cai, Xiangmei Chen, Quan Hong. High uric acid caused pyroptosis of endothelial cells through the TXNIP/NLRP3 pathway[J]. Chinese Journal of Kidney Disease Investigation(Electronic Edition), 2021, 10(02): 81-89.

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URL: https://zhsbyjdzzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-3216.2021.02.005

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Figures/Tables 6

References 27

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