Protective action of aspirin and its against endothelial Nlrp3 inflammasome activation in response to LPS stimuli
10.3867/j.issn.1000-3002.2017.10.059
- Author:
ZHOU XING
1
;
ZHONG YI-LIN
;
WU YAN-JIAO
;
HUANG YI
;
CHEN YANG
Author Information
1. School of Pharmaceutical
- Keywords:
Asporin;
Nlrp3 inflammasome;
CAECs;
ROS
- From:
Chinese Journal of Pharmacology and Toxicology
2017;31(10):984-984
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE Recent studies have demonstrated that the Nlrp3 inflammasome serve as a central role in the pathogenesis of cardiovascular diseases and endothelial dysfunction occurs in association with several cardiovascular risk factors. Given the demonstrated anti-inflammatory effects of aspirin, the present study was designed to test whether aspirin diminish NLRP3 inflammasome activation and prevent endothelium injury and associated coronary artery damage during LPS. METHODS Mouse carotid arterial endothelial cells (CAECs) were cultured and treated with 0.1-3 mmol·L-1 of aspirin in response to LPS (2 μg·mL-1) stimuli. After 24 h, the Nlrp3 inflammasome complexes consist of varied proteins were analyzed by WB. NO and T-AOC in the supernatant was detected by ELISA. Intracellular reactive oxygen species (ROS) generation for 24 h was observed by DCF fluorescence. The mice were treated with aspirin (12.5 mg·kg-1 per day, 62.5 mg·kg-1 per day, 125 mg·kg-1 per day) and dexametha?sone (0.0182 mg · kg- 1 per day) for 7 d. The level of IL- 1β,IL- 18 protein was detected by ELISA. RESULTS Immunofluorescence results showed the colocalization of Nlrp3 with ASC or caspase 1 decrease in a concentration- dependent manner. Meanwhile, the expression of Nlrp3 and caspase 1 protein was decreased with the concentration of aspirin, but no changes the expression of ASC protein. Nlrp3 protein levels in CAECs were 0.33- 0.8- fold and cle- caspase 1 protein levels in CAECs were 0.48-1-fold compared to those in LPS stimulation when treated with 0.1-3 mmol·L-1 aspirin for 24 h (P<0.01). Aspirin significantly antagonized the effect of LPS on NO (1.22-1.91-fold that of LPS stimulation, P<0.01) and T-AOC expression (1.02-1.90-fold that of LPS stimulation, P<0.01). As the different concentration of aspirin treated, the generation of ROS was 0.51-1.10-fold that of LPS stimulation (P<0.01). In vivo data shown the level of IL-1β, IL-18 protein from serum are in concordance with the level of Nlrp3 inflammasome activation. CONCLUSION We conclude that aspirin has anti- inflammatory properties, protecting CAECs from LPS-induced injury by inhibition of NLRP3 inflammasome activation through ROS pathway.
