Protective action of aspirin and its against endothelial Nlrp3 inflammasome activation in response to LPS stimuli

Xing ZHOU,Yi-lin ZHONG,Yan-jiao WU,Yi HUANG,Yang CHEN

(1.School of Pharmaceutical,Guangzhou University of Chinese Medicine,Guangzhou 510000,China;2.Department of Stomatology,The First Affiliated Hospital,Jinan University,Guangzhou 510630,China)

T1-46

Protective action of aspirin and its against endothelial Nlrp3 inflammasome activation in response to LPS stimuli

Xing ZHOU1,Yi-lin ZHONG1,Yan-jiao WU1,Yi HUANG2,Yang CHEN1

(1.School of Pharmaceutical,Guangzhou University of Chinese Medicine,Guangzhou 510000,China;2.Department of Stomatology,The First Affiliated Hospital,Jinan University,Guangzhou 510630,China)

OBJECTIVERecent 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.METHODSMouse carotid arterial endothelial cells(CAECs)were cultured and treated with 0.1-3 mmol·L-1of 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-1per day,62.5 mg·kg-1per day,125 mg·kg-1per day)and dexametha⁃sone(0.0182 mg·kg-1per day)for 7 d.The level of IL-1β，IL-18 protein was detected by ELISA.RESULTSImmunofluorescence 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-1aspirin 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.CONCLUSIONWe conclude that aspirin has anti-inflammatory properties,protecting CAECs from LPS-induced injury by inhibition of NLRP3 inflammasome activation through ROS pathway.

Asporin;Nlrp3 inflammasome;CAECs;ROS

The project supported by National Natural Science Foundation of China(81603587,81603668);Science and Technology Development Plan of Guangdong Province(2017A020211016);and Youth Medical Talent Fund of Guangzhou University of Chinese Medicine(QNYC20170105)

Yang CHEN,Tel:13500013336,E-mail：ychen8@gzucm.edu.cn