Activation of Platelet NLRP3 Inflammasome in Crohn's Disease

Ge Zhang; He Chen; Yifan Guo; Wei Zhang; Qiuyu Jiang; Si Zhang; Liping Han; She Chen; Ruyi Xue

doi:10.3389/fphar.2021.705325

Activation of Platelet NLRP3 Inflammasome in Crohn's Disease

Front Pharmacol. 2021 Jun 28:12:705325. doi: 10.3389/fphar.2021.705325. eCollection 2021.

Authors

Ge Zhang¹, He Chen², Yifan Guo³, Wei Zhang¹, Qiuyu Jiang⁴, Si Zhang¹, Liping Han¹, She Chen¹, Ruyi Xue²

Affiliations

¹ NHC Key Laboratory of Glycoconjugates Research, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China.
² Department of Gastroenterology and Hepatology, Shanghai Institute of Liver Diseases, Zhongshan Hospital, Fudan University, Shanghai, China.
³ Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, China.
⁴ Shanghai Medical College, Fudan University, Shanghai, China.

Abstract

Patients with Crohn's disease (CD) are inclined to have platelet hyperactivity and an increased risk of intestinal micro-thrombosis. However, the mechanisms underlying platelet hyperactivity in CD are not well understood. We investigated the assembly of platelet NLRP3 inflammasome in patients with active CD and its correlation with platelet hyperactivity. In this study, Real-time PCR and western blotting analyses uncovered that ASC, NLRP3, and active caspase-1 were significantly upregulated in platelets from patients with active CD compared with healthy subjects. As revealed by flow cytometry (FCM) and ELISA analyses, the levels of interleukin-1β in both serum and isolated platelets were elevated in patients with active CD. Co-immunoprecipitation and immunofluorescence experiments revealed an increased assembly of NLRP3 inflammasome in platelets from patients with active CD. In addition, higher levels of intracellular reactive oxygen species (ROS) were observed in these platelets by FCM. Furthermore, elevated levels of platelet P-selectin exposure and fibrinogen binding were demonstrated in patients with active CD by FCM. They were positively correlated with the protein levels of NLRP3 inflammasome components. Collectively, our results indicate that the ROS-NLRP3 inflammasome-interleukin-1β axis may contribute to platelet hyperactivity in active CD.

Keywords: Crohn’s disease; NLRP3 inflammasome; ROS; interleukin-1β; platelet hyperactivity.