On the effects of NLRP3 inflammasome on mice pancreatic β-cell damage induced by advanced glycation end products
10.3760/cma.j.issn.1000-6699.2018.08.012
- VernacularTitle:NLRP3炎症小体在糖化终末产物诱导的小鼠胰岛β细胞损伤中的作用研究
- Author:
Xiang KONG
1
,
2
;
Qing SU
;
Hongmei ZHANG
;
Ning LIN
;
Xiaoyong LI
;
Zhen YANG
;
Ruyuan DENG
;
Chongxiao LIU
;
Jie JIN
;
Guangxun MENG
Author Information
1. 200092 上海交通大学医学院附属新华医院内分泌科
2. 241000 皖南医学院第一附属医院弋矶山医院内分泌科
- Keywords:
Advanced glycation end products;
NLRP3 inflammasome;
β-cell damage
- From:Chinese Journal of Endocrinology and Metabolism
2018;34(8):690-695
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the role of the pyrin domain-containing 3 ( NLRP3) inflammasome in advanced glycation end products ( AGEs )-induced mice pancreatic β-cell damage. Methods AGEs were administered intraperitoneally for 6 weeks in NLRP3 knockout mice or C57BL/6J mice. Intraperitoneal glucose tolerance test and insulin releasing test were performed. Pancreatic sections were stained with haematoxylin and eosin, or with F4/80 and NLRP3 antibodies. Insulin and pancreatic tissue monocyte chemotactic protein 1 ( MCP-1) as well as interleukin-1β( IL-1β) levels were measured with ELISA kits. Expression of MCP-1 protein was determined by western blot. MIN6 cells and mouse peritoneal macrophages cells were treated with AGEs and different interventions (antioxidant NAC, adenovirus NLRP3 shRNA or NLRP3 knockout). Reactive oxygen species production, NLRP3 mRNA expression, IL-1β secretion, caspase 1 activity, apoptosis and glucose stimulated insulin release were determined. Results Injection of AGEs induced an abnormal response to glucose, enhanced the insulitis score, and increased the levels of pancreatic tissue MCP-1 and IL-1β, as well as raised the expression of NLRP3 and F4/80 in pancreatic islet. Remarkably, co-localization of NLRP3 and macrophage marker F4/80 was observed in islet. The damages were improved in NLRP3 knockout mice. After incubation with AGEs, reactive oxygen species production and cell apoptosis was enhanced, NLRP3 inflammasome activated, with glucose-stimulated insulin release impaired in MIN6 cells. NAC treatment alliviated the above damages, but NLRP3 gene silencing had no effect on ROS level, apoptosis, and insulin secretion. Finally NAC treatment and NLRP3 gene knockout inhibited activation of NLRP3 inflammasome induced by AGEs in mouse peritoneal macrophages cells. Conclusion NLRP3 knockout ameliorates the islet β-cell damage induced by AGEs. These effects were associated with AGEs-induced islets macrophage infiltrating by up-regulation of MCP-1 expression, and AGEs-induced activation of NLRP3 inflammasome in macrophage through ROS pathway, which results in the release of active IL-1βand leads to the lesions of β-cell.
