title: NLRP3 is Required for Complement-Mediated Caspase-1 and IL-1beta Activation in ICH.
abstract: Complement-mediated inflammation plays a vital role in intracerebral hemorrhage (ICH), implicating pro-inflammatory factor interleukin-1beta (IL-1β) secretion. Brain samples and contralateral hemiencephalon were all collected and detected by Western blot. NLRP3 expression was located by dual immunofluorescence staining at 1, 3, and 5 days post-ICH. Brain water content was examined post-ICH. The neural deficit scores were evaluated by observers blindly. ILs were detected by ELISA. SiRNAs targeting NLRP3 (siNLRP3), siASC, and siControl were injected to inhibit NLRP3 function. To test the complement activation via Nod-like receptor (NLR) family pyrin domain-containing 3 (NLRP3), normal rabbit complement (NRC) was injected with lipopolysaccharide (LPS) to facilitate the complement function. As a result, complement 3a (C3a) and complement 5a (C5a) were upregulated during the ICH-induced neuroinflammation, and ablation of C3 attenuates ICH-induced IL-1β release. Though the LPS rescues the neuroinflammation in the ICH model, C3 deficiency attenuates the LPS-induced inflammatory effect. The NLRP3 inflammasome was activated after ICH and was located in the microglial cell of the mouse brain, which exhibits a time-dependent manner. However, the number of NLRP3/Iba-1 dual-labeled cells in the C3-/-group is less than that in the WT group in each time course, respectively. IL-1β and IL-18 released in perihematoma tissue, caspase-1-p20, brain water content, and behavioral outcomes were attenuated in the siNLRP3 and siASC groups than in the siControl and ICH groups. We also found that 5% of complement supplement enhances ICH-induced IL-1β release, while NLRP3 and ASC inhibition attenuates it. In conclusion, complement-induced ICH neuroinflammation depended on NLRP3 activation, which facilities LPS- and ICH-induced neuroinflammation, and NLRP3 is required for ICH-induced inflammation.
authors.affiliation: Department of Cerebrovascular Disease, The First Affiliated Hospital of Zunyi Medical College, No. 139, Dalian Avenue, Huichuan District, Zunyi, Guizhou, 563000, China.
authors.forename: Sheng-Tao
authors.initials: ST
authors.lastname: Yao
authors.affiliation: Department of Cerebrovascular Disease, The First Affiliated Hospital of Zunyi Medical College, No. 139, Dalian Avenue, Huichuan District, Zunyi, Guizhou, 563000, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Cao
authors.affiliation: Department of Neonatal, The Third Affiliated Hospital of Zunyi Medical College, No. 98, Phoenix Rd, Zunyi, Guizhou, 563002, China.
authors.forename: Jia-Lin
authors.initials: JL
authors.lastname: Chen
authors.affiliation: Department of Cerebrovascular Disease, The First Affiliated Hospital of Zunyi Medical College, No. 139, Dalian Avenue, Huichuan District, Zunyi, Guizhou, 563000, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.affiliation: Department of Cerebrovascular Disease, The First Affiliated Hospital of Zunyi Medical College, No. 139, Dalian Avenue, Huichuan District, Zunyi, Guizhou, 563000, China.
authors.forename: Rui-Ming
authors.initials: RM
authors.lastname: Fan
authors.affiliation: Department of Cerebrovascular Disease, The First Affiliated Hospital of Zunyi Medical College, No. 139, Dalian Avenue, Huichuan District, Zunyi, Guizhou, 563000, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Li
authors.affiliation: Department of Cerebrovascular Disease, The First Affiliated Hospital of Zunyi Medical College, No. 139, Dalian Avenue, Huichuan District, Zunyi, Guizhou, 563000, China.
authors.forename: You-Chao
authors.initials: YC
authors.lastname: Zeng
authors.affiliation: Department of Cerebrovascular Disease, The First Affiliated Hospital of Zunyi Medical College, No. 139, Dalian Avenue, Huichuan District, Zunyi, Guizhou, 563000, China.
authors.forename: Song
authors.initials: S
authors.lastname: Jiao
authors.affiliation: Department of Cerebrovascular Disease, The First Affiliated Hospital of Zunyi Medical College, No. 139, Dalian Avenue, Huichuan District, Zunyi, Guizhou, 563000, China.
authors.forename: Xiang-Ping
authors.initials: XP
authors.lastname: Xia
authors.affiliation: Department of Cerebrovascular Disease, The First Affiliated Hospital of Zunyi Medical College, No. 139, Dalian Avenue, Huichuan District, Zunyi, Guizhou, 563000, China.
authors.forename: Chong
authors.initials: C
authors.lastname: Han
authors.affiliation: Department of Cerebrovascular Disease, The First Affiliated Hospital of Zunyi Medical College, No. 139, Dalian Avenue, Huichuan District, Zunyi, Guizhou, 563000, China. neurosci_neurology@163.com.
authors.forename: Qi-Shan
authors.initials: QS
authors.lastname: Ran
mesh_terms: D000818:Animals; D001921:Brain / Q000276:immunology; D020170:Caspase 1 / Q000378:metabolism*; D002543:Cerebral Hemorrhage / Q000276:immunology*; D015926:Complement C3a / Q000276:immunology; D015936:Complement C5a / Q000276:immunology; D058847:Inflammasomes / Q000276:immunology*; D053583:Interleukin-1beta / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D017628:Microglia / Q000276:immunology; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein / Q000378:metabolism*; D020078:Neurogenic Inflammation / Q000276:immunology
chemical_list: D058847:Inflammasomes; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C492716:Nlrp3 protein, mouse; D015926:Complement C3a; D015936:Complement C5a; D020170:Caspase 1
keywords: Complement, NLRP3 inflammasome; IL-1β; Intracerebral hemorrhage; Neuroinflammation