title: Age-related increases in amyloid beta and membrane attack complex: evidence of inflammasome activation in the rodent eye.
abstract: BACKGROUND
  The membrane attack complex (MAC) is a key player in the pathogenesis of age-related macular degeneration (AMD) and is a putative activator of the NLRP3 inflammasome. Amyloid beta (Aβ), a component of drusen deposits, has also been implicated in inflammasome activation by our work and those of others. However, the interactions of MAC and Aβ are still poorly understood, especially their roles in aging and retinal degenerative pathologies. Since inflammasome activation may represent a key cellular pathway underlying age-related chronic inflammation in the eye, the purpose of this study is to identify the effects associated with MAC and inflammasome activation in the retinal pigment epithelium (RPE)/choroid and to evaluate the therapeutic merits of MAC suppression.
  
  
  METHODS
  Adult Long-Evans rats were divided into treatment and control groups. Treatment groups received oral aurin tricarboxylic acid complex (ATAC), a MAC inhibitor, in drinking-water, and control groups received drinking-water alone (No ATAC). Groups were sacrificed at 7.5 or 11.5 months, after approximately 40 days of ATAC treatment. To study age-related changes of Aβ and MAC in RPE/choroid, naive animals were sacrificed at 2.5, 7.5, and 11.5 months. Eye tissues underwent immunohistochemistry and western blot analysis for MAC, Aβ, NF-κB activation, as well as cleaved caspase-1 and IL-18. Vitreal samples were collected and assessed by multiplex assays for secreted levels of IL-18 and IL-1β. Statistical analyses were performed, and significance level was set at p ≤ 0.05.
  
  
  RESULTS
  In vivo studies demonstrated an age-dependent increase in MAC, Aβ, and NF-κB activation in the RPE/choroid. Systemic ATAC resulted in a prominent reduction in MAC formation and a concomitant reduction in inflammasome activation measured by cleaved caspase-1 and secreted levels of IL-18 and IL-1β, but not in NF-κB activation. In vitro studies demonstrated Aβ-induced MAC formation on RPE cells.
  
  
  CONCLUSIONS
  Age-dependent increases in Aβ and MAC are present in the rodent outer retina. Our results suggest that suppressing MAC formation and subsequent inflammasome activation in the RPE/choroid may reduce chronic low-grade inflammation associated with IL-18 and IL-1β in the outer retina.
authors.affiliation: Department of Ophthalmology and Visual Sciences, Faculty of Medicine, University of British Columbia, 2550 Willow Street, Vancouver, V5Z 3N9, BC, Canada. sixi@ualberta.ca.
authors.forename: Tom
authors.initials: T
authors.lastname: Zhao
authors.affiliation: Department of Ophthalmology and Visual Sciences, Faculty of Medicine, University of British Columbia, 2550 Willow Street, Vancouver, V5Z 3N9, BC, Canada. wade.jygao@alumni.ubc.ca.
authors.forename: Jiangyuan
authors.initials: J
authors.lastname: Gao
authors.affiliation: Department of Ophthalmology and Visual Sciences, Faculty of Medicine, University of British Columbia, 2550 Willow Street, Vancouver, V5Z 3N9, BC, Canada. jeniferv@pacificu.edu.
authors.forename: Jenifer
authors.initials: J
authors.lastname: Van
authors.affiliation: Department of Ophthalmology and Visual Sciences, Faculty of Medicine, University of British Columbia, 2550 Willow Street, Vancouver, V5Z 3N9, BC, Canada. ect@mail.ubc.ca.
authors.forename: Eleanor
authors.initials: E
authors.lastname: To
authors.affiliation: Department of Ophthalmology and Visual Sciences, Faculty of Medicine, University of British Columbia, 2550 Willow Street, Vancouver, V5Z 3N9, BC, Canada. wangaikun@hotmail.com.
authors.forename: Aikun
authors.initials: A
authors.lastname: Wang
authors.affiliation: Department of Ophthalmology and Visual Sciences, Faculty of Medicine, University of British Columbia, 2550 Willow Street, Vancouver, V5Z 3N9, BC, Canada. sijia.cao@alumni.ubc.ca.
authors.forename: Sijia
authors.initials: S
authors.lastname: Cao
authors.affiliation: Department of Ophthalmology and Visual Sciences, Faculty of Medicine, University of British Columbia, 2550 Willow Street, Vancouver, V5Z 3N9, BC, Canada. jcui@mail.ubc.ca.
authors.forename: Jing Z
authors.initials: JZ
authors.lastname: Cui
authors.affiliation: Kinsmen Lab of Neurological Research, University of British Columbia, Vancouver, BC, Canada. jjguo@mail.ubc.ca.
authors.forename: Jian-Ping
authors.initials: JP
authors.lastname: Guo
authors.affiliation: Kinsmen Lab of Neurological Research, University of British Columbia, Vancouver, BC, Canada. moonhee.lee@ubc.ca.
authors.forename: Moonhee
authors.initials: M
authors.lastname: Lee
authors.affiliation: Kinsmen Lab of Neurological Research, University of British Columbia, Vancouver, BC, Canada. mcgeerpl@mail.ubc.ca.
authors.forename: Patrick L
authors.initials: PL
authors.lastname: McGeer
authors.affiliation: Department of Ophthalmology and Visual Sciences, Faculty of Medicine, University of British Columbia, 2550 Willow Street, Vancouver, V5Z 3N9, BC, Canada. jms@mail.ubc.ca.
authors.forename: Joanne A
authors.initials: JA
authors.lastname: Matsubara
mesh_terms: D000375:Aging / Q000378:metabolism*; D016229:Amyloid beta-Peptides / Q000378:metabolism*; D000818:Animals; D001312:Aurintricarboxylic Acid / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism*; D002829:Choroid / Q000187:drug effects / Q000378:metabolism*; D015938:Complement Membrane Attack Complex / Q000378:metabolism*; D004195:Disease Models, Animal; D058847:Inflammasomes / Q000378:metabolism*; D020382:Interleukin-18 / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism; D008268:Macular Degeneration / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D051381:Rats; D020318:Rats, Long-Evans; D012160:Retina / Q000187:drug effects / Q000378:metabolism*
chemical_list: D016229:Amyloid beta-Peptides; D002352:Carrier Proteins; D015938:Complement Membrane Attack Complex; D058847:Inflammasomes; D020382:Interleukin-18; D053583:Interleukin-1beta; D016328:NF-kappa B; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C573346:Nlrp3 protein, rat; D001312:Aurintricarboxylic Acid