Preventive effect of genetic knockdown and pharmacological blockade of CysLT1R on lipopolysaccharide (LPS)-induced memory deficit and neurotoxicity in vivo.
Previously we reported that cysteinyl leukotrienes (Cys-LTs) and the type 1 receptor for Cys-LTs (CysLT1R) are related to amyloid β (Aβ)-induced neurotoxicity. The aim of the current study was to find out the role of CysLT1R on lipopolysaccharide (LPS)-induced cognitive deficit and neurotoxicity. shRNA-mediated knockdown or pharmacological blockade (by pranlukast) of CysLT1R were performed in ICR mice for 21days prior to systemic infusion of LPS. From day 22, LPS was administered for 7days and then a set of behavioral, histopathological and biochemical tests were employed to test memory, neuroinflammation and apoptotic responses in the mouse hippocampus. LPS (only)-treated mice showed poor performance in both Morris water maze (MWM) and Y-maze tests. However, shRNA-mediated knockdown or pranlukast-treated blockade of CysLT1R improved performance of the mice in these tests. To find out the possible underlying mechanisms, we assessed several parameters such as microglial activation (by immunohistochemistry), level of CysLT1R (by WB and qRT-PCR) and the inflammatory/apoptotic pathways (by ELISA or TUNEL or WB) in the mouse hippocampus. LPS-induced memory impairment was accompanied by activation of microglia, higher level of CysLT1R, IL-1β, TNF-α and nuclear NF-κB p65. LPS also caused apoptosis in the hippocampus as detected by TUNEL staining, further supplemented by detection of increased Caspase-3 and a reduced Bcl-2/Bax ratio. All of these adverse changes in the mouse hippocampus were inhibited by pretreatment with CysLT1R-shRNA and pranlukast. Through this study we suggest that CysLT1R shares a strong correlation with LPS-associated memory deficit, neuroinflammation and apoptosis and CysLT1R could be a novel target for preventive measures to intervene the progression of Alzheimer's disease (AD)-like phenotypes.
Department of Pharmacology, Jiangsu Key Laboratory of Drug Discovery for Metabolic Diseases, and State Key Laboratory for Natural Medicines, China Pharmaceutical University, Nanjing 210009, Jiangsu, China.
Fang
F
Chen
Department of Pharmacology, Jiangsu Key Laboratory of Drug Discovery for Metabolic Diseases, and State Key Laboratory for Natural Medicines, China Pharmaceutical University, Nanjing 210009, Jiangsu, China.
Arijit
A
Ghosh
School of Pharmacy, Nantong University, Nantong 226001, Jiangsu, China.
Feng
F
Wu
Department of Pharmacology, Jiangsu Key Laboratory of Drug Discovery for Metabolic Diseases, and State Key Laboratory for Natural Medicines, China Pharmaceutical University, Nanjing 210009, Jiangsu, China.
Susu
S
Tang
Department of Pharmacology, Jiangsu Key Laboratory of Drug Discovery for Metabolic Diseases, and State Key Laboratory for Natural Medicines, China Pharmaceutical University, Nanjing 210009, Jiangsu, China.
Mei
M
Hu
Department of Pharmacology, Jiangsu Key Laboratory of Drug Discovery for Metabolic Diseases, and State Key Laboratory for Natural Medicines, China Pharmaceutical University, Nanjing 210009, Jiangsu, China.
Hongbin
H
Sun
Department of Pharmacology, Jiangsu Key Laboratory of Drug Discovery for Metabolic Diseases, and State Key Laboratory for Natural Medicines, China Pharmaceutical University, Nanjing 210009, Jiangsu, China.
Lingyi
L
Kong
Department of Pharmacology, Jiangsu Key Laboratory of Drug Discovery for Metabolic Diseases, and State Key Laboratory for Natural Medicines, China Pharmaceutical University, Nanjing 210009, Jiangsu, China. Electronic address: honghao@cpu.edu.cn.
Hao
H
Hong
D016229:Amyloid beta-Peptides / Q000378:metabolism; D000818:Animals; D003072:Cognition Disorders / Q000235:genetics / Q000378:metabolism; D060825:Cognitive Dysfunction / Q000378:metabolism; D055785:Gene Knockdown Techniques / Q000379:methods; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008297:Male; D008568:Memory / Q000187:drug effects; D008569:Memory Disorders / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D020258:Neurotoxicity Syndromes / Q000235:genetics / Q000378:metabolism; D018077:Receptors, Leukotriene / Q000235:genetics*
D016229:Amyloid beta-Peptides; D008070:Lipopolysaccharides; D018077:Receptors, Leukotriene; C083169:leukotriene D4 receptor
Alzheimer’s disease; Bcl-2/Bax; Caspase-3; CysLT(1)R; IL-1β; Microglia; NF-κB p65; Pranlukast; TNF-α; shRNA