CysLT2 receptor mediates lipopolysaccharide-induced microglial inflammation and consequent neurotoxicity in vitro.
Neuroinflammation induced by microglial activation plays a critical role in many neurodegenerative diseases, including Parkinson's disease (PD). Recent studies have indicated that cysteinyl leukotriene receptor 2 (CysLT2R) is involved in inflammation and brain injury after cerebral ischemia. However, the role of CysLT2R in microglial responses associated with PD remains unclear. In the present study, we determined the regulatory roles of CysLT2R in microglial inflammation and subsequent neurotoxicity in an in vitro brain inflammation model induced by the microglial activator lipopolysaccharide (LPS). We found that LPS induced phagocytosis of a murine microglial cell line (BV-2 cells) and increased production of the proinflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β). The expression of CysLT2R protein was up-regulated and the nuclear translocation of CysLT2R was induced in LPS-activated BV-2 cells. CysLT2R selective antagonist HAMI 3379 significantly inhibited LPS-induced phagocytosis and overproduction of the cytokines in BV-2 cells. Similarly, the CysLT2R silencing by specific short hairpin RNA (shRNA) had the same effects as those of HAMI 3379, suggesting that the effect might be CysLT2R-dependent. Furthermore, the conditioned medium (CM) derived from LPS-treated BV-2 cells induced the cell death of a rat adrenal pheochromocytoma cell line (PC12). HAMI 3379 and CysLT2R shRNA attenuated neuronal death by suppressing the production of neurotoxic cytokines released from LPS-activated microglia. Collectively, these results suggest that CysLT2R mediates LPS-induced microglial inflammation and consequent neurotoxicity. CysLT2R may be a promising molecular target that modulates microglia-related neuroinflammation in neurodegenerative disorders, such as PD.
Department of Pharmacology, Hangzhou Key Laboratory of Medical Neurobiology, School of Medicine, Hangzhou Normal University, Hangzhou 310036, PR China; Changzhou Hygiene Vocational Technology College, Changzhou 213002, PR China. Electronic address: chenlu19891128@163.com.
Lu
L
Chen
Department of Pharmacology, Hangzhou Key Laboratory of Medical Neurobiology, School of Medicine, Hangzhou Normal University, Hangzhou 310036, PR China. Electronic address: yyang@hznu.edu.cn.
Yi
Y
Yang
Department of Pharmacology, Hangzhou Key Laboratory of Medical Neurobiology, School of Medicine, Hangzhou Normal University, Hangzhou 310036, PR China. Electronic address: lct@hznu.edu.cn.
Chen-Tan
CT
Li
Department of Pharmacology, School of Medicine, Zhejiang University, Hangzhou 310058, PR China; Department of Clinical Medicine, School of Medicine, Zhejiang University, Hangzhou 310058, PR China. Electronic address: 3100102379@zju.edu.cn.
Si-Ran
SR
Zhang
Department of Pharmacology, Hangzhou Key Laboratory of Medical Neurobiology, School of Medicine, Hangzhou Normal University, Hangzhou 310036, PR China. Electronic address: Wei.zheng@hznu.edu.cn.
Wei
W
Zheng
Department of Pharmacology, School of Medicine, Zhejiang University, Hangzhou 310058, PR China. Electronic address: weieq2006@zju.edu.cn.
Er-Qing
EQ
Wei
Department of Pharmacology, Hangzhou Key Laboratory of Medical Neurobiology, School of Medicine, Hangzhou Normal University, Hangzhou 310036, PR China. Electronic address: lhzhang@hznu.edu.cn.
Li-Hui
LH
Zhang
D000818:Animals; D016923:Cell Death / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002461:Cell Line, Transformed / Q000187:drug effects; D003509:Cyclohexanecarboxylic Acids / Q000494:pharmacology; D016207:Cytokines / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects*; D008070:Lipopolysaccharides / Q000633:toxicity*; D051379:Mice; D017628:Microglia / Q000187:drug effects*; D010587:Phagocytosis / Q000187:drug effects; D010795:Phthalic Acids / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000494:pharmacology; D051381:Rats; D018077:Receptors, Leukotriene / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors
C549635:3-(((3-carboxycyclohexyl)amino)carbonyl)-4-(3-(4-(4-(cyclohexyloxy)butoxy)phenyl)propoxy)benzoic acid; D003509:Cyclohexanecarboxylic Acids; D016207:Cytokines; D008070:Lipopolysaccharides; D010795:Phthalic Acids; D034741:RNA, Small Interfering; D018077:Receptors, Leukotriene; C412375:cysteinyl leukotriene receptor 2
Antagonist; Cysteinyl leukotriene receptor 2; Inflammation; Microglia; Neurotoxicity; RNA interference