title: Sinomenine activates astrocytic dopamine D2 receptors and alleviates neuroinflammatory injury via the CRYAB/STAT3 pathway after ischemic stroke in mice.
abstract: Astrocyte-mediated neuroinflammation plays a critical role in ischemic stroke-induced secondary cerebral injury. Previous studies have suggested that the dopamine D2 receptor (DRD2) acts as a key target in regulating the neuroinflammatory response. However, the underlying molecular mechanisms are still unknown, and effective DRD2 agonists are lacking. In the present study, we examined the anti-inflammatory and neuroprotective effects of sinomenine (Sino), a monomeric compound with potential immunoregulatory properties in nervous system.
  
  
  
  TTC staining, apoptosis assay, evaluation of brain edema, and neurological assessment were performed in the middle cerebral artery occlusion (MCAO) mouse model. Primary astrocytes exposed to oxygen glucose deprivation (OGD) were used in the in vitro experiments. Quantitative PCR was applied to assess the levels of inflammatory cytokines. Multi-labeling immunofluorescence, Western blot, co-immunoprecipitation, and electrophoretic mobility shift assay (EMSA) were also used to investigate the molecular mechanisms underlying the Sino-mediated anti-inflammatory effects in vivo and in vitro.
  
  
  
  Sino remarkably attenuated the cerebral infarction and neuronal apoptosis, reduced the levels of inflammatory cytokines, and alleviated neurological deficiency in MCAO mice. Sino significantly inhibited astrocytic activation and STAT3 phosphorylation as well as increased DRD2 and αB-crystallin (CRYAB) expression after MCAO. In vitro, Sino blocked OGD-induced activation of STAT3 and generation of pro-inflammatory cytokines in primary astrocytes, and these effects were significantly abolished by either DRD2 or CRYAB knockdown. Additionally, Sino induced up-regulation and nuclear translocation of CRYAB in astrocytes and enhanced the interaction between CRYAB and STAT3, which further inhibited the activation and DNA-binding activity of STAT3.
  
  
  
  Our study demonstrates that Sino activates astrocytic DRD2 and thereby suppresses neuroinflammation via the CRYAB/STAT3 pathway, which sheds some light on a promising therapeutic strategy for ischemic stroke.
authors.affiliation: Department of Neurology, The General Hospital of Shenyang Military Region, Shenyang, Liaoning, 110016, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Qiu
authors.affiliation: Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, Shaanxi, 710038, People's Republic of China.
authors.forename: Zhongjun
authors.initials: Z
authors.lastname: Yan
authors.affiliation: Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, Shaanxi, 710038, People's Republic of China.
authors.forename: Kai
authors.initials: K
authors.lastname: Tao
authors.affiliation: Department of Neurology, The 463rd Hospital of PLA, Shenyang, Liaoning, 110042, People's Republic of China.
authors.forename: Yansong
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, Shaanxi, 710038, People's Republic of China.
authors.forename: Yuqian
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Neurosurgery, The General Hospital of Shenyang Military Region, Shenyang, Liaoning, 110016, People's Republic of China.
authors.forename: Jingchen
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Neurosurgery, The General Hospital of Shenyang Military Region, Shenyang, Liaoning, 110016, People's Republic of China.
authors.forename: Yushu
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, Shaanxi, 710038, People's Republic of China. dayunfmmu@163.com.
authors.forename: Dayun
authors.initials: D
authors.lastname: Feng
authors.affiliation: Department of Neurology, The General Hospital of Shenyang Military Region, Shenyang, Liaoning, 110016, People's Republic of China. chsshenyang@sina.cn.
authors.forename: Huisheng
authors.initials: H
authors.lastname: Chen
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D018501:Antirheumatic Agents / Q000494:pharmacology / Q000627:therapeutic use; D001253:Astrocytes / Q000187:drug effects*; D001929:Brain Edema / Q000188:drug therapy / Q000209:etiology; D002478:Cells, Cultured; D002544:Cerebral Infarction / Q000188:drug therapy / Q000209:etiology; D004195:Disease Models, Animal; D004660:Encephalitis / Q000188:drug therapy / Q000209:etiology / Q000473:pathology; D000860:Hypoxia / Q000188:drug therapy; D020244:Infarction, Middle Cerebral Artery / Q000150:complications*; D051379:Mice; D008810:Mice, Inbred C57BL; D009019:Morphinans / Q000494:pharmacology* / Q000627:therapeutic use*; D009422:Nervous System Diseases / Q000188:drug therapy / Q000209:etiology; D034622:RNA Interference / Q000502:physiology; D017448:Receptors, Dopamine D2 / Q000378:metabolism*; D050796:STAT3 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D038203:alpha-Crystallin B Chain / Q000378:metabolism
chemical_list: D018501:Antirheumatic Agents; C525420:Cryab protein, mouse; D009019:Morphinans; D017448:Receptors, Dopamine D2; D050796:STAT3 Transcription Factor; D038203:alpha-Crystallin B Chain; C009271:sinomenine
keywords: Astrocyte; Dopamine D2 receptor (DRD2); Ischemic stroke; Neuroinflammation; Sinomenine