title: Characterization of a Synthetic Steroid 24-keto-cholest-5-en-3β, 19-diol as a Neuroprotectant.
abstract: BACKGROUND
  Neuroactive steroids represent promising candidates for the treatment of neurological disorders. Our previous studies identified an endogenous steroid cholestane-3β, 5α, 6β-triol (Triol) as a novel neuroprotectant.
  
  
  OBJECTIVE
  We aimed to identify a potent candidate for stroke treatment through a screening of Triol analogs.
  
  
  METHODS
  Hypoxia- and glutamate-induced neuronal injury models in vitro, middle cerebral artery occlusion (MCAO)-induced cerebral ischemia model in vivo, fluorescein diacetate (FDA) for alive and propidium iodide (PI) for dead staining, LDH assay, and calcium imaging techniques were used.
  
  
  RESULTS
  24-keto-cholest-5-en-3β, 19-diol (Diol) showed the most potent neuroprotective effect among the screened structurally related compounds. FDA and PI staining showed that Diol concentration dependently increased the survival rate of cerebellar granule neurons (CGNs) challenged with glutamate or hypoxia, with an effective threshold concentration of 2.5 μM. Consistently, the quantitative LDH release assay showed the same concentration-dependent protection in both models. Diol, at 10 μM, potently decreased glutamate- and hypoxia-induced LDH release from 51.6 to 18.2% and 62.1 to 21.7%, respectively, which values are close to the normal LDH release (~16-18%). Moreover, we found Diol effectively decreased MCAO-induced infarction volume in mice from ~23% to 7%, at a dose of 6 mg/kg. We further explored the underlying mechanism and found that Diol attenuated NMDA-induced intracellular calcium ([Ca(2+) ]i ) increase in cortical neurons, suggesting a negative modulatory effect on NMDA receptor.
  
  
  CONCLUSIONS
  Taken together, we identified Diol as a potent neuroprotectant. It may represent a novel and promising neuroprotectant for stroke intervention.
authors: Department of Pharmacology, Zhongshan Medical College, Sun Yat-Sen University, Guangzhou, China.
authors: Min
authors: M
authors: Yan
authors: Department of Pharmacology, Zhongshan Medical College, Sun Yat-Sen University, Guangzhou, China.
authors: Ai-Ling
authors: AL
authors: Liu
authors: School of pharmaceutical sciences, Sun Yat-Sen University, Guangzhou, China.
authors: Shu-Jia
authors: SJ
authors: Zhou
authors: Department of Pharmacology, Zhongshan Medical College, Sun Yat-Sen University, Guangzhou, China.
authors: Li-Peng
authors: LP
authors: Tang
authors: Department of Cardiovascular Epidemiology, Guangdong General Hospital, Guangzhou, China.
authors: Yan-Qiu
authors: YQ
authors: Ou
authors: Department of Pharmacology, Zhongshan Medical College, Sun Yat-Sen University, Guangzhou, China.
authors: Wei
authors: W
authors: Yin
authors: School of pharmaceutical sciences, Sun Yat-Sen University, Guangzhou, China.
authors: Xin-Ying
authors: XY
authors: Chen
authors: Department of Pharmacology, Zhongshan Medical College, Sun Yat-Sen University, Guangzhou, China.
authors: Xing-Wen
authors: XW
authors: Su
authors: Department of Pharmacology, Zhongshan Medical College, Sun Yat-Sen University, Guangzhou, China.
authors: Peng-Xin
authors: PX
authors: Qiu
authors: Department of Pharmacology, Zhongshan Medical College, Sun Yat-Sen University, Guangzhou, China.
authors: Yi-Jun
authors: YJ
authors: Huang
authors: School of pharmaceutical sciences, Sun Yat-Sen University, Guangzhou, China.
authors: Jing-Xia
authors: JX
authors: Zhang
authors: Department of Pharmacology, Zhongshan Medical College, Sun Yat-Sen University, Guangzhou, China.
authors: Guang-Mei
authors: GM
authors: Yan
authors: Department of Pharmacology, Zhongshan Medical College, Sun Yat-Sen University, Guangzhou, China.
authors: Tian-Dong
authors: TD
authors: Leng
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002118:Calcium / Q000378:metabolism; D015687:Cell Hypoxia / Q000187:drug effects; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology; D002777:Cholestanols / Q000737:chemistry / Q000627:therapeutic use*; D004195:Disease Models, Animal; D018690:Excitatory Amino Acid Agonists / Q000633:toxicity; D018698:Glutamic Acid / Q000633:toxicity; D020244:Infarction, Middle Cerebral Artery / Q000188:drug therapy*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D016202:N-Methylaspartate / Q000633:toxicity; D009474:Neurons / Q000187:drug effects*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D010100:Oxygen / Q000008:administration & dosage; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D002777:Cholestanols; D018690:Excitatory Amino Acid Agonists; D018696:Neuroprotective Agents; C000474:cholestane-3,5,6-triol; D018698:Glutamic Acid; D016202:N-Methylaspartate; D007770:L-Lactate Dehydrogenase; D010100:Oxygen; D002118:Calcium
keywords: NMDA; Neuroprotection; Steroid; Stroke