title: SMND-309 promotes neuron survival through the activation of the PI3K/Akt/CREB-signalling pathway.
abstract: Context In clinical practice, the promotion of neuron survival is necessary to recover neurological functions after the onset of stroke. Objective This study aimed to investigate the post-ischaemic neuroprotective effect of SMND-309, a novel metabolite of salvianolic acid, on differentiated SH-SY5Y cells. Materials and methods SH-SY5Y cells were differentiated by pre-treating with 5 μM all-trans-retinoic acid for 6 d. The differentiated SH-SY5Y cells were exposed to oxygen-glucose deprivation (OGD) for 2 h and reperfusion (R) for 24 h to induce OGD/R injury. After OGD injury, differentiated SH-SY5Y cells were treated with or without SMND-309 (5, 10, 20 μM) for another 24 h. Cell viability was detected through Cell counting kit-8 assay and lactate dehydrogenase leakage assay. Apoptosis was evaluated through flow cytometry, caspase-3 activity assay. Changes in protein levels were assessed through Western blot. Results SMND-309 ameliorated the degree of injury in the differentiated SH-SY5Y cells by increasing cell viabilities (5 μM, 65.4% ± 4.1%; 10 μM, 69.8% ± 3.7%; 20 μM, 75.3% ± 5.1%) and by reducing LDH activity (20 μM, 2.5 fold) upon OGD/R stimulation. Annexin V-fluorescein isothiocyanate/propidium iodide staining results suggested that apoptotic rate of differentiated SH-SY5Y cells decreased from 43.8% induced by OGD/R injury to 19.2% when the cells were treated with 20 μM SMND-309. SMND-309 significantly increased the Bcl-2 level of the injured differentiated SH-SY5Y cells but decreased the caspase-3 activity of these cells by 1.6-fold. In contrast, SMND-309 did not affect the Bax level of these cells. SMND-309 evidently increased the protein expression of BDNF when Akt and CREB were activated. This function was antagonized by the addition of LY294002. Conclusion SMND-309 can prevent neuronal cell death in vitro. This process may be related to the activation of the PI3K/Akt/CREB-signalling pathway.
authors.affiliation: a School of Special Education , Binzhou Medical University , Yantai , PR China ;
authors.forename: Youlei
authors.initials: Y
authors.lastname: Wang
authors.affiliation: a School of Special Education , Binzhou Medical University , Yantai , PR China ;
authors.forename: Jinjin
authors.initials: J
authors.lastname: Zhang
authors.affiliation: b Zibo Occupational Disease Hospital , Zibo , PR China ;
authors.forename: Meng
authors.initials: M
authors.lastname: Han
authors.affiliation: a School of Special Education , Binzhou Medical University , Yantai , PR China ;
authors.forename: Bo
authors.initials: B
authors.lastname: Liu
authors.affiliation: a School of Special Education , Binzhou Medical University , Yantai , PR China ;
authors.forename: Yulin
authors.initials: Y
authors.lastname: Gao
authors.affiliation: a School of Special Education , Binzhou Medical University , Yantai , PR China ;
authors.forename: Peng
authors.initials: P
authors.lastname: Ma
authors.affiliation: c School of Pharmacy , Taishan Medical College , Taian , PR China ;
authors.forename: Songzi
authors.initials: S
authors.lastname: Zhang
authors.affiliation: a School of Special Education , Binzhou Medical University , Yantai , PR China ;|d Department of Otolaryngology - HNS , Case Western Reserve University , Cleveland , OH , USA.
authors.forename: Qingyin
authors.initials: Q
authors.lastname: Zheng
authors.affiliation: a School of Special Education , Binzhou Medical University , Yantai , PR China ;
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Song
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D002109:Caffeic Acids / Q000494:pharmacology*; D015687:Cell Hypoxia; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005947:Glucose / Q000172:deficiency; D006801:Humans; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D018696:Neuroprotective Agents / Q000494:pharmacology*; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: C539487:2-(6-(2-carboxyvinyl)-2,3-dihydroxyphenyl)-3-(3,4-dihydroxyphenyl)acrylic acid; D051017:Apoptosis Regulatory Proteins; C115163:CREB1 protein, human; D002109:Caffeic Acids; D017362:Cyclic AMP Response Element-Binding Protein; D018696:Neuroprotective Agents; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors; D058539:Phosphatidylinositol 3-Kinase; D051057:Proto-Oncogene Proteins c-akt; D005947:Glucose
keywords: Apoptosis; differentiated SH-SY5Y cells; oxygen-glucose deprivation/reperfusion