[Abstact]Objective To investigate the protective effects of diosmin(DSM)on human neuroblastoma SH-SY5Y cells treated with oxygen-glucose deprivation/reoxygenation(OGD/R)via mitochondrial pathway.Methods SH-SY5Y cells were cultured in vitro,OGD/R model was established,and the optimal concentration of DSM was determined by CCK-8 assay and subsequent experiments were conducted.The cells were divided into blank control group(Control group),OGD/R group,and DSM group(DSM concentration was 0.01,0.02,and 0.04 g/L,respectively).Cell survival rate was detected by CCK-8 assay,cell morphology was observed by phase-contrast microscope,intracellular ATP levels and mitochondrial complex Ⅱ activity were measured by spectrophotometry,mitochondrial membrane potential was detected by JC-1 staining method,and apoptosis was detected via TUNEL assay.Results DSM at three different concentrations(0.01,0.02,and 0.04 g/L)promoted the survival of SH-SY5Y cells damaged by OGD/R,with 0.02 g/L concentration exhibiting the most significant effect on cell survival.Compared with Control group,OGD/R group was characterized by shrunken cell morphology,significantly decreased ATP content and mitochondrial complex Ⅱ activity,decreased mitochondrial membrane potential,and increased apoptosis rate.After pre-incubation with 0.02 g/L DSM for 3 h,the survival rate of OGD/R damaged cells was increased,cell morphology was basically normal,ATP content and mitochondrial complex Ⅱ activity were significantly increased,mitochondrial membrane potential was significantly increased,and cell apoptosis was decreased.Conclusion DSM has a significant protective effect on SH-SY5Y cells damaged by OGD/R and the best protective effect is displayed at 0.02 g/L,and its mechanism of action is closely related to maintaining the normal function of mitochondria and reducing the state of apoptosis.
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