1.Effect of lycium barbarum polysaccharides of apoptosis and autophagic death in primary cultured hippocampal neurons injured by oxygen glucose deprivation and reoxygenation
Yang YU ; Xinghui LI ; Jingnan PU ; Xiuquan WU ; Peng LUO ; Jiu WANG ; Zhou FEI
Chinese Journal of Neuromedicine 2018;17(7):649-655
Objective To investigate the effect of lycium barbarum polysaccharides (LBP) on apoptosis and autophagic death in primary cultured hippocampal neurons after oxygen glucose deprivation and reoxygenation (OGD/R) injury.Methods Primary cultured hippocampal neurons were exposed to OGD/R.The cells were randomly divided into 5 groups:control group,OGD/R group,and OGD/R+LBP groups (15,30 and 60 μg/mL LBP).The cell viability was assessed by MTT assay.The cell damage was evaluated through detecting the lactate dehydrogenase (LDH) release rate.Apoptosis rate was detected by TUNEL,and cleaved Caspase-3 was identified by immunofluorescence staining.Protein expression was detected by Western blotting.Results As compared with the control group,OGD/R group had significantly decreased cell viability (P<0.05);and significantly increased cell viability and decreased LDH release rate were noted in the LBP (15,30 and 60 μg/mL) treatment groups as compared with those in the OGD/R group (P<0.05).The 60 μg/mL LBP treatment group had significantly smaller number of TUNEL-positive cells than the OGD/R group(P<0.05).Immunofluorescence staining and Western blotting both revealed that 60 μg/mL LBP treatment group had significantly decreased Beclinl level and ratio of cleaved Caspase-3/Caspase-3 and ratio of microtubule-associated protein light chain (LC)3 Ⅱ/LC3Ⅰ,and statistically increased p62 level and ratio of Bcl-2/Bax as compared with OGD/R group (P<0.05).Conclusion LBP treatment protects primary hippocampal neurons from OGD/R injury via inhibiting apoptosis and autophagic cell death.