Protective Effects Of Dehydrocostuslactone On Oxygen-glucose Deprivation Injury In Rat Hippocampal Slices

Objective: To study the protective effects of dehydrocostuslactone on oxygen-glucose deprivation injury in rat hippocampal slices and to explore the mechanism of mitochondrial apoptosis.Materials and methods:We are subjected to the rat hippocampal slices induced by OGD model,the brain slices were randomly divided into blank group,model group,Nimodipine group(10 ?M),DHL low,middle and high dose group(1,5 and 10 ?M).The blank group was incubated with normal aCSF and continued to pass 95% O2 / 5% CO2.The model group,Nimodipine group,DHL low,medium and high dose group were treated with sugar-free aCSF for 30 min OGD,and then transferred to 95% O2 / 5% CO2 gas environment for 1 h.After 1 h,the supernatant was collected and the supernatant of LDH was incubated.The brain slices were incubated with 2% TTC at 37 ° C for 10 min,and 20 ?l/mg of the extract was added(Ethanol:DMSO = 1:1).Room temperature sealed dark for 24 h,measured OD490 absorbance value;collected each group of brain slices.The expression of Cyto-c,Apaf-1,Noxa,Mcl-1,Bcl-2,Bax and Caspase-9,Caspase-7 and Caspase-3 protein in brain slices were detected by Western-blot.The part of Caspases inhibitor was performed as described former,the brain slices were randomly divided into blank group,model group,Nimodipine group(10?M),Z-VAD-FMK group,Z-VAD-FMK + DHL high dose group.After the experiment,the expression of Cyto-c,Apaf-1,Caspase-9,Caspase-7 and Caspase-3 were detected by Western blot.Results: 1.Compared with the model group,the release of LDH was decreased by the three drug groups(p<0.05),and the absorbance of TTC was increased(p<0.05).The protein expression of Bax significantly decreased(p<0.05).The expression of Bcl-2 and Mcl-1 significantly increased(p<0.05).The protein expression of Caspase-9,Caspase-7,Caspase-3,Cyto-c,Apaf-1 and Noxa were significantly decreased(p<0.05).2.Compared with model group,the protein expression of Caspase-9,Caspase-7,Caspase-3,Cyto-c and Apaf-1 were significantly decreased in Z-VAD-FMK group and Z-VAD-FMK + DHL high dose group(p<0.05).Conclusions: 1.Dehydrocostuslactone can protect the hippocampus brain injury by oxygen glucose deprivation,the protective mechanism may be by reducing the release of LDH to improve mitochondrial respiratory enzyme activity,and down-regulation of Bax,Caspase-9,Caspase-7,Caspase-3,Cyto-c,Apaf-1 and Noxa and up-regulated Blc-2 and Mcl-1 protein expression.2.Dehydrocostuslactone may have the same mechanism with Caspase protease inhibitor Z-VAD-FMK.