| OBJECTIVE:To investigate the cardioprotective effects and mechanism of Cyclovirobuxine D on neonatal rat cardiomyoeytes exposed to hypoxia-reoxygenation (HR) injuryMETHODS:The hypoxia-reoxygenation injury model of cultured neonatal rat cardiomyocytes was built.Primary cultured cardiomyocytes were randomly divided into four groups:control group;H/R(hypoxia-reoxygenation) group:cardiomyocytes were incubated with 3%02,5%CO2,92%N2 for 16h,then with 37℃,5%CO2 for 4h;CVB-D group(1×10-5mol/L):cardiomyocytes were incubate with CVB-D (1×10-5mol/L) for 4 h;CVB-D+H/R group:CVB-D was administered 30 min before treatment with 3%02,5%CO2,92%N2 for 16 h,then with 37℃,5%CO2 for 4 h.The activity of lactate dehydrogenase(LDH) in the culture medium was detected,the contents of MDA,an indicator for lipid peroxidation,and the activity levels of the antioxidase SOD of the culture medium were measured,the apoptosis rate of cardiomyocyte were examined with FCM.The activity of Caspase-3 was evaluated by Caspase-Glo(?)3/7 Reagent.RESULTS:1.Activity of LDH:Compared with control group,the activity of LDH was increased significantly in H/R group(227.2±14.2 vs 97.8±13.0 U/L,P＜0.01). Compared with H/R group,the activity of LDH was decreased in CVB-D+H/R group significantly(126.0±23.8 vs 227.2±14.2 U/L,P＜0.01).There was no difference in the activity of LDH between CVB-D group and control group.2.Apoptosis rate:Compared with control group,the apoptosis rate of cardiomyocytes was increased significantly in H/R group(13.2±1.1 vs 0.7±0.1%P＜0.01);Compared with H/R group,the apoptosis rate of cardiomyocytes was decreased in CVB-D+H/R group significantly(3.0±0.3 vs 13.2±1.1%P＜0.01).There was no difference in the apoptosis rate of cardiomyocytes between CVB-D group and control group.3.Content of MDA and activity SOD;Compared with control group,the content of MDA increased significantly in H/R group(3.21±0.17 vs 0.76±0.13μmol·L-1 P＜0.01),while the activity of SOD obviously decreased in H/R group (284.30±13.20 vs 165.22±11.67 U·ml-1 P＜0.01).Compared with H/R group,the CVB-D+H/R group treated with CVB-D(1×10-5mol/L) markedly attenuated the increase in MDA content(1.36±0.08 vs 3.21±0.17μmol·L-1 P＜0.01),whereas the levels of SOD activity were obviously increased in the CVB-D+H/R group (267.40±14.52 vs 165.22±11.67 U·ml-1 P＜0.01).There was no difference in the activity of LDH between CVB-D group and control group.4.Activity of Caspase-3:The activity of Caspase-3 in H/R group was obviously higher than that of control group(1.42±0.07 vs 1.10±0.08 P＜0.01);Compared with H/R group,the activity of Caspase-3 was decreased in CVB-D+H/R group significantly(1.12±0.08 vs 1.42±0.07,P＜0.01).There was no difference in the activity of Caspase-3 between CVB-D group and control group.CONCLUSIONS:Primary cultured cardiomyocytes in neonatal rats were protected by Cyclovirobuxine D from injury of hypoxia-reoxygenation(H/R).The protective mechanism could be related to its ability to decrease the activity of LDH,reduce apoptosis rate and lipid peroxidation.
|
PDF Full Text Request