ABSTRACT
Objective To investigate the protective effects and possible mechanism of pigment epithelium derived faetor (PEDF) on myocardial cells H9C2 under hypoxia and serum-free condition.Methods H9C2 cells were culture in vitro and performed the hypoxia and serum-free processing.The cells were divided into the control group (H9C2),hypoxia group (hypoxia + H9C2),PEDF group(hypoxia+H9C2 +PEDF) and mitochondrial fission inhibitor(Mdivi-1) group(hypoxia+h9C2+Mivi-1).The apoptotic rate was detected by TUNNEL staining.The proteins levels of dynamin related peptide1 (Drp1) and cleaved-caspase 3 were measured by Western blot.Electron Microscopy and MitoTracker Red were used to detect the mitochondria morphology,the mitochondrial membrane potential was evaluated by cationic dye JC-1.MitoSOXTM was used to detect mitochondrial reactive oxygen species (ROS).Results Hypoxia induced mitochondrial fission(P＜0.05).The hypoxia group (6 h) and control group had statistical difference(P＜0.05).PEDF reduces mitochondrial fission under hypoxia condition(P＜0.05),which had statistical difference between the PEDF group and hypoxia group (6 h)(P＜0.05).PEDF and Mdivi-1 could decrease cell apoptosis under hypoxia condition(24 h),compared with the hypoxia group,the difference was statistically significant(P＜0.05).Conclusion PEDF decrease cell apoptosis by inhibiting H9C2 cells mitochondrial fission under hypoxia condition.
ABSTRACT
Objective To investigate the protective effects and possible mechanism of pigment epithelium derived faetor (PEDF) on myocardial cells H9C2 under hypoxia and serum-free condition.Methods H9C2 cells were culture in vitro and performed the hypoxia and serum-free processing.The cells were divided into the control group (H9C2),hypoxia group (hypoxia + H9C2),PEDF group(hypoxia+H9C2 +PEDF) and mitochondrial fission inhibitor(Mdivi-1) group(hypoxia+h9C2+Mivi-1).The apoptotic rate was detected by TUNNEL staining.The proteins levels of dynamin related peptide1 (Drp1) and cleaved-caspase 3 were measured by Western blot.Electron Microscopy and MitoTracker Red were used to detect the mitochondria morphology,the mitochondrial membrane potential was evaluated by cationic dye JC-1.MitoSOXTM was used to detect mitochondrial reactive oxygen species (ROS).Results Hypoxia induced mitochondrial fission(P＜0.05).The hypoxia group (6 h) and control group had statistical difference(P＜0.05).PEDF reduces mitochondrial fission under hypoxia condition(P＜0.05),which had statistical difference between the PEDF group and hypoxia group (6 h)(P＜0.05).PEDF and Mdivi-1 could decrease cell apoptosis under hypoxia condition(24 h),compared with the hypoxia group,the difference was statistically significant(P＜0.05).Conclusion PEDF decrease cell apoptosis by inhibiting H9C2 cells mitochondrial fission under hypoxia condition.