MENG Weiyang,WU Fangfang,JIN Can, et al. The mechanism of Cu-ATSM in protecting human umbilical vein endothelial cells from oxygen-glucose deprivation and reperfusion injury[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2023, 53(2): 102-108.
Abstract：Objective: To investigate the effect of copper-diacetyl-2 (N4-methylthiosemicarourea) (Cu-ATSM) on human umbilical vein endothelial cells (HUVECs) apoptosis and its underlying mechanisms after oxygen-glucose deprivation/reperfusion (OGD/R). Methods: HUVECs were divided as the control, OGD/R and OGD/R+Cu-ATSM groups. Flow cytometry was used to detect the effect of Cu-ATSM on OGD/R induced cell apoptosis. Mito-tracker staining was used to detect the effect of Cu-ATSM on mitochondrial morphology. JC-1 staining was used to detect the change of mitochondrial membrane potential. Western blot was used to detect the expression of autophagy related proteins such as ATG5, Beclin1 and LC3I/II. Immunofluorescenc was used to detect the expression of LC3II. Results: Flow cytometry results showed that the number of HUVECs apoptosis was significantly increased in OGD/R group, compared with the control group. However, after Cu-ATSM treatment the apoptosis cells were significantly decreased (P<0.05), indicating that Cu-ATSM treatment inhibited the HUVECs cell apoptosis after OGD/R. Mito-tracker and JC-1 staining results suggested that mitochondrial morphology and function were significantly damaged and membrane potential decreased in the OGD/R group,compared with the control group. However, the Cu-ATSM treatment ameliorated mitochondrial function and maintained mitochondrial morphology and mitochondrial membrane potential. Western blot results suggested that compared with the control group, the expression of autophagy related protein such as ATG5, Beclin1 and LC3II increased in OGD/R group, while the expression of autophagy related proteins decreased after Cu-ATSM treatment (P<0.05). Immunofluorescence results showed that compared with the control group, the number of LC3II was significantly increased after OGD/R injury, while the number of LC3II was significantly decreased after Cu-ATSM treatment. Conclusion: Cu-ATSM improves mitochondrial function and inhibits apotosis on HUVECs after OGD/R, which may be achieved by inhibiting autophagy.
