1.2-APB inhibits H2 O2 -induced chondrocyte apoptosis through PKCα/ HIF-1 α signaling pathway
Ziwei Ouyang ; Lei Dong ; Yan Wang ; Yuanzhi Cheng ; Rendi Zhu ; Renpeng Zhou ; Yingjie Zhao ; Wei Hu
Acta Universitatis Medicinalis Anhui 2024;59(7):1150-1156
Objective :
To explore the effect of 2-aminoethoxy-diphenyl borate(2-APB) on H2 O2 -induced chondro- cyte apoptosis and its mechanism.
Methods :
The experiment was divided into control group,H2 O2 group,2-APB group and H2 O2 + 2-APB group.CCK-8 method was used to detect the cell viability of each group ; The effect of 2- APB on the morphological changes of chondrocytes induced by H2 O2 was observed under microscopy ; TUNEL meth- od and flow cytometry were used to detect chondrocyte apoptosis ; Flow cytometry was used to detect Lipid reactive oxygen species ( ROS) ; Western blot was used to detect the protein expressions of Cleaved-PARP,p-PKCα and HIF-1α in H2 O2 -induced cells by 2-APB ; Immunofluorescence was used to detect the fluorescent expression of HIF- 1 α in cells induced by H2 O2 by PKCα inhibitor BIM-1 .
Results :
2-APB inhibited H2 O2 -induced apoptosis in chon- drocytes,and the inhibitory effect was the most significant when the concentration of 2-APB was 100 μmol / L (F = 235. 80,P < 0. 01 ) ; 22-APB could inhibit the positive rate of H2 O2 -induced apoptosis of chondrocytes ( F = 114. 80,P<0. 01) and the level of ROS (F = 52. 99,P<0. 01) .and inhibited the expression of Cleaved-PARP (F = 10. 10,P<0. 05) ,p-PKCα (F = 24. 56,P<0. 05) and HIF-1α proteins (F = 6. 85,P<0. 05) .The PKCα in- hibitor BIM-Ⅰ could inhibit the increase in HIF-1α fluorescence intensity caused by H2 O2.
Conclusion
2-APB can inhibit chondrocytes apoptosis induced by H2 O2 through the PKCα/ HIF-1α pathway and thus protect chondro- cytes.