1.2-APB inhibits H2O2-induced chondrocyte apoptosis through PKCα/HIF-1α 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 H2O2-induced chondro-cyte apoptosis and its mechanism.Methods The experiment was divided into control group,H2O2 group,2-APB group and H2O2+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 H2O2 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 H2O2-induced cells by 2-APB;Immunofluorescence was used to detect the fluorescent expression of HIF-1α in cells induced by H2O2 by PKCα inhibitor BIM-1.Results 2-APB inhibited H2O2-induced apoptosis in chon-drocytes,and the inhibitory effect was the most significant when the concentration of 2-APB was 100 pmol/L(F=235.80,P<0.01);22-APB could inhibit the positive rate of H2O2-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 H2O2.Conclusion 2-APB can inhibit chondrocytes apoptosis induced by H2O2 through the PKCα/HIF-1α pathway and thus protect chondro-cytes.