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

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2-APB inhibits H2 O2 -induced chondrocyte apoptosis through PKCα/ HIF-1 α signaling pathway

Author: Ziwei Ouyang ^{1
,

2} ; Lei Dong ³ ; Yan Wang ^{1
,

2} ; Yuanzhi Cheng ^{1
,

2} ; Rendi Zhu ^{1
,

2} ; Renpeng Zhou ^{1
,

2} ; Yingjie Zhao ^{1
,

2} ; Wei Hu ^{1
,

2}
Author Information

1. School of Pharmacy，Anhui Medical University，Hefei 230032
2. Dept of Clinical Pharmacology， The Second Hospital of Anhui Medical University，Hefei 230601
3. Dept of Clinical Pharmacology， The Second Hospital of Anhui Medical University，Hefei 230601
Publication Type:Journal Article
Keywords: 2-aminoethoxy-diphenyl borate; chondrocytes; apoptosis; H2 O2; PKCα/ HIF-1α signaling pathway
From: Acta Universitatis Medicinalis Anhui 2024;59(7):1150-1156
CountryChina
Language:Chinese
Abstract: 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.
Full text:20240910163859521162-APB通过PKCα_H...2O_2诱导的软骨细胞凋亡_欧阳紫微.pdf