2-APB通过PKCα/HIF-1α信号通路抑制H_(2)O_(2)诱导的软骨细胞凋亡  

2-APB inhibits H_(2)O_(2)-induced chondrocyte apoptosis through PKCα/HIF-1αsignaling pathway

作　　者：欧阳紫微 董雷 王琰 程远志 朱仁弟 周仁鹏[1,2] 赵英杰 胡伟 Ouyang Ziwei;Dong Lei;Wang Yan;Cheng Yuanzhi;Zhu Rendi;Zhou Renpeng;Zhao Yingjie;Hu Wei(School of Pharmacy,Anhui Medical University,Hefei 230032;Dept of Clinical Pharmacology,The Second Hospital of Anhui Medical University,Hefei 230601)

机构地区：[1]安徽医科大学药学院,合肥230032 [2]安徽医科大学第二附属医院药物临床试验研究中心,合肥230601

出　　处：《安徽医科大学学报》2024年第7期1150-1156,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82071591、82371575、82204405);安徽省自然科学基金(编号:2108085QH383)。

摘　　要：目的探讨2-氨基乙氧基苯硼酸(2-APB)对H_(2)O_(2)诱导的软骨细胞凋亡的作用及其机制。方法实验分为Control组、H_(2)O_(2)组、2-APB组和H_(2)O_(2)+2-APB组。CCK-8法检测各组细胞活力;显微镜下观察2-APB对H_(2)O_(2)诱导的软骨细胞形态变化的影响;TUNEL法和流式细胞术检测软骨细胞凋亡情况;流式细胞术检测脂质活性氧(ROS)水平;Western blot法检测2-APB对H_(2)O_(2)诱导的各组细胞中Cleaved-PARP、p-PKCα和HIF-1α蛋白的表达情况;免疫荧光法检测PKCα抑制剂BIM-Ⅰ对H_(2)O_(2)诱导的各组细胞中HIF-1α的荧光表达情况。结果2-APB对H_(2)O_(2)诱导的软骨细胞凋亡具有抑制作用,且当2-APB浓度为100μmol/L时抑制效果最为显著(F=235.80,P<0.01);2-APB能够抑制H_(2)O_(2)所致软骨细胞凋亡阳性率(F=114.80,P<0.01)以及ROS的水平(F=52.99,P<0.01),并且抑制Cleaved-PARP(F=10.10,P<0.05)、p-PKCα(F=24.56,P<0.05)和HIF-1α(F=6.85,P<0.05)蛋白的表达;PKCα抑制剂BIM-Ⅰ能够抑制H_(2)O_(2)所致HIF-1α荧光强度的增加。结论2-APB可以通过抑制PKCα/HIF-1α通路减少H_(2)O_(2)诱导的软骨细胞凋亡,进而保护软骨细胞。Objective To explore the effect of 2-aminoethoxy-diphenyl borate(2-APB)on H_(2)O_(2)-induced chondrocyte apoptosis and its mechanism.Methods The experiment was divided into control group,H_(2)O_(2)group,2-APB group and H_(2)O_(2)+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 H_(2)O_(2)was observed under microscopy;TUNEL method 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-PKCαand HIF-1αin H_(2)O_(2)-induced cells by 2-APB;Immunofluorescence was used to detect the fluorescent expression of HIF-1αin cells induced by H_(2)O_(2)by PKCαinhibitor BIM-1.Results 2-APB inhibited H_(2)O_(2)-induced apoptosis in chondrocytes,and the inhibitory effect was the most significant when the concentration of 2-APB was 100μmol/L(F=235.80,P<0.01);22-APB could inhibit the positive rate of H_(2)O_(2)-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-1αproteins(F=6.85,P<0.05).The PKCαinhibitor BIM-Ⅰcould inhibit the increase in HIF-1αfluorescence intensity caused by H_(2)O_(2).Conclusion 2-APB can inhibit chondrocytes apoptosis induced by H_(2)O_(2)through the PKCα/HIF-1αpathway and thus protect chondrocytes.

关 键 词：2-氨基乙氧基苯硼酸 软骨细胞 细胞凋亡 H_(2)O_(2) PKCα/HIF-1α通路 

分 类 号：R966[医药卫生—药理学]