【摘要】 目的:基于内质网应激(ERS)途径探讨二氮嗪对氧化损伤状态下软骨细胞增殖和凋亡的影响机制。方法:取SD小鼠膝关节软骨细胞进行原代培养,取第3代软骨细胞随机分为对照组、损伤模型组和二氮嗪组。对照组细胞不作处理;损伤模型组细胞以300μmol/L过氧化氢(H2O2)在37℃孵育8 h;二氮嗪组细胞先以300μmol/L二氮嗪在37℃预处理0.5 h后,再以300μmol/L H2O2同法孵育8 h。采用CCK-8法检测细胞的增殖活力,采用流式细胞术检测细胞的凋亡率,采用Western blotting法检测细胞中ERS相关蛋白[胱天蛋白酶3(Caspase-3)、Bcl-2相关X蛋白(Bax)、增强子结合蛋白环磷酸腺苷反应元件结合转录因子同源蛋白(CHOP)]的表达情况。结果:与对照组比较,损伤模型组细胞增殖活力显著降低,凋亡率显著升高(P<0.05);Caspase-3、Bax、CHOP蛋白表达量均显著升高(P<0.05)。与损伤模型组比较,二氮嗪组软骨细胞增殖活力显著升高,凋亡率显著降低(P<0.05);上述相关蛋白表达量均显著降低(P<0.05)。结论:二氮嗪可能通过抑制ERS途径,提高软骨细胞在氧化损伤状态下的增殖活力,并抑制细胞凋亡。更多还原

【Abstract】 OBJECTIVE:To investigate the effect mechanism of diazoxide on the proliferation and apoptosis of chondrocytes with oxidative injury based on endoplasmic reticulum stress（ERS） pathway. METHODS:SD mice were selected for primary culture of articular chondrocytes. The 3 rd generation chondrocytes were randomly divided into control group,injury model group and diazoxide group. Control group didn’t receive any treatment. The injury model group was incubated with 300 μmol/L hydrogen peroxide（H₂ O₂）at 37 ℃ for 8 h. Diazoxide group was pretreated with 300 μmol/L diazoxide at 37 ℃ for 0.5 h,and then incubated with 300 μmol/L H₂ O₂ for 8 h. The proliferation of chondrocytes was detected by CCK-8 assay. The apoptosis rate of chondrocytes was detected by flow cytometry. The expression of ERS-related proteins [Caspase-3,Bcl-2-associated X（Bax）,C/EBP homologous protein（CHOP）] were detected by Western blotting assay. RESULTS:Compared with control group,the proliferation activity of chondrocytes in injury model group was significantly decreased,while apoptosis rate was increased significantly（P<0.05）;the protein expression of Caspase-3,Bax and CHOP were increased significantly（P<0.05）. Compared with injury model group,the proliferation activity of chondrocytes in diazoxide group was increased significantly,while the apoptosis rate was decreased significantly（P<0.05）; the expression of above related proteins were decreased significantly（P<0.05）. CONCLUSIONS:Diazoxide can improve the proliferation activity of chondrocytes with oxidative injury and inhibit their apoptosis by inhibiting ERS pathway.更多还原