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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研究
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| 篇名： | 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研究 |
| TITLE： | |
| 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300 μmol/L过氧化氢（H2O2）在37 ℃孵育8 h；二氮嗪组细胞先以300 μmol/L二氮嗪在37 ℃预处理0.5 h后，再以300 μmol/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。 |
| 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 3rd 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 （H2O2） 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 H2O2 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. |
| 期刊： | 2019年第30卷第14期 |
| 作者： | 蒋翠云，任少琳，张纯萍，程少文 |
| AUTHORS： | JIANG Cuiyun，REN Shaolin，ZHANG Chunping，CHENG Shaowen |
| 关键字： | 二氮嗪；氧化损伤；软骨细胞；增殖活力；凋亡；内质网应激途径；机制 |
| KEYWORDS： | Diazoxide； Oxidative injury； Chondrocytes； Proliferation activity； Apoptosis； Endoplasmic reticulum stress pathway； Mechanism |
| 阅读数： | 271 次 |
| 本月下载数： | 5 次 |
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