依达拉奉对抗化学性缺氧诱导的HEI-OC1听细胞内质网应激性凋亡  

Protection against CoCl2-induced endoplasmic reticulum stress and apoptosis in HEI-OC1 cells by edaravone

作　　者：曹磊[1] 张世平[1] 胡亿文[1] 

机构地区：[1]广州市红十字会医院耳鼻喉科,广州510220

出　　处：《中华耳科学杂志》2011年第1期87-91,共5页Chinese Journal of Otology

摘　　要：目的探讨依达拉奉(edaravone,EDA)能否保护HEI-OC1(House Ear Institute-organ of Corti 1)听细胞对抗氯化钴(CoCl2)诱导的内质网应激性凋亡。方法用CoCl2处理HEI-OC1听细胞,建立化学性缺氧损伤HEI-OC1听细胞的体外模型。EDA在CoCl2处理细胞前1h加入培养基中作为预处理。细胞计数试剂盒-8(cell count kit 8,CCK-8)比色法检测细胞存活率;试剂盒法检测细胞内丙二醛(Malondialdehyde,MDA)的含量;Western blot法(蛋白印迹法)检测葡萄糖调节蛋白78(glucose regulating protein 78,GRP78;或称内质网应激蛋白)及裂解型半胱氨酸天冬氨酸蛋白酶-12(cleaved caspase-12,活化形式)的表达。结果 CoCl2在100～400μmol/L浓度范围内处理HEI-OC1听细胞24h,可浓度依赖性地降低细胞活力;300μmol/LCoCl2处理HEI-OC1听细胞,可以上调内质网应激蛋白GRP78的表达、增加细胞内MDA的含量及活化caspase-12。在300μmol/LCoCl2作用HEI-OC1听细胞前,用10～40μmol/LEDA预处理1h可以浓度依赖性地抑制CoCl2诱导的毒性损伤作用,40μmol/LEDA预处理1h可抑制CoCl2对GRP78表达的上调和对caspase-12的活化作用,以及CoCl2引起的MDA含量增加。结论 EDA可通过抗氧化及抗内质网应激引起的凋亡以保护HEI-OC1听细胞对抗CoCl2诱导的细胞损伤。Objective To study whether edaravone （EDA） can protect HEI-OC1 （House Ear Institute-organ of Corti 1） auditory cells （HEI-OC1 cells） against CoCl2-induced endoplasmic reticulum stress （ERS） and apoptosis. Methods HEI-OC1 cells were treated with CoCl2 as a model of hypoxic auditory cellular toxicity. EDA was added into the culture medium before CoCl2 treatment as a pretreatment. Cell viability was measured with cell counter kit （CCK-8）. Intracellular malondialdehyde （MDA） level was detected by a commercial kit. The expressions of glucose regulating protein 78 （GRP78） and cleaved caspase-12 were evaluated by Western blot assay. Results Exposure of HEI-OC1 cells to CoCl2 at 100-400 μmol/L for 24 h decreased cell viability in a dose-dependent manner. Pre-treatment with 300 μmol/L CoCl2 resulted in over-expressions of GRP78 and cleaved caspase-12, as well as an increase in intracellular MDA level in HEI-OC1 cells. Pretreatment of EDA at 10-40 μmol/L inhibited CoCl2 treatment-induced cytotoxicity in a concentration-dependently manner. Pretreatment of 40 μmol/L EDA suppressed 300 μmol/L CoCl2 treatment caused-upregulations of GRP78 and cleaved caspase-12, as well as the increase in intracellular MDA level. Conclusion EDA can protect HEI-OC1 cells against CoCl2-induced injury, via anti-oxidation and anti-ERS -mediated apoptosis

关 键 词：依达拉奉 化学性缺氧 内质网应激 氧化应激 凋亡 HEI-OC1听细胞 

分 类 号：R764.35[医药卫生—耳鼻咽喉科] R364.4[医药卫生—临床医学]