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目的观察氯化钴(CoCl2)对血清-葡萄糖剥夺(SGD)诱导的H9c2心肌细胞损伤的影响,探讨热休克蛋白90(HSP90)在其中的作用。方法用SGD的方法处理H9c2心肌细胞,建立缺血性损伤的心肌细胞模型。在SGD过程中同时给予CoCl2处理;在应用SGD和CoCl2之前给予HSP90抑制剂(17-AAG)预处理。处理结束后,检测细胞存活率、HSP90的表达、细胞内活性氧(ROS)以及线粒体膜电位(ΔΨm)。结果 SGD处理可引起H9c2心肌细胞损伤,表现为细胞存活率降低、胞内ROS水平升高及ΔΨm丢失。SGD处理还可降低H9c2心肌细胞内HSP90的表达。在50～100μmol.L-1浓度范围内,CoCl2处理可保护H9c2心肌细胞对抗SGD引起的存活率降低,100μmol.L-1CoCl2还可对抗SGD引起的ROS水平升高和ΔΨm丢失。100μmol.L-1CoCl2可时间依赖性地促进胞内HSP90的表达。在50～200μmol.L-1浓度范围内,CoCl2处理可对抗SGD诱导的HSP90表达下调,其中100μmol.L-1的CoCl2具有最强的拮抗作用。17-AAG通过抑制HSP90的作用可明显减弱CoCl2诱导的上述心肌细胞保护作用。结论 CoCl2可保护H9c2心肌细胞对抗SGD引起的损伤,其机制之一与上调HSP90表达有关。
Objective To observe the effect of cobalt chloride (CoCl2) on the injury of H9c2 cardiomyocytes induced by serum-glucose deprivation (SGD) and to explore the role of HSP90 in it. Methods H9c2 cardiomyocytes were treated with SGD to establish a cardiomyocyte model of ischemic injury. CoCl2 treatment was administered concurrently with SGD; HSP90 inhibitor (17-AAG) pretreatment was administered prior to application of SGD and CoCl2. After treatment, cell viability, HSP90 expression, intracellular reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) were measured. Results SGD treatment can cause H9c2 cardiomyocyte injury, which is characterized by decreased cell viability, increased intracellular ROS and loss of ΔΨm. SGD treatment also decreased HSP90 expression in H9c2 cardiomyocytes. In the concentration range of 50 ~ 100μmol.L-1, CoCl2 treatment can protect H9c2 cardiomyocytes against SGD-induced decreased survival, 100μmol.L-1CoCl2 against SGD-induced ROS levels rise and ΔΨm loss. 100μmol.L-1CoCl2 promoted the expression of intracellular HSP90 in a time-dependent manner. In the concentration range of 50 ~ 200μmol.L-1, CoCl2 treatment could counteract the down-regulation of HSP90 expression induced by SGD, of which 100μmol.L-1 CoCl2 had the strongest antagonistic effect. 17-AAG significantly attenuated CoCl2-induced cardioprotection by inhibiting HSP90. Conclusion CoCl2 protects H9c2 cardiomyocytes against SGD-induced injury, one of the mechanisms is related to the up-regulation of HSP90 expression.