Long Zonghong, Duan Guangyou, Li Hong, Yi Tingting, Wu Xiaoxiao, Chen Feng, Wu Zhuoxi, Gao Yuqi
Department of Anesthesiology, Xinqiao Hospital, Third Military Medical University, Chongqing, China.
Key Laboratory of High Altitude Medicine, Third Military Medical University, Chongqing, China.
PLoS One. 2017 Jul 27;12(7):e0181903. doi: 10.1371/journal.pone.0181903. eCollection 2017.
This study aimed to use long-term diazoxide treatment to establish a loss-of-cardioprotection model and then perform proteomics analysis to explore which proteins of mitochondrial inner membrane (MIM) are potentially involved in delayed cardioprotection. Rats received 1 to 8 weeks of diazoxide treatments (20 mg•kg-1•d-1) to establish a loss-of-cardioprotection model in different groups. Detection of serum cTnI levels and cell apoptosis assays in heart tissue were performed. Then, rats MIM after 0, 4 and 6 weeks of diazoxide treatment was isolated and proteomics analysis was performed. An invitro model of H9C2 cells was performed to explore the effects of targeted protein on delayed cardioprotection. The effect of delayed cardioprotection by diazoxide preconditioning disappeared when diazoxide treatments were given for six weeks or longer. Ubiquinol-cytochrome c reductase core protein 1 (UQCRC1) was identified in the proteomics analysis. UQCRC1 expression was upregulated by diazoxide treatment in H9C2 cells, and UQCRC1 down-regulation could increase the lactate dehydrogenase release and apoptosis rate after injury induced by oxygen glucose deprivation. These results showed that UQCRC1 might contribute to the loss-of-cardioprotection model induced by long-term diazoxide treatment and play a role in delayed cardioprotection.
本研究旨在通过长期使用二氮嗪治疗建立心脏保护作用丧失模型，然后进行蛋白质组学分析，以探索线粒体内膜（MIM）的哪些蛋白质可能参与延迟性心脏保护。大鼠接受1至8周的二氮嗪治疗（20mg•kg-1•d-1），以在不同组中建立心脏保护作用丧失模型。进行血清肌钙蛋白I水平检测和心脏组织细胞凋亡分析。然后，分离二氮嗪治疗0、4和6周后的大鼠MIM并进行蛋白质组学分析。建立H9C2细胞的体外模型，以探索靶向蛋白对延迟性心脏保护的影响。当给予二氮嗪治疗六周或更长时间时，二氮嗪预处理的延迟性心脏保护作用消失。在蛋白质组学分析中鉴定出泛醇 - 细胞色素c还原酶核心蛋白1（UQCRC1）。在H9C2细胞中，二氮嗪处理上调UQCRC1表达，UQCRC1下调可增加氧糖剥夺诱导损伤后的乳酸脱氢酶释放和凋亡率。这些结果表明，UQCRC1可能导致长期二氮嗪治疗诱导的心脏保护作用丧失模型，并在延迟性心脏保护中发挥作用。
