本研究旨在探讨PPARγ/CPT-1调控对顺铂诱导的耳蜗毛细胞损伤的保护作用。生存能力,CCK-8法测定顺铂诱导的HEI-OC1细胞凋亡和线粒体膜电位,TUNEL和JC-1染色,分别。用MDA检测试剂盒检测氧化应激和脂质代谢,ROS,SOD,CAT,TG和FFA。通过RT-qPCR检测过表达(OV)-PPARG和OV-CPT1A的转染效率,并通过Westernblot检测凋亡和脂质代谢相关蛋白的表达。因此,不同浓度(5、10、30μM)的顺铂抑制了活力,促进HEI-OC1细胞凋亡并阻碍线粒体功能,伴随着Bax和caspase-3的上调表达和Bcl-2的下调表达。顺铂(5,10,30μM)诱导氧化应激加重,脂质代谢受到抑制,MDA水平的增加证明了这一点,ROS,TG,FFA和SOD和CAT水平降低。PPARG或CPT1A的过表达可以提高小鼠的生存力,线粒体功能,脂质代谢并抑制顺铂诱导的HEI-OC1细胞的氧化应激和凋亡。总之,PPARG或CPT1A的上调通过改善细胞脂质代谢和抑制氧化应激改善耳蜗毛细胞损伤。
This study aimed to investigate the protective effects of PPARγ/CPT-1 regulation on cisplatin-induced cochlear hair cell injury. The viability, apoptosis and mitochondrial membrane potential of cisplatin-induced HEI-OC1 cells were determined by CCK-8 assay, TUNEL and JC-1 staining, respectively. The oxidative stress and lipid metabolism were detected by the assay kits of MDA, ROS, SOD, CAT, TG and FFA. The transfection efficiency of overexpression (OV)-PPARG and OV-CPT1A was examined by RT-qPCR and the expressions of apoptosis- and lipid metabolism-related proteins were detected by western blot. As a result, cisplatin with varying concentrations (5, 10, 30 μM) suppressed the viability, promoted the apoptosis and hindered the mitochondrial function of HEI-OC1 cells, accompanied with up-regulated expressions of Bax and cleaved caspase-3 and down-regulated expression of Bcl-2. The oxidative stress was aggravated and lipid metabolism was inhibited by cisplatin (5, 10, 30 μM) induction, evidenced by the increased levels of MDA, ROS, TG, FFA and the decreased levels of SOD and CAT. Overexpression of PPARG or CPT1A could improve the viability, mitochondrial function, lipid metabolism and suppress the oxidative stress and apoptosis of cisplatin-induced HEI-OC1 cells. In conclusion, up-regulation of PPARG or CPT1A ameliorated cochlear hair cell injury by improving cellular lipid metabolism and inhibiting oxidative stress.