Mechanism Exploration on Emodin Ameliorates Cisplatin-induced Renal Tubular Cell Injury through Activation of Autophagy / 世界科学技术-中医药现代化

ABSTRACT

This study was aimed to observe the effect of emodin on cisplatin-induced renal tubular epithelial cells (NRK-52E) injury,in order to explore its possible molecular mechanisms.Firstly,effects of emodin on cisplatin-induced morphological changes in NRK-52E cells were observed.Secondly,the apoptosis-related protein expression of Caspase-3 and cleaved Caspase-3 were detected after the treatment of cisplatin alone or cisplatin together with emodin by western blot.Then,the expression of microtubule-associated protein 1 light chain 3 (LC3) Ⅱ/Ⅰ was detected after the treatment of emodin or rapamycin at different time points by western blot.Changes of pmRFP-LC3 fluorescent particles were observed by fluorescence microscopy.And effects of rapamycin on cellular morphological changes were observed in the environment of cisplatin.Finally,effects of emodin on the activation of AMPK and mTOR signal pathway were further observed,which is considered as the upstream of autophagy signaling pathway.The results showed that cisplatin can induce morphological changes in NRK-52E cell,which was obviously ameliorated by the intervention of emodin.Additionally,the increased protein expression of cleaved Caspase-3 induced by cisplatin was obviously reduced after the intervention of emodin.The LC3-Ⅱ/LC3-Ⅰ ratio was significantly increased after the treatment of emodin or rapamycin at different time points.Rapamycin can significantly ameliorate NRK-52E cell apoptosis induced by cisplatin.Simultaneously,the number of pmRFP-LC3 fluorescent particles increased after the treatment of emodin.As the extension of time by intervention of emodin,the protein expression of p-mTOR was significantly reduced.In contrast,the protein expression of p-AMPK was significantly increased.It was concluded that emodin can ameliorate cisplatininduced apoptosis in NRK-52E cells.Its potential mechanism may be attributed to the activation of autophagy by regulating AMPK/mTOR signaling pathway.And thus,it played a role in renal protective effects.