RNA-Seq Based Transcriptome Analysis of Ethanol Extract of Saffron Protective Effect Against Corticosterone-Induced PC12 Cell Injury

Abstract

Background Saffron is a traditional Chinese herbal medicine, which is typically used in clinical to regulate anxiety, tension, and other depression-related conditions. The study aimed to explore the neuroprotective effect of ethanol extract of saffron (EES) on corticosterone (CORT)-induced injury in PC12 cells and further explored its potential mechanism. Methods The authenticity of saffron and the active components of EES were identified by a water test and ultra-performance liquid chromatography-time of flight mass spectrometry system. The screening of cytotoxicity for PC12 cells was incubated with EES in different concentrations for 24 h, and the protective efficacy of EES on CORT (500 µM) induced PC12 cell injury, cell viability was assessed by Cell Counting Kit-8 (CCK-8) assay. The DEGs of EES-protected PC12 cells were analyzed using the RNA-seq method, and the results were analyzed for GO and KEGG enrichment. The results of RNA-seq were verified by qPCR analysis. Results The saffron was initially identified as authentic in the water test and 10 compounds were identified by UPLC-MS. The results of CCK-8 demonstrated that EES at concentrations above 640 µg/mL exerted a certain cytotoxic effect, and PC12 cells pretreated with EES (20, 40, and 80 µg/mL) significantly reversed the 500 µM CORT-induced cell death. RNA-seq analysis showed that EES regulated 246 differential genes, which were mainly enriched in the MAPK signaling pathway. Dusp5, Dusp6, Gadd45b, Gadd45G, and Pdgfc were further validated by qPCR. Experimental data showed that the results of qPCR were consistent with RNA-seq. Conclusions These findings provide an innovative understanding of the molecular mechanism of the protective effect of EES on PC12 cells at the molecular transcription level, and the above molecules may be potential novel targets for antidepressant treatment.