Alterations of brain quantitative proteomics profiling revealed the molecular mechanisms of diosgenin against cerebral ischemia reperfusion effects.

Diosgenin (DIO), the starting material for the synthesis of steroidal anti-inflammatory drugs in pharmaceutical industry, has been previously demonstrated to display pharmaceutical effects against cerebral ischemic reperfusion (I/R). However, the alterations of brain proteome profiles underlying this treatment remain elusive. In the present study, the proteomics analysis of the brain tissues from I/R rats after DIO treatment was performed by an integrated TMT-based quantitative proteomic approach coupled with LC-MS/MS technology. A total of 5043 proteins (ProteomeXchange identifier: PXD016303) were identified, of which 58 common differentially expressed proteins (DEPs) were significantly dysregulated in comparison between Sham verse I/R and I/R verse DIO. The 8 validated proteins including EPG5, STAT2, CPT1A, EIF2AK2, GGCT, HIKESHI, TNFAIP8, and EMC6 by qPCR and Western blotting consistently supported the TMT-based proteomic results, which were mainly associated with autophagy and inflammation response. Considering the anti-inflammatory characters of DIO, the biological functions of STAT2 and HIKESHI that are the probably direct anti-inflammatory targets were further investigated during the course of I/R treated with DIO. In addition, the combination of verified STAT2 and HIKESHI in peripheral blood samples from stroke patients resulted in AUC value of 0.765 with P<0.004 to distinguish stroke patients from healthy controls. Taken together, the current findings firstly mapped comprehensive proteomic changes after I/R treated with DIO to better decipher the molecular mechanisms mainly based on anti-inflammatory aspect underlying this therapeutic effect, providing a foundation for developing potentially therapeutic targets of anti-I/R of DIO and clinically prognostic biomarkers of stroke.