iProXProteomicsZengjun GuoRattus Rattushttp://www.iprox.org/page/project.html?id=IPX0001872000guozj@mail.xjtu.edu.cnCollege of Pharmacy, Xi’an Jiaotong UniversityiProXDiosgenin (DIO), the starting material for the synthesis of steroidal anti-inflammatory drugs in the 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 using an integrated TMT-based quantitative proteomic approach coupled with the liquid chromatography with tandem mass spectrometry technology. A total of 5043 proteins (ProteomeXchange identifier: PXD016303) were identified, of which 58 common differentially expressed proteins were significantly dysregulated in comparison between sham versus I/R and I/R versus DIO. The eight validated proteins including EPG5, STAT2, CPT1A, EIF2AK2, GGCT, HIKESHI, TNFAIP8, and EMC6 by quantitative polymerase chain reaction 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 probable 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 the area under the curve value of 0.765 with <i>P</i> < 0.004 to distinguish stroke patients from healthy controls. Taken together, the current findings first mapped comprehensive proteomic changes after I/R was treated with DIO to better decipher the molecular mechanisms mainly based on the 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.Alterations of Brain Quantitative Proteomics Profiling Revealed the Molecular Mechanisms of Diosgenin against Cerebral Ischemia Reperfusion Effects.Zhang Xinxin X, Wang Xingbin X, Khurm Muhammad M, Zhan Guanqun G, Zhang Hui H, Ito Yoichiro Y, Guo Zengjun ZfalseAlterations of brain quantitative proteomics profiling revealed the molecular mechanisms of diosgenin against cerebral ischemia reperfusion effectsDiosgenin (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 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 strokeMon Nov 18 00:00:00 GMT 2019PXD0163031011731940440