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 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 P < 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.