Proteomic Analysis and the Antioxidant Activity Evaluation of Asini Corii Colla Powder Based on Liquid Chromatography-Tandem Mass Spectrometry Technology

Asini corii colla contains multiple bioactive proteins， endowing it with multifaceted health benefits. This study employed liquid chromatography-tandem mass spectrometry （LC-MS/MS） to systematically identify and analyze the protein composition of Asini corii colla powder， identifying various functional proteins including multiple intermediate filament proteins， collagens， immunomodulatory proteins， and antioxidant-related proteins. Further evaluation of Asini corii colla powder's cytotoxicity， proliferation， and migration effects was conducted using human dermal fibroblasts （HSF）. Results indicated no significant cytotoxicity within the tested concentration range （0.156~20 mg/mL）， while promoting HSF proliferation and migration， suggesting preliminary potential for Asini corii colla powder in skin repair applications. To investigate the antioxidant stress activity of Asini corii colla， this study further employed a hydrogen peroxide （H₂O₂） induced cellular oxidative stress model to evaluate the effects of Asini corii colla powder on intracellular reactive oxygen species （ROS） scavenging capacity and antioxidant enzyme activity. Experiments demonstrated that pre-treatment with Asini corii colla powder significantly suppressed H₂O₂-induced ROS levels while enhancing intracellular superoxide dismutase （SOD）， catalase （CAT）， and glutathione peroxidase （GSH-Px） activities. This confirms its antioxidant effects are associated with strengthening the endogenous antioxidant defense system. Proteomics analysis further revealed that Asini corii colla powder may regulate oxidative stress by modulating the expression of proteins associated with the heat shock protein B family， biliverdin reductase B， hemoglobin subunit α， and nuclear factor κB （NF-κB） signaling pathways， thereby mitigating cellular oxidative stress damage to a certain extent. In summary， this study elucidates the protein composition of Asini corii colla and validates its role and mechanisms in combating oxidative stress， laying a scientific foundation for expanding its applications in skin repair and anti-aging.