Antioxidative Effect Of Sanghuangporus Sanghuang Polysaccharides Based On AFM In Vitro

Oxidative damage can cause oxidative stress in the body,which can lead to a variety of diseases,such as aging,melanin deposition,skin cancer,and inflammation.Sanghuangporus sanghuang is a medicinal and food fungus,and it has been reported in the literature that Sanghuangporus sanghuang polysaccharides has the effect of scavenging DPPH free radicals.Therefore,this thesis proposes to investigate the in vitro antioxidant effect of Sanghuangporus sanghuang polysaccharides based on AFM(Atomic Force Microscopy)through a cellular model.AFM can detect the mechanical properties of cells at the nanoscale,such as cell height,roughness,adhesion,Young’s modulus,etc.AFM investigates the role of Sanghuangporus sanghuang polysaccharides in inhibiting oxidative damage in model cells by characterizing the mechanical properties of cellular physics.This study investigates the in vitro antioxidant effect of Sanghuangporus sanghuang polysaccharides based on the AFM technique,with the following main work:1)The extraction of Sanghuangporus sanghuang polysaccharides was carried out by aqueous extraction.Optimization of the extraction process of Sanghuangporus sanghuang polysaccharides by response surface methodology and screening of the optimal extraction method.The optimum extraction conditions: extraction time of 2 hours,material-liquid ratio of 1:30,and extraction temperature of 70℃.2)The preliminary antioxidant vitality of the extracted Sanghuangporus sanghuang polysaccharides was determined by DPPH free radical scavenging assay.A oxidative damage model was developed HaCaT cells as the study target.To study the antioxidant vitality of different concentrations of Sanghuangporus sanghuang polysaccharides treated on HaCaT cells for different times respectively.Screening of suitable experimental concentrations of Sanghuangporus sanghuang polysaccharides(low concentration: 7.5μg/m L,high concentration: 15 μg/m L)for further studies on the effects of Sanghuangporus sanghuang polysaccharides on the mechanical properties of HaCaT cells.3)The experiment based on the DPPH free radical scavenging assay and the preliminary assay of antioxidant vitality of Sanghuangporus sanghuang polysaccharides were based on the study of HaCaT cells.We analysis the the mechanical properties of HaCaT cells before and after oxidative damage by AFM.HaCaT cell morphology before and after oxidative damage were scanned by SEM.The skeleton of HaCaT cells before and after oxidative damage was scanned by fluorescent staining.In this way,the inhibitory effect of Sanghuangporus sanghuang polysaccharides on the antioxidant damage of HaCaT cells was investigated.The experimental results showed that the mechanical parameters(Young’s modulus,adhesion force,and surface roughness)of single cell was detected and analyzed,and the high and low concentrations of Sanghuangporus sanghuang polysaccharides has different degrees of effects on maintaining the mechanical properties of the cells,and had significantly inhibited the oxidative damage of ethanol on HaCaT cells.The mechanical parameters of ethanol oxidatively damaged cells,gradually approached those of control HaCaT cells with the increase of the action time of Sanghuangporus sanghuang polysaccharides.The results of the SEM and fluorescence staining experiments were consistent with the results of the AFM study,indicating that the mechanical parameters of the cells can be used as an indicator of cellular vitality.In this study,the antioxidant viability of Sanghuangporus sanghuang polysaccharides was investigated based on AFM,with a view to provide a research basis for the development and application of Sanghuangporus sanghuang,especially for applications in cosmetics and other fields.