| Gingerol is the main active ingredient in ginger(Zingiber officinale Rose.).The previous research results showed that the content of gingerol decreased significantly during the processing of processed ginger,among which,the content of 6-gingerol decreased between 15% and 42%,while the content of shogaol increased significantly,and a new component of Zingerone was produced during the processing,which indicated that the "quality" and "quantity" of gingerol of ginger changed after the processing of dried ginger into processed ginger.However,the effect of processing on the transformation and activity of gingerol of processed ginger is not completely clear,which is worthy of further study.Objective:The conversion rule of 6-gingerol during the processing of processed ginger was clarified,and the reference was provided for the processing technology and processing mechanism of processed ginger.To investigate the protective effects of 6-shogaol,zingerone and 6-gingerol on HUVEC,and to provide the basis for the internal mechanism of pharmacodynamic changes after transformation.Method:1.Study on the transformation rule of 6-gingerol and the change of antioxidant activity during the processing of processed gingerThe contents of 6-shogaol,zingerone and 6-gingerol in dried ginger and processed ginger samples of different sanding processes were determined by UPLC.The contents of 6-gingerol and its transformed products were investigated under different time and temperature conditions.The in vitro antioxidant activities of processed ginger samples of different sanding processes and 6-gingerol simulated processing products were determined,and the correlation between 6-gingerol content and antioxidant activities was analyzed by SPSS 23.0.2.To investigate the mechanism of 6-gingerol and its transformed products in improving vascular endothelial injury based on network pharmacologyThe component targets of transformation product 6-shogaol,zingerone and prototype component 6-gingerol were obtained through Swiss Target Prediction database and Pharm Mapper database,and the vascular endothelial injury targets were obtained through Gene Cards database.Venn website was used to screen the intersection targets of the three component targets and vascular endothelial injury targets.The PPI relationship network was constructed for the intersection targets through STRING online database.GO functional enrichment and KEGG pathway analysis were performed for the intersection targets through DAVID data.Cytoscape 3.8.0 software was used to construct the "component-target" interaction network diagram,PPI interaction network diagram and "component-target-pathway" interaction network diagram for topological analysis.3.6-gingerol and its transformed products regulate ferroptosis through PI3K-AKT-NRF2 pathway to improve LPS-induced HUVEC damageThe experiment was divided into normal group,LPS model group,LPS+ 6-shogaol group,LPS+ Zingeron group and LPS+ 6-gingerol group.Cell viability was detected by CCK8 method,cell apoptosis was observed by Hoechst 33258 staining,and the expression levels of IL-1β,TNF-α,ROS,MDA,SOD,GSH and Iron were determined by ELISA and Ferrozine microplate method.The expression levels of HUVEC-related genes in each group were detected by q RT-PCR,and the expression of HUVEC-related proteins was detected by Western Blot.Result:1.Study on the transformation rule of 6-gingerol and the change of antioxidant activity in the processing of processed gingerThe contents of 6-shogaol and Zingerone in the samples increased gradually,and the contents of 6-gingerol decreased gradually with the extension of sanding time and the increase of temperature.The simulated processing confirmed that 6-gingerol can be partially converted into 6-gingerol and Zingerone after heating in oil bath.There was a significant positive correlation between the content of 6-gingerol and the antioxidant activity in samples of processed ginger and 6-gingerol simulated processing.2 To investigate the protective effect of 6-gingerol and its transformed products on vascular endothelial injury based on network pharmacologyA total of 255 targets of 3 components,986 targets of vascular endothelial injury and172 targets of intersection were obtained through screening of relevant databases.Topological analysis of Cytoscape 3.8.0 selected 10 key targets including SRC,PIK3R1,PIK3 CA,GRB2 and HSP90AA1.The results of GO enrichment analysis and KEGG enrichment pathway showed that the endothelial protection of the transformation products6-shogaol and Zingerone and the prototype component 6-gingerol mainly involved the positive regulation of cell migration,the negative regulation of apoptosis,the positive regulation of phosphatidylinositol 3-kinase signal transduction,and the positive regulation of cell proliferation.three components on the PI3K-AKT signaling pathway,regulating its downstream target genes and target proteins.Based on literature analysis,it is speculated that the protective effects of 6-shogaol and Zingerone and 6-gingerol on vascular endothelial may be related to the regulation of ferroptosis through the PI3 KAKT signaling pathway and the downstream NRF2/HO-1 pathway.3 6-gingerol and its transformed products regulate ferroptosis through PI3K-AKT-NRF2 pathway to improve LPS-induced HUVEC damageThe results showed that compared with the normal group,the cell viability was significantly decreased,the cell morphology was changed,and the nucleus was broken,while the cell viability was significantly increased and the cell morphology was significantly improved in the groups treated with LPS+ 6-shogaol,LPS+ Zingerone and LPS+ 6-gingerol.ELISA results showed that LPS model group could increase the expression levels of IL-1β,TNF-α,ROS,MDA,Iron,and reduce the expression levels of SOD and GSH.The results of q RT-PCR showed that LPS model group could up-regulate the expression levels of PI3 K,AKT,NRF2 and other pathway genes,up-regulate the expression ratio of Bax/Bcl-2 gene that promotes apoptosis,and down-regulate the expression levels of GPX4 and SLC7A11 genes that negatively regulate ferroptosis.Western Blot results showed that there were no significant differences in the expression levels of PI3 K and AKT among all groups,but LPS model group could significantly reduce p-PI3 K and p-AKT,and the other indexes were consistent with the results of q RTPCR.LPS+ 6-shogaol,LPS+ Zingerone and LPS+ 6-gingerol could reverse the above trend.Conclusion:1.Through the method of simulating processing combined with in vitro antioxidant,it is clear that 6-gingerol will be converted into 6-shogaol and Zingerone during the processing of processed ginger.With the extension of sanding time and the increase of temperature,the contents of 6-shogaol and Zingerone gradually increased,while the content of 6-gingerol gradually decreased,and there was a significant positive correlation between 6-gingerol content and antioxidant activity.2.On the basis of literature analysis,through network pharmacology combined with in vitro cell verification,it was confirmed that the transformation products 6-shogaol and Zingerone and the prototype component 6-gingerol inhibited ferroptosis by activating the PI3K-AKT-NRF2 pathway to improve HUVEC oxidative stress injury induced by LPS. |
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