Exploration Of The Mechanism Of Action Of Hooping Ointment In Treating Infected Diabetic Ulcer Based On Transcriptomic

Objective:Observation of the therapeutic effect of Guwei ointment in the treatment of infected diabetic ulcers and the exploration of its mechanism based on the transcriptomics technology.Method:1 Construct the non-infectious ulcer model,infectious ulcer model and infectious diabetic ulcer model.Compare the wound healing rate and HE staining results of the tissues around the wound of the three groups to verify whether the model achieves the expected effect.2 Establish a model of infected wounds in diabetic rats,and treat the wounds with iodine-impregnated gauze or Guwei ointment according to the groups.Measure changes in wound area and observe morphological changes in wound tissues by pathological section analysis to validate the efficacy of Guwei ointment for the treatment of infected diabetic ulcers.3 Use transcriptome sequencing technology to screen for differentially expressed genes(DEGs)based on the standard of |log2FoldChange|>1 and significant P-value<0.05.Candidate genes and pathways that may play a role in the anti-inflammatory and pro-healing effects of Guwei ointment in treating infected diabetic ulcers were screened using GO enrichment analysis and KEGG pathway enrichment analysis.Result:1 The wound healing rate of infectious ulcer model was lower than that of non-diabetic ulcer model,and the healing rate of infectious wound in diabetic rats was lower than that of non-diabetic rats.2 Efficacy observation:The healing rate of the control group,treatment group were all faster than that of the model group.Among them,the healing rate of treatment group was the fastest.On the 7th day,the healing rate of treatment group was superior to that of the control group(P<0.05),and the difference was statistically significant.2 HE staining:Before and after intervention,the number of inflammatory cells in the dermal papilla tissue of each group decreased compared to before,and newly formed capillaries were visible.Compared with the control group,there were more newly formed capillaries and increased follicular glandular tissue in treatment group and self-contrast group.3 Masson staining:Before and after intervention,scattered collagen fibers with disordered arrangement were visible in all groups’ fields of view.Compared with the control group,the collagen fiber bundles within the field of view were more closely arranged in treatment group and self-contrast group after treatment,4 Transcriptomics:155 DEGs were identified between the control group and treatment group,2887 DEGs were identified before and after intervention in the control group,and 3683 DEGs were identified before and after intervention in self-contrast group.The GO enrichment results show that,compared to the control group,in treatment group,the DEGs are mainly enriched in keratinization,oxidoreductase activity,and pigment formation processes.When comparing pre-and post-intervention in the control group,the DEGs are mainly enriched in intermediate filaments,protein binding,and tissue development processes.When comparing pre-and post-intervention in self-contrast group,the DEGs are mainly enriched in extracellular region,protein binding,and cellular regulation processes.The KEGG pathway annotation classification and enrichment results showed that DEGs between the control group and treatment group were mainly enriched in tyrosine metabolism,melanogenesis,and NF-κB signaling pathway.The DEGs before and after intervention in the control group were mainly enriched in axon guidance pathway,Hippo signaling pathway,and DNA replication pathway.The DEGs before and after intervention in self-contrast group were mainly enriched in chemotaxis factor.Conclusion:1 The wound healing of the infected ulcer model was delayed than that of the non-infected ulcer,and the infected wound healing of the diabetic rats was delayed than that of the non-diabetic rats.The experimental modeling was successful and the model was stable.2 The application of Guwei ointment could reduce the infiltration of inflammatory cells in the wound,promote neovascularization,adjust the arrangement of collagen fibers,thereby promoting the healing of infectious diabetic ulcers,and improving the wound healing rate in the later stage of wound repair.3 The effective promotion of healing for infectious diabetic ulcers by Guwei ointment may be related to the regulation of NF-κB signaling pathway and Cytokine-cytokine receptor interaction pathway.The possible targets are Edar gene and CCL2 gene.