Molecular Pathway Mechanism And Clinical Research Of Bombyx Mori In The Treatment Of Scars Based On Network Pharmacology

Purpose:Currently,there is a lack of research on the mechanism of the treatment of cicatricial diseases by Bombyx batryticatus(BB).The target genes,compounds and KEGG signaling pathway of the BB in the treatment of cicatricial diseases are studied by network pharmacology,bioinformatics analysis and molecular targeted docking,to reveal the molecular pathway and mechanism of the BB in the treatment of keloid or hypertrophic scar.Further,the silkworm(0.5% menthol as the transdermal penetrant matrix)is used for local dressing change of postoperative wound in patients with anal fistula,in order to observe the clinical effect of the treatment of postoperative scar.Material and method:1.Network pharmacology techniques were fully adopted to effectively predict the active ingredients of the BB,which were then screened to obtain the common disease-drug target genes.This study then further investigated the molecular targets,and KEGG signaling pathways of Bombyx batryticatus(BB)in the treatment of keloid.To construct the interaction network of the BB's main drug components,target genes and molecular signaling pathways for the treatment of keloid.In addition to the CCK8 cell proliferation inhibition assay and Transwell cell invasion assay,in-vitro cell experiments were conducted to validate the research findings.The expression levels of MMP-1,MMP-9,Akt,P-Akt,beta-Actin,P13 K p85?,p38 MAPK,GAPDH,and NF-?B in human hypertrophic scar fibroblasts(h HSFs)were evaluated after intervention with different experimental groups(the control group;BB groups with different concentrations;beauvericin group;and a PI3K/Akt inhibitor LY294002),respectively.In addition,three main chemical constituents(beauvericin,cyclo(D)-Pro-(D)-Phe,and(+)-pinoresinol)and four key scar intervention targets(MMP1,MMP9,MMP13,and PI3k)were used as the research objects.Using molecular targeting docking,the interaction between the three main chemical components of BB and disease targets was further discussed.2.Seventy-six patients with anal fistula hospitalized in the anorectal department of Xianyang Central Hospital from February 2020 to August 2020 were collected,with 38 patients in each group.The observation group was given local topical use of Bombyx batryticatus(0.5%menthol as the matrix of transdermal penetration agent)after surgery,while the control group was given local topical use of Triamcinolone acetonide after surgery.During the study,a random number table was used and randomly assigned to the observation group and the control group.The baseline characteristics,the reduction degree of Vancouver Scar Scale(VSS),the width of Scar(ultrasound)and other indicators of the two groups were analyzed.H&E staining was used to compare the histological morphology of two groups before and after intervention.Immunohistochemical staining was used to detect the number of vascular endothelial growth factor(VEGF),proliferating cell nuclear antigen(PCNA)and Vimentin positive cells in the wound tissue of the two groups before and after intervention.Postoperative complications(pigmentation,vascular distribution,flexibility,scar height,skin atrophy,telangiectasia,local ulcer,hypopigmentation)indexes of the two groups were observed,recorded and analyzed.The clinical efficacy,wound healing time,wound healing rate,satisfaction degree and other data of the two groups were analyzed,and the clinical efficacy evaluation of Bombyx batryticatus(0.5% menthol as the matrix of transdermal penetration agent)on inhibiting scar formation after anal fistula operation was made.Results:1.After screening out 73 active components related to scar disease and 148 molecular signal pathways out of 688 target genes of BB,the interaction network of the BB's main drug components,target genes and molecular signaling pathways was constructed.It was found that BB played a key role in anti-oxidation,apoptosis,complement activation,inflammatory response,cell migration,cell matrix reconstruction,anti-tumor,improvement of hypoxia,and senescence regulation.In vitro experiments showed that compared with the blank control group,the expression of MMP-1,MMP-9,p-Akt,p38 MAPK and NF-?B in proliferating scar fibroblasts were significantly decreased in BB group,Beauvericin group and LY294002group(P<0.05).The PI3K/Akt/MMP1/MMP9 signaling pathway could be effectively inhibited under the intervention of BB's crude extract and Beauvericin.Based on the Autodock Vina 1.1.2 docking score,the calculation results indicated that 3 compounds(the Beauvericin,Cyclo(D)-Pro-(D)-Phe and(+)-Pinoresinol)may simultaneously act on four targets(MMP1,MMP9,MMP13,and PI3k).That are suggesting that BB could exert synergistic effects to inhibit the PI3K/Akt/MMP1/MMP9 molecular signaling pathway,by acting on multiple targets.2.Among the 76 patients with anal fistula,the wound healing time of the treatment group treated with BB(0.5% menthol as transdermal enhancers penetrant matrix)was better than that of the triamcinolone acetonide control group(P<0.05).The wound healing time of the BB group was 25.39±3.03,and the wound healing time of the triamcinolone acetonide control group was 29.48±2.90,respectively.The difference in wound healing time and wound healing rate between the two groups was statistically significant(P<0.05).After intervention between the treatment group and the control group,there were significant differences in the reduction degree of the Vancouver Scar Scale(VSS)and the width of scar in ultrasound examination(P<0.05),and there were statistically significant differences in the staining number of vascular endothelial growth factor(VEGF),proliferating nuclear antigen(PCNA)and Vimentin positive cells in the tissues of the two groups(P<0.05).The postoperative complications(flexibility,scar height,skin atrophy,telangiectasia,hypopigmentation)were significantly different between the two groups(P<0.05).The BB(0.5% menthol as permeating agent matrix)was beneficial to reduce the formation of scar after anal fistula(P<0.05)and reduce postoperative pain.There was no significant difference in pigmentation between the two groups after anal fistula operation(P>0.05).The treatment group does not appear local and systemic adverse reactions.Conclusion:1.Bombyx batryticatus can effectively inhibit the PI3K/Akt/MMP1/MMP9 molecular signaling pathway,and can also effectively inhibit the proliferation and invasion of human hyperplastic scar fibroblasts,and regulate the occurrence and development of scar diseases.2.In the wound healing after anal fistula operation,the BB(0.5% menthol as permeable agent matrix)has the advantages of simple operation,small trauma,less pain and good curative effect,which is conducive to reducing the formation of postoperative scar after anal fistula,which is worthy of clinical development.