Influence Of MEBT/MEBO On Expression Levels Of NF-κB P65, IκBα, IKK And Their Phosphorylated Proteins In Chronic Refractory Wounds

Objective:To investigate the influence of the regenerative medical technology for burns,wounds and ulcers(Moist Exposed Bum Therapy/Moist Exposed Burn Ointment,MEBT/MEBO)on expression levels of nuclear factor-κB(NF-κB)p65,inhibitor-κbinding protein-nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor,alpha(IκBα),IκB kinase(IKK)and their phosphorylated proteins(p-NF-κB p65,p-IκBα,P-IKK)in chronic refractory wounds(Chronic Refractory Wounds,CRW).Methods:Ninety SPF-bred male Wistar rats were selected and,after adaptive feeding for one week,randomized into five groups:blank group,control group,model group,MEBO group and rb-b FGF group,18 rats in each group.Only skin preparation was performed on rats in blank group,whereas acute wound models were established on rats in control group,and chronic refractory wound models were made on rats respectively in model group,MEBO group and rb-b FGF group.After model establishment,the prepared skin in blank group and the wounds in control group and model group were managed with dressing change of normal saline gauzes,and the wounds in rb-b FGF group and MEBO group were respectively managed with dressing change of recombinant bovine basic fi-broblast growth factor(rb-b FGF)and MEBO.The wound healing rate,histopathological changes and the expression levels of NF-κB p65,IκBα,IKK and p-NF-κB p65,p-IκBα,p-IKK in the skin or wound tissues of rats in each group were com-pared on day 3,7 and 14 of intervention.The fluorescence intensity of NF-κB p65 was measured by immunofluorescence,and the expression trend of inflammatory cytokines was observed indirectly andsemi-quantitatively.Results:(1)No significant difference was observed among the five groups on day 3 and 7 of intervention intermsofwound healing rate(allp>0.05);onday14ofintervention,thewoundhealing rate in model group was significantly lower than that in control group and MEBO group(both p<0.05),while no significant difference in wound healing rate among other groups(all p>0.05).The wound healing time of MEBO group was not significantly different from that of control group and Beifuxin group(p > 0.05).Compared with model group,the healing time of MEBO group was significantly shorterthanthat ofmodelgroup(p < 0.05),andthe differencewasstatistically significant.The wound healing time of other groups was also significantly different(p < 0.05),and the healing timewas shorterthanthatofmodel group.(2)Onday7 ofintervention,theamount ofcollagen fibers in the wound tissues of rats increased significantly,and a large number of fibroblasts,newly born capillariesandfewhairfolliclestructureweregeneratedin MEBOgroup andrb-b FGFgroup;on day14 of intervention,the numbers of collagen fibers in the wound tissues of rats in MEBO group and rb-b FGF group were obviously more than that in model group,with orderly arranged capillaries,mature hair follicles and sebaceous glands visible;In model group,infiltration of a small num-berofinflammatory cells was still observableinthewound tissues,buttherewerealready a lotof fibroblasts generated.(3)On day 7 and 14 of intervention,the expression levels of NF-κB p65,IKK,p-NF-κB p65,p-IKK and p-IκBαin the skin or wound tissues of rats in blank group,control group and MEBO group were significantly lower than that in model group(all p<0.05);on day 7 of intervention,the expression levels of NF-κB p65 and IκBαin the wound tissues of rats in MEBO group were significantly higher than that in rb-b FGF group(both p<0.05),whereas the expression levels of IKK,p-NF-κB p65,p-IKK and p-IκBαwere significantly lower than that in rb-b FGF group(all p<0.05);on day 14 of interven-tion,the expression levels of IKK and IκBαin the wound tissues of rats in MEBO group were obviously lower than that in rb-b FGF group(both p<0.05).(4)On day 3 of modeling,the immunofluorescence intensity of NF-κB p65 in control group,Beifuxin group,model group and MEBO group was compared with that in blank group,p < 0.05,indicating statisticalsignificance;On day 7,the fluorescenceintensity ofcontrol group,model group and MEBO group was statistically significant compared with blank group(p < 0.05).On the 14 th day,the control group,Beifuxin group,model group and MEBO group had statistical significance compared with blank group(p < 0.05),among which,MEBO group had statistical significance with Beifuxin group,modelgroupand blankgroup(p < 0.05).Conclusion:The mechanism of MEBT / MEBO in promoting the healing of chronic refrac-tory wounds may be associated with the expression levels of NF-κB p65,IκBα,IKK and their phosphorylated proteinsinwoundtissues.