| Background:Keloid is the result of trauma or injury. In some cases, the keloid grows out ofcontrol.Keloid, especially locats in the cosmetic sensitive area causes severepsychological and dysfunctional probems of the patient.Keloid is a kind of pathologicalscar. It is a common disease in plastic surgery, while its etiology is not clearlydemonstrated. Although clinical therapies for keloid are developing, high recurrence isstill a problem. An efficient treatment for keloid is badly needed.The mechanism under the hyperplasia of keloid is not clear. Fibriblast is the maineffector cell which secrets the extracellular matrix.The source of the modulator whichpromote the fibrosis of the fibroblast is unknown. The immunocell may be an importantsource of fibrosis factors. Tacrolimus is an anti-immunological medicine belonging tomacrolide. It is proved to inhibit the activity of lymphocytes. Tacrolimus is used to treatatopic dermatitis which is negative to hormone therapy. It is found that keloid of thepatients was released at the mean time. We proved that tacrolimus inhibit the hyperplasiaof hypertrophic scar in rabbit model. In this study, we demonstrated the function ofimmunology cells in keloid formation. Mean-while, we are exploring the mechanism that tacrolimus cure keloid by inhibitingthe immunology cells. Therefore we can blaze a newtrail in keloid treatment and prevetion.Aim:Establish a co-culture model of keloid fibroblast and PBMCs. Exploring themechanism tacrolimus inhibits PBMCs from secreting fibrotic factors. Demonstratingthe inhibition of hyperplasia and collegen secretion, then find a new way of keloidtheatment and prevetion.Method:1. Set up a co-culture system of keloid and fibroblast with PBMCs. We detectedCollagen I with real-time PCR. Then we check the proliferation of fibroblast after24hrs PBMCs supernatant treatment by MTT Assay.2. The same assay and treatmen were used in PBMCs co-culture system. Then real-timePCR and western blot were used to detect College I.3. Detect the related fibrosis factors expression of PBMCs in upper chamber of theco-culture system. MTT Assay detected the proliferation of keloid fibroblast thattreated with Tacrolimus in different dosage.4. Explore the keloid fibroblasts feedback regulation of PBMC with real-timePCRdetection the expression of IL–4、IL-6、IL-10.Result:1. In co-culture system, the expression of Collegen I increased in normal fibroblast andkeloid. The keloid is more sensitive to PBMCs. Keloid fibroblast grows faster than thenormal group. PBMCs are more powerful to keloid fibroblast.2. Co-culture system, keloid fibroblast secreted significantly high level of Collegen I andCollegen III, P＜0.01. Collegen I and III as well as the inflammatory factorsexpression were suppressed by Tacrolimus, P＜0.01. The effect of Tacrolimus in20ng/ml group is significantly, P＜0.01. 3. Tacrolimus can significantly inhibit the expression of inflammatory cytokines,20ng/ml group display significantly effect (P <0.01)，Tacrolimus made no differencewhen it is solely given.After adding PBMCs supernatant, the cell proliferationincreased. In20ng and100ng group proliferation were inhibited by Tacrolimus.4. Keloid fibroblasts culture supernatant had stronger stimulation to PBMC, comparedwith normal skin or hyperplastic scar fibroblasts supernatant.Conclusion:We established a co-culture system of fibroblasts and PBMCs. The experimentsimulates the effect of PBMCs on keloid fibroblast. We proved that keloid is moresensitive to this affect that normal foborblast. Meanwhile Tacrolimus has no solo functionon fibroblast. Therefore Tacrolimus may cure keloid through the way that inhibit thesecretion function of immunology cells... |
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