| Objectives:In this study,different types of macrophages(M1,M2)were obtained by mononuclear cell differentiation-induced differentiation technique.Mycoblasts were induced by different concentrations of curcumin.The macrophages were induced by macrophages.The effects of different concentrations of M1 macrophages induced by different concentrations on wound healing were discussed by several factors of healing and cytokine determination.Methods: Extraction and activation of bone marrow mononuclear cells.The rat bone marrow cells were extracted and purified by differential adherence and M-CSF stimulation.The macrophages required for the experiment were activated and the cell results were analyzed by flow cytometry.Macrophage-induced differentiation.IL-4 stimulated the transformation of macrophages M0 to M2,and demonstrated the effectiveness of M1 and M2 induced by flow cytometry analysis.M1-type macrophages in the re-induced differentiation.Different concentrations of curcumin stimulated M1 macrophages to transform M2 type macrophages to obtain group B cell suspension.The mice were treated with M1 and M2 macrophages with the corresponding concentration of solvent(DMSO + PBS)to obtain group A and C group of cell suspension;and the use of flow cytometry analysis of curcumin M1 on the induced polarization efficiency.Preparation of rat trauma model and evaluation of its usability.Six models of 6mm diameter skin penetrating injury model were made on the back of the experimental rats.The availability of the model was evaluated by HE staining analysis.The experimental conditions of the wound were evaluated and evaluated.The experimental animals were divided into 5 groups: group 1,group 3,group 7,group 10 and group According to the principle of randomized double-blind,the six wounds of each rat were treated as follows:(1)Group A cell suspension was injected in the local(subcutaneous,dermis and basal infiltration).(2)do not do any treatment.(3)local injection of group B cell suspension.(4)local injection sterilization PBS.(5)local injection of group C cell suspension.After the number of experimental days,the rats were sacrificed and the specimens were tested and analyzed.The differences in healing rates were observed by macroscopic analysis of HE staining,MASSON staining and immunohistochemical analysis.Results: 1.The ratio of CD14 and CD68 double positive cells in rat bone marrow cells was up to 86.5%.2.Curcumin stimulus M1 macrophages to M2 macrophage polarization role the best concentration of 25 umol/L,polarization efficiency is 29.59%.3.There were no significant differences in the wound healing rates of the six different anatomical sites in the back of the rats.There was no significant difference in HE staining results.Compared with the blank control group,the wound healing delay was observed in the M1 + Cur0 group.HE staining and MASSON staining showed that the infiltration rate of inflammatory cells was long and the granulation tissue grew slowly.The wound healing rate,VEGF,b FGF,B1)quantitative analysis suggested that the relevant indicators than the blank control group there is a statistical difference.M2 + Cur0 group and M1 + Cur25 group were observed before healing,the results of the above indicators and M1 + Cur0 group and the existence of statistical significance.There was no significant difference between PBS group and blank control group.Conclusions 1.M-CSF can induce rat bone marrow cells to form macrophages in vitro.2.LPS and IL-4 induce the transformation of macrophages to M1 and M2,respectively.3.Curcumin can induce M1 type macrophages to M2 type polarization,and the best polarization efficiency of 29.59%.4.A method of preparing a rat wound healing model by punching method is available.5.Curcumin regulates typing-mediated macrophages to promote wound healing. |
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