| Purpose:By detecting the changes of IL-17,TGF-β,Foxp3,RORγt and total Ig E in the skin tissue of the sensitized part of the back of urticaria mice,the mechanism of regulating Th17 /Treg related transcription factors in urticaria mice by acupuncturing Quchi,Xuehai and Zusanli points was studied,so as to provide a new basis for the clinical treatment of urticaria.Material and method:In this experiment,40 ICR male mice aged 8 weeks,weighing20-22 g,were adaptively fed for one week.They were grouped by random number table method,which were divided into normal group,model group,acupuncture group and positive drug group,10 mice per group.The experiment adopted the pretreatment method of treatment before modeling.In the acupuncture group,mice were selected for acupuncture at Quchi,Xuehai and Zusanli points;the positive drug control group was given loratadine 0.2mg,and the two groups were treated for 7 days.On the 8th day,a passive skin allergic reaction model was established: the mice were anesthetized with ether,and the skin in the middle of the back of each group was sheared.Except for the blank group,the other 3 groups were injected subcutaneously with 0.5% anti-DNP-Ig E phosphate near the lumbar spine.The saline solution was 0.1 ml,and the blank group was intradermally injected with 0.1 ml of 0.9% sodium chloride solution at the same position.After 48 hours,the four groups were challenged with antigen,and the tail vein was injected with 200 μl of 0.9% phosphate solution containing DNP-HSA 0.5 μg / μl + 1% Evans Blue.After 2 hours,the abdominal arterial blood of the mice and the skin of the back sensitization site were taken.Hematoxylin-eosin(HE)staining and enzyme-linked immunosorbent assay were used to observe the total serum Ig E levels in mice.Immunohistochemical methods were used to observe the IL-17,TGF-beta,and transcription factors Foxp3,The effect of RORγt protein expression.The expression levels of Foxp3 and RORγt protein in skin tissues of mice were observed by Western blot.Results:1.Results of total serum Ig E levels of mice in each group:Compared with the normal control group,the total serum Ig E level of the model controlgroup was significantly higher,the difference was statistically significant(P < 0.05);although the total serum Ig E level of the acupuncture group and the positive drug group did not return to the normal value,it was significantly lower than that of the model control group(P < 0.05);compared with the acupuncture group,the total serum Ig E level of the positive drug group was significantly lower,the difference was statistically significant(P < 0.05).2.HE staining results of skin tissue of mice in each group:The epidermal layer is thick and normal,and its cells have a complete and uniform arrangement,and the structure is revealed without abnormality in normal group.: The epidermal layer is significantly thickened,there is inkeratosis or excessive keratosis,the dermis is swollen and deformed,and the inflammatory in model group.The epidermis is slightly thickened,the keratosis is mild,the dermis is slightly edema,and the inflammatory cells are replaced See,the blood vessels are slightly dilated in positive drug group.:The skin and epidermis are slightly thickened,the structure is basically intact,the edema of each layer is not obvious,and occasionally there are a few inflammatory cells in acupuncture group.3.Immunohistochemical results of cytokines IL-17,TGF-β,Foxp3,and RORγt in the skin tissue of mice in each group:The expression of TGF-β and FOXP3 in the model group was significantly lower than that in the blank group(P <0.05);In the acupuncture group,the positive drug group significantly increased the expression of TGF-β and FOXP3-positive cells in the skin tissue of the sensitized area of the mouse instead of the model(P <0.05).Compared with the acupuncture group,the positive drug group TGF-β,FOXP3 and positive cells were slightly increased.Compared with the blank control group,the model group have increased the expression of IL-17 and RORγt-positive cells in the skin tissue of the mouse sensitization site(P<0.05).Compared with the model mice,the expression of the acupuncture group and the positive drug group of IL-17 and RORγt positive cells in the skin tissue of the sensitized area was significantly reduced(P <0.05).Compared with the acupuncture group,the IL-17,RORγt and The expression of cells was slightly reduced.4.The expression of RORγt and Foxp3 protein in the skin tissue of mice:Compared with the normal control group,the expression of RORγt protein in the model control group was significantly increased(P < 0.05);compared with the model control group,the expression of RORγt protein in the acupuncture group and the positive drug group was significantly reduced(P < 0.05);compared with the acupuncture group,the expression of RORγt protein in the positive drug group was significantly reduced(P < 0.05).Compared with the normal control group,the expression of Foxp3 protein in the model control group was significantly reduced(P < 0.05);compared with the model control group,the expression of Foxp3 protein in the acupuncture group and the positive drug group increased significantly(P < 0.05);compared with the acupuncture group,the expression of Foxp3 protein in the positive drug group increased significantly(P< 0.05).Conclusion:1.Acupuncture can inhibit type I hypersensitivity by reducing the total serum Ig E level and improving the morphological and pathological changes of the skin of urticaria mice.2.Acupuncture can promote Foxp3 protein expression and TGF-β production,inhibit RORγt protein expression and IL-17 production,promote Treg cell differentiation and inhibit Th17 cell differentiation,restore the balance between Foxp3 and RORγt,so as to regulate Th17 /Treg balance axis,reduce the skin inflammation reaction of sensitized site,which may be the mechanism of acupuncture to improve urticaria symptoms. |
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