| Purpose:Using two nitrochlorobenzene(DNCB)mice ACD model is set up,by ACD changes in areas of organization form,pathological changes in mice,and established evaluation based on the principle of tomifying qi and blood(needle medicine with method of inhibition of ACD,through to the ACD model related cytokines in mice and its mRNA level determination,from the cellular and molecular level to explore its mechanism of action,in order to better use needle medicine and clinical treatment of ACD provide experimental support.Material and method: 1 Material There were 60 white male mice in the SPF grade kunming,4 weeks old,and the weight was(20±2)g.2 Method 2.1 Building methods The ACD mouse model was established with a small dose of DNCB solution.2.2 Observation of the changes of ear skin lesions in ACD mice.The inhibition effect of ACD on ACD was preliminarily evaluated by observing the difference in thickness and weight of the left and right ear of ACD mice.2.3The morphological changes of skin lesions in ACD mice were observed.By observing the morphological and pathological changes of the skin lesions in ACD mice,the effects of acupuncture on specific acupoints and yiqi decoction and the allergic reaction of ACD mice were discussed.2.4 Changes of il-6,il-10 and il-17 in serum of mice were detected by Elisa.With Elisa method to detect after acupuncture with yiqi functions of soup and ACD in mice serum il-6,il-10,the levels of il-17,interpretation of acupuncture and yiqi functions of soup and the adjustment role of cytokines in ACD mice blood,then explain the ACD mice a balance of Th17/Treg cells in the blood of the influence of ACD.2.5 RT-PCR was used to detect the expression of il-17 mRNA and Foxp3 mRNA in mouse pathological tissues.By rt-pcr method to detect acupuncture with yiqi functions of soup and ACD il-17 in the diseased tissue of mice after the expression of mRNA,Foxp3 mRNA,acupuncture and yiqi functions of soup with the ACD related factor mRNA content in diseased tissue in mice,mice that ACD of Th17 and Treg cells in diseased tissue changes.2.6 Western Blot was used to verify the expression of il-17 mRNA and Foxp3 mRNA in mouse pathological tissues.Using Western Blot method to detect acupuncture with yiqi functions of soup and ACD IL-17 in the diseased tissue of mice after the expression of mRNA,Foxp3 mRNA,from another Angle shows that acupuncture with yiqi functions of soup and ACD of relevant factors in the diseased tissue in mice after mRNA content change,further illustrate the influence of the balance of ACD of Th17/Treg cells.Results: 1.Changes of ear skin lesions in ACD mice.1.1 Variation of ear thickness of each group,the difference between the model group,acupuncture group,Chinese medicine group,acupuncture group and western medicine group was significantly higher than that in the blank group(P < 0.05).Comparison of acupuncture group,traditional Chinese medicine group,acupuncture group,western medicine group and model group showed significant decrease in ear thickness(P < 0.05).Compared with the acupuncture group,there was no difference in the Chinese medicine group(P>0.05),while the thickness of the ear between the acupuncture group and the western medicine group was lower than that in the acupuncture group(P < 0.05).Compared with the traditional Chinese medicine group,the ear thickness difference between the acupuncture group and the western medicine group was lower(P < 0.05).Compared with the acupuncture group,there was no significant difference in the western medicine group(P>0.05).1.2The weight difference of each group was significantly higher than that in the blank group(P < 0.05),and the weight difference between the model group,acupuncture group,traditional Chinese medicine group,needle medicine group and western medicine group was significantly higher than that in the blank group(P < 0.05).Comparison of acupuncture group,traditional Chinese medicine group,acupuncture group,western medicine group and model group showed significant decrease in ear weight difference(P < 0.05).Compared with the acupuncture group,there was no difference in the Chinese medicine group(P BBB 0 0.05),while the ear weight difference between the acupuncture group and the western medicine group was lower than that in the acupuncture group(P < 0.05).Compared with the traditional Chinese medicine group,the ear weight difference between the acupuncture group and the western medicine group was lower(P < 0.05).Compared with the acupuncture group,there was no significant difference in the western medicine group(P>0.05).Pathological changes of skin lesions in ACD mice.2.1The normal structure of epidermal thickness was observed in the blank group,and the cells were closely aligned,and there was no intracellular and intracellular edema,and occasionally mild inflammatory cells were observed.2.2Model group observation skin thickening,excessive diversification,missing parts,intracellular and intercellular edema,increased cell spacing,swelling of the endothelial cells and dermal papilla edema,leather with a large number of inflammatory cells infiltration,main single nuclear cell of lymphocyte,rare polymorphonuclear cells and collagen fiber gap increases vasodilation.Observations of the skin structure basic complete 2.3Acupuncture group,skin mild parakeratosis,skin thickening,slightly mild edema between cells,cells,dermal papilla edema is not obvious,is one of the few inflammatory cells infiltration,mild swelling of the endothelial cells,damage degree of each layer model group significantly decrease relatively.2.4 Chinese medicine set of observations skin with mild parakeratosis,occasional edema cells and dermal papilla mild edema,there are a few inflammatory cells infiltration,mild swelling of the endothelial cells,each layer damage than model group significantly reduce.The results showed that the epidermal thickness was approximately normal,and there was mild angularity,and there were a few inflammatory cells infiltrating in the dermis.The vascular endothelial cells were mildly swollen,and the damage was significantly less than that of the model group.2.6 observations about normal epidermis thickness,western medicine group,occasional mild parakeratosis,occasional edema cell,leather inside with a small amount of inflammatory cells infiltration,the layers of damage significantly lighter than model group,and needle medicine group similar to skin structure.3 Elisa method was used to detect the results of il-6,il-10 and il-17 in serum of mice.3.1 ELISA method was used to detect il-6 in peripheral blood of each group of mice,and the expression of il-6 in peripheral blood of the model group,acupuncture group,Chinese medicine group,acupuncture group and western medicine group was significantly higher than that in the blank group(P < 0.01).The expression of il-6 in peripheral blood was significantly decreased in acupuncture group,traditional Chinese medicine group,acupuncture group,western medicine group and model group(P < 0.01).Compared with the acupuncture group,there was no difference in the TCM group(P>0.05),while the expression of il-6 in the peripheral blood of the acupuncture group and the western medicine group was lower than that in the acupuncture group(P < 0.05).Compared with the traditional Chinese medicine group,the expression of il-6 in the peripheral blood of the acupuncture group and the western medicine group was significantly reduced(P < 0.01).Compared with the injection group,there was no significant difference in the western medicine group(P>0.05).3.2 ELISA method was used to detect il-10 in peripheral blood of each group of mice,and the expression of il-10 in peripheral blood of the model group,acupuncture group,Chinese medicine group,acupuncture group and western medicine group was significantly lower than that in the blank group(P < 0.01).The expression of il-10 in peripheral blood was significantly increased in acupuncture group,traditional Chinese medicine group,needle medicine group,western medicine group and model group(P < 0.01).Compared with the acupuncture group,there was no difference in the Chinese medicine group(P>0.05),while the expression of il-10 in the peripheral blood of the acupuncture group and the western medicine group was lower than that in the acupuncture group(P < 0.01).Compared with the traditional Chinese medicine group,the expression of il-10 in the peripheral blood of the acupuncture group and the western medicine group was significantly reduced(P < 0.01).Compared with the injection group,there was no significant difference in the western medicine group(P>0.05).3.3The expression of il-17 in peripheral blood of mice in each group was detected by ELISA method,and the expression of il-17 in peripheral blood of the model group,acupuncture group,Chinese medicine group,acupuncture group and western medicine group was significantly higher than that in the blank group(P < 0.01).The expression of il-17 in peripheral blood was significantly decreased in acupuncture group,traditional Chinese medicine group,acupuncture group,western medicine group and model group(P < 0.01).Compared with the acupuncture group,there was no difference in the TCM group(P BBB 0 0.05),while the expression of il-17 in the peripheral blood of the acupuncture group and the western medicine group was lower than that in the acupuncture group(P < 0.01).Compared with the traditional Chinese medicine group,the expression of il-17 in the peripheral blood of the acupuncture group and the western medicine group was significantly reduced(P < 0.01),and there was no significant difference in the western medicine group compared with the acupuncture group(P>0.05).Expression of il-17 mRNA and Foxp3 mRNA in mouse pathological tissues.4.1RT-PCR was used to detect the expression of il-17 mRNA and Foxp3 mRNA in mouse pathological tissues.4.1.1 ACD il-17 mRNA expression in diseased tissue in mice,compared with the blank group,model group,acupuncture group,the Chinese medicine group,the needle medicine,western medicine group il-17 mRNA in a diseased tissue of mice increased significantly,obvious difference(P < 0.01);Compared with the model group,the expression of il-17 mRNA was significantly decreased in acupuncture group,traditional Chinese medicine group,needle medicine group,western medicine group and model group,and the difference was significant(P < 0.01).Compared with the acupuncture group,the expression of il-17 mRNA in the lesion tissues of the Chinese medicine group was not obvious(P > 0.05).Compared with the acupuncture group,il-17 mRNA in the lesion tissues of the acupuncture group and the western medicine group decreased significantly,and the difference was significant(P < 0.01).Compared with the traditional Chinese medicine group,the expression of il-17 mRNA was significantly decreased in the group of acupuncture and western medicine group,and the difference was significant(P < 0.05).There was no significant difference in the expression of il-17 mRNA in the needle and western medicine group(P>0.05).4.1.2 ACD Foxp3 mRNA expression in the diseased tissue in mice,compared with the blank group,model group,acupuncture group,the Chinese medicine group,the needle medicine,western medicine group mice Foxp3 mRNA decreased obviously in the diseased tissue,obvious difference(P < 0.01);Comparison of acupuncture group,traditional Chinese medicine group,needle medicine group,western medicine group and model group showed that Foxp3 mRNA increased significantly in the lesion tissues,with significant difference(P < 0.01).Compared with the acupuncture group,the expression of Foxp3 mRNA in the lesion tissues of Chinese medicine group was not obvious(P>0.05).Compared with the acupuncture group,the expression of Foxp3 mRNA in the lesion tissues of the acupuncture group and the western medicine group was significantly increased,and the difference was obvious(P < 0.01).Compared with Chinese traditional medicine group,acupuncture group Foxp3 mRNA expression in diseased tissue in mice increased significantly,obvious difference(P < 0.05),the western medicine group Foxp3 mRNA expression in diseased tissue in mice increased significantly,obvious difference(P < 0.01);Compared with the acupuncture group,there was no significant difference in Foxp3 mRNA expression in the lesion tissues of the western medicine group(P> 0.05).2 ACD il-17 mRNA expression in diseased tissue in mice,compared with the blank group,model group,acupuncture group,the Chinese medicine group,the needle medicine,western medicine group il-17 mRNA in a diseased tissue of mice increased significantly,obvious difference(P < 0.01);Compared with model group,acupuncture group,the needle medicine,western medicine group mice il-17 mRNA expression in diseased tissue decreased obviously,the significant difference(P < 0.01),Chinese medicine group of il-17 mRNA expression in diseased tissue in mice significantly decreased,the significant difference(P < 0.05);Compared with the acupuncture group,the expression of il-17 mRNA in the lesion tissues of the Chinese medicine group was not obvious(P > 0.05).Compared with the acupuncture group,il-17 mRNA in the lesion tissues of the acupuncture group and the western medicine group decreased significantly,and the difference was significant(P < 0.05).Compared with the traditional Chinese medicine group,the expression of il-17 mRNA was significantly decreased in the group of acupuncture and western medicine group,and the difference was obvious(P < 0.01).There was no significant difference in the expression of il-17 mRNA in the needle and western medicine group(P>0.05).4.2.2 ACD Foxp3 mRNA expression in the diseased tissue in mice,compared with the blank group,model group,acupuncture group,the Chinese medicine group,the needle medicine,western medicine group mice Foxp3 mRNA decreased obviously in the diseased tissue,obvious difference(P < 0.01);Comparison between acupuncture group,traditional Chinese medicine group and model group was not obvious(P >0.05).Compared with the model group,there was a significant increase in Foxp3 mRNA in the diseased tissue(P < 0.01).Compared with the acupuncture group,the expression of Foxp3 mRNA in the lesion tissues of the acupuncture group and the western medicine group was significantly increased,and the difference was obvious(P < 0.01).Compared with the traditional Chinese medicine group,the expression of Foxp3 mRNA in the lesion tissues of the acupuncture group and the western medicine group was significantly increased,and the difference was obvious(P < 0.01).Compared with the acupuncture group,there was no significant difference in Foxp3 mRNA expression in the lesion tissues of the western medicine group(P>0.05).Conclusion: 1.Changes of ear skin lesions in ACD mice.1.1 acupuncture and yiqi decoction can reduce the thickness and weight difference of the ear in ACD model mice,and the combination of the two is better.1.2 verify the loratadine suspension can decrease the thickness of the ACD model mice ear,weight difference,acupuncture with yiqi functions of soup and with loratadine suspension of ACD model mice ear thickness difference,almost equivalent weight difference repair function.2.Pathological effects of ACD mouse skin lesions.2.1 acupuncture,yiqi fusing decoction,acupuncture and yiqi decoction can reduce the pathological changes in ACD mice,but the effect of acupuncture and yiqi fusing soup is more obvious.2.2 verify the loratadine suspension can reduce the ACD model mice of diseased tissue pathological changes,acupuncture with yiqi functions of soup and with loratadine suspension of ACD model improvement of diseased tissue pathological changes in mice was similar,precise mechanism needs further research.The expression of il-6,il-10 and il-17 in serum of mice was detected by Elisa.3.1 acupuncture and traditional Chinese medicine can reduce the expression of il-6 and il-17 in peripheral blood,and increase the expression of il-10,and the effect of acupuncture is more obvious.3.2 verify the loratadine suspension can reduce the ACD model mice il-6,il-17 in the peripheral blood expression,higher expression of il-10,acupuncture with yiqi functions of soup with loratadine and to reduce the ACD model mice il-6,il-17 in the peripheral blood expression,higher expression of il-10,effect.Expression of il-17 mRNA and Foxp3 mRNA in mouse pathological tissues.4.1 the expression of il-17 mRNA and Foxp3 mRNA in the pathological tissues of ACD mice was detected by pt-pcr.4.1.1 acupuncture and yiqi decoction can reduce the expression of il-17 mRNA in the lesion tissues of ACD model mice,and increase the expression of Foxp3 mRNA,and the combination of the two is more obvious.4.1.2 confirmed that loratine suspension could reduce the expression of il-17 mRNA in the lesion tissues of ACD model mice,and increase the expression of Foxp3 mRNA,acupuncture and yiqi fusing decoction,and the effect similar to that of loratadine.4.2 expression of il-17 mRNA and Foxp3 mRNA in the pathological tissues of ACD mice were detected by Western Blot.2 acupuncture,yiqi functions of soup of ACD model mice il-17 in the diseased tissue mRNA and Foxp3 mRNA expression effect is not obvious,but the two share can reduce the ACD model il-17 mRNA expression in diseased tissue in mice,increased Foxp3 mRNA expression,this result is inconsistent with PT-PCR detection,the happen reason remains to be further exploration in future research.4.2.2 confirmed that loratine suspension could reduce the expression of il-17 mRNA in the lesion tissues of ACD model mice,and increase the expression of Foxp3 mRNA,acupuncture and beneficial gas disporting decoction and the effect similar to that of loratadine. |
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