| Objective:In this study,we induced urticaria rats model by the injection methods of chicken egg innocence protein,dipperazine vaccine and Evans blue,observed the effects of angelica decoction on autophagy and related proteins in urticaria rats,and revealed the mechanism of angelica decoction in the treatment of urticaria.Methods:50 SD rats,half male and half female.Ten rats randomly selected were injected with normal saline solution of egg white protein in all limbs and feet,and intraperitoneally injected with white peroxazine vaccine,once every other day for a total of 3 times.They were killed after 10 d of the last sensitization,and blood was collected from abdominal aorta to prepare serum.40 rats were randomly divided into blank group,model group,western medicine control group and angelica decoction group,with 10 rats in each group.Normal saline was given to the blank group and the model group,and loratadine solution and angelica decoction were given to the western medicine control group and angelica decoction group respectively.Each group received 5consecutive days of drug intervention,1 time per day.On the 5th day of intervention,rats were shaved on the back,and normal saline was injected intradermal at the shearing site in the blank group,and serum diluent was injected intradermal at the shearing site in the other 4 groups.Intravenous injection of Evans blue back 1 hour add chicken egg white protein physiological saline solution,put to death animals,take skin tissue for HE staining to observe the histopathological changes,transmission electron microscope autophagy morphological,immunohistochemical staining method to detect LC3 B,beclin-1,p62 protein expression,using the Image Pro Plus6.0 protein IOD semi-quantitative score Image software,statistical analysis by SPSS25.0.Results:Histopathology:The epidermal structure of the blank group was intact,and no obvious pathological changes were observed.In the model group,significant dermal edema was observed,collagen fiber staining was weakened,interfiber space was widened,light transmittance was enhanced,vascular dilation and inflammatory cell infiltration were observed,and histopathological changes like urticaria were observed,which were alleviated in all treatment groups.Electron microscopy(sem):No obvious autophagosome formation was observed in the blank group.In the model group,the number of autophagosomes increased,and abnormal ultrastructures of epithelial cells,such as nuclear fragmentation,mitochondrial swelling and damage accompanied by vacuolar degeneration and endoplasmic reticulum expansion were observed.The number of autophagosomes and autophagic lysosomes in the angelica decoction group was significantly higher than that in the model group,and angelica decoction could improve the abnormal ultrastructure of epithelial cells in skin lesions.Immunohistochemical staining:Compared with the blank group,LC3 B expression in the model group was significantly increased(P < 0.01).Compared with the model group,LC3 B protein expression level in skin tissues of western medicine control group and angelica decoction group was increased(P<0.01).Compared with western medicine control group,angelica decoction subgroup could up-regulate the expression of LC3 B protein(P<0.01).Compared with the blank group,beclin-1 expression in the rats model group was significantly increased(P< 0.01).Compared with the rats model group,the expression of beclin-1 in the skin lesions of the danggui decoction group was further increased(P< 0.05).Compared with the blank control group,p62 expression in the model group was significantly reduced(P < 0.05).Compared with the model group,the expression level of p62 in the skin lesions of the danggui decoction group was decreased(P <0.05).Compared with the western medicine control group,angelica decoction can down-regulate the expression level of p62 in rats skin lesions(P < 0.05),suggesting that after treatment with danggui decoction,the expression levels of autophagy markers LC3 B and beclin-1 in urticaria rats model were increased,while the expression level of p62 protein was decreased.Conclusion:1.Chicken egg white protein complex modeling method successfully prepared urticaria rats model,danggui decoction can significantly improve the general situation and histomorphological changes of urticaria rats model;2.The number of autophagy in skin epithelial cells of urticaria model rats increased,and angelica decoction could further promote the activation and formation of autophagy,improve the level of autophagy and improve the abnormal ultrastructure of epithelial cells in skin lesions;3.Angelica decoction can up-regulate the expression levels of LC3 B and beclin1 protein,and down-regulate the expression levels of p62,which may be one of the mechanisms of angelica decoction in the treatment of urticaria.Based on the above conclusions,angelica decoction may promote the level of autophagy in the skin tissues of urticaria rats,so as to achieve the goal of curing urticaria. |
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