Effect Of Yangxuehuoxuejiedu Recipe On The Expression Of PI3K/AKT/β-catenin Pathway In Skin Lesions Of Psoriasis-like Model Mic

Objective:To observe the effect of Yangxuehuoxuejiedu Decoction on psoriasis through PI3K/AKT/β-catenin pathway and angiogenesis and apoptosis involved protein in the expression of Survivin and VEGF.At the same time,we observe the autophagy expression in psoriatic model mice and the intervention of Yangxuehuoxuejiedu Decoction on psoriatic model mice which could investigate the possible mechanism of Yangxuehuoxuejiedu Decoction in the treatment of psoriasis with angiogenesis,apoptosis as well as autophagy.Methods:Twenty-two BALB/c mice were randomly divided into blank control group,model group,Yangxuehuoxuejiedu Decoction group and Cyclosporin group.Except the blank control group,the other groups were continuous coated with 5%Irniquimod Cream for 5 days to establish psoriasis model.The blank control group and the model group were given with distilled water once a day respectively.And the Cyclosporin group were given with Cyclosporin capsules(3 times per day,80mg/kg),and Yangxuehuoxuejiedu Decoction group was given 10 ml/kg medicine twice a day.On the seventh day,we observed the changes in the lesions of the mouse’s back.The psoriasis area and severity index(PASI)were used for scoring of the lesions,HE staining was used to observe the morphological changes of the lesions,and the thickness of the epidermis was measured under the light microscope,and the number of microvessels in the skin lesions was observed under light microscope.Western blot was used to detect the proteins related to PI3K/AKT/β-catenin pathway,as well as VEGF and Survivin proteins,to compare the differences in protein contents between different groups,and immunofluorescence was used to determine the expression positions of VEGF and Survivin in the skin and the entrance of β-catenin into nuclear.Meanwhile,Western blot was used to detect the expressions of proteins LC3Ⅱ/Ⅰ and p62 which were related to autophagy.Results:1.Compared with the model group,the PASI score of Yangxuehuoxuejiedu Decoction group and Cyclosporin group showed significant differences(P<0.05),and the thickness of the epidermis and the number of microvessels showed statistically significant differences(P<0.01).There was no statistically significant difference between Yangxuehuoxuejiedu Decoction group and Cyclosporin group.2.Compared with the model group,the levels of PI3K/Akt/β-catenin signaling pathway related proteins,VEGF and Survivin were significantly down from Yangxuehuoxuejiedu Decoction group and Cyclosporin group,which showed statistically significant differences.The down-regulation effect of β-catenin protein in Cyclosporin group was better than that in Yangxuehuoxuejiedu Decoction group,with statistical difference(P<0.05).There was no statistical difference in the regulation of other proteins between Yangxuehuoxuejiedu Decoction group and Cyclosporin group.3.The expression of LC3Ⅱ/Ⅰ protein in psoriasis mouse lesions was lower than that in the control group,while the expression of p62 protein was higher than that in the control group(P<0.05).In mice treated with Yangxuehuoxuejiedu Decoction and Cyclospori,the expression of LC3 Ⅱ/Ⅰ protein in the skin tissue on the smear was significantly increased,while the expression of p62 protein was significantly decreased(P<0.05).Conclusion:1.The expression levels of PI3K/Akt/β-catenin signal pathway related proteins and VEGF and Survivin in lesions of psoriasis mouse model group were higher than blank group,indicating that the high expression of this pathway protein,VEGF and Survivin can lead to microvascular abnormalities in psoriasis mouse model,which confirmed that the pathogenesis of psoriasis is related to abnormal microangiogenesis and apoptosis induced by PI3K/Akt/catenin signal pathway.2.Yangxuehuoxuejiedu Decoction may inhibit AKT,GSK3-β and β-catenin expression,activation of PI3K/AKT β-catenin signaling pathways play a role,and then cut the expression of VEGF and Survivin protein,downstream products applied to model on the target lesion of microvascular endothelial cells in mice,inhibit angiogenesis,apoptosis of blood vessels,and effective treatment for psoriasis.Meanwhile,we used immunofluorescence method to determine the expression of VEGF and Survivin and the nuclear entry of β-catenin.The Yangxuehuoxuejiedu Decoction and Cyclosporin capsules could down-regulate the expression of VEGF and Survivin in skin vascular endothelial cells of psoriasis model mice,and reduce the expression and entry of β-catenin3.The autophagy level in the skin tissue of the psoriasis model mice was significantly reduced,and the administration of Yangxuehuoxuejiedu Decoction could significantly inhibit the autophagy level in the skin tissue of the psoriasis model mice.