基于腺苷酸活化蛋白激酶—结节性硬化复合物1/2信号通路研究加味蜈蚣败毒饮在复发型银屑病中的作用机制OA

Objective To explore the mechanism by which the Modified Wugong Baidu Drink inhibits psoriasis recurrence by affecting the upstream autophagy signaling pathway AMPK-TSC1/2.Methods 30 mice were used to establish recurrent psoriasis model mice by a twice induction with imiquimod method(except for the blank control group).Specifically,imiquimod cream(62.5 mg/d)was topically applied for 8 consecutive days,followed by a 30-day natural recovery period,and then a secondary application for 8 days at the site where the original skin lesions had subsided.The model mice were randomly divided into the model control group,western medicine control group,and low,medium and high dose groups of traditional Chinese medicine,with 5 mice in each group.The mice in each group received drug interventions via gavage.The low-,medium-,and high-dose traditional Chinese medicine groups were administered the Modified Wugong Baidu Drink at concentrations of 0.1 g/mL,0.2 g/mL,and 0.4 g/mL,respectively.The western medicine control group received methotrexate suspension(0.003 mg/mL)for the first 4 days,followed by normal saline for the next 4 days.The model control group was given an equal volume of normal saline,while the blank control group remained untreated.The gavage dose was 1 mL per 20 g of body weight,administered twice daily for 8 consecutive days(days 47～54).After the gavage period(day 54),samples were collected,specifically the typical skin lesion tissue from the back of the mice,for subsequent analysis.The MPASI scoring method was used to evaluate the skin lesion status of mouse;Using TUNEL method to stain apoptotic cells in mouse skin lesions;Methods of western blotting and immunohistochemical staining were used to detect the content,expression and distribution of AMPKα,TSC1 and TSC2 proteins in mouse skin lesions.Results(1)When imiquimod was induced for the second time,the appearance time of psoriasis like lesions in mice was earlier than that of the first induction(2d＜4d),and the macroscopic appearance of the skin lesions becomes more severe when the modeling is completed.(2)Before drug intervention in the recurrence model,the MPASI scores were all above 24 points,All dose groups of traditional Chinese medicine can sig-nificantly improved the MPASI score of the recurrent psoriasis model,the curative effect of the high dose group was particularly significant(P＜0.05).(3)After drug intervention,there was a significant increase in apoptotic cells located in the stratum corneum and granular layer of recurrent skin lesions,with apoptosis being the dominant factor,thereby improving the condition of thickened skin lesions.(4)All dose groups of traditional Chinese medicine can regulate the AMPK-TSC1/2 signaling pathway,this regulatory effect includes inhibiting AMPKα proteins(P＜0.05)and TSC1 proteins(P＜0.05),promoting TSC2 protein(P＜0.05),and intervening the content,expression,and distribution of these proteins in the skin lesion tissue of the recurrence model,thereby affecting the level of related mechanism pathways for psoriasis recurrence,among them,the high dose group of traditional Chinese medicine has the most significant effect(P＜0.05),and the results of the content and expression of the above mentioned proteins by WB method were consistent with those by immunohistochemical method.Conclusion Modified Wugong Baidu Drink may regulate the key signaling pathway AMPK-TSC1/2 upstream of autophagy,affecting the energy supply of TRM to autophagy response,thereby inhibiting the reactivation of cells in skin lesions and reducing the recurrence of psoriasis.