基于Caspase-1/GSDMD蛋白通路探究青石止痒软膏对银屑病小鼠模型的干预机制  

Exploration of the Intervention Mechanism of Qingshi Anti-itch Ointment (青石止痒软膏) on Psoriasis Model Mice Based on Caspase-1/GSDMD Protein Pathway

作　　者：李雅瞳 李元文 邓宇童 任雪雯 王雪皖 于心荟 刘唐云妮 LI Yatong;LI Yuanwen;DENG Yutong;REN Xuewen;WANG Xuewan;YU Xinhui;LIU Tangyunni(Beijing University of Chinese Medicine,Beijing,100029;Dongfang Hospital,Beijing University of Chinese Medicine)

机构地区：[1]北京中医药大学,北京市100029 [2]北京中医药大学东方医院

出　　处：《中医杂志》2025年第2期170-177,共8页Journal of Traditional Chinese Medicine

基　　金：北京市自然科学基金(7232295);北京中医药“十四五”重点专科项目(BJZKBC0015)。

摘　　要：目的 探讨青石止痒软膏治疗银屑病的可能作用机制。方法 将40只小鼠随机分成空白组、模型组、卡泊三醇组和青石组,每组10只。除空白组外,其余各组小鼠均采用咪喹莫特乳膏背部皮肤外用建立银屑病小鼠模型。每日造模6 h后,卡泊三醇组和青石组分别在经造模处理的背部皮肤处给予卡泊三醇软膏0.5 g、青石止痒软膏0.5 g外用治疗,空白组和模型组不干预。隔日观察小鼠皮损变化并进行银屑病面积和严重程度指数(PASI)评分,7日后取背部皮肤组织HE染色观察组织病理学改变,ELISA法测定白细胞介素1β(IL-1β)、白细胞介素18(IL-18)含量,Western Blot法检测含半胱氨酸的天冬氨酸蛋白酶1前体(Pro-Caspase-1)、含半胱氨酸的天冬氨酸蛋白酶1(Caspase-1)、消皮素D(GSDMD)和消皮素D氮端活性结构域多肽(GSDMD-N)蛋白水平,免疫组化法观测GSDMD蛋白水平。结果 与空白组比较,模型组小鼠皮肤变红,出现红斑、白色鳞屑,镜下可见表皮增厚、棘突延长和炎症细胞浸润,第1、3、5、7天皮肤组织PASI评分升高,GSDMD蛋白的IOD、AOD值升高,Caspase-1、Pro-Caspase-1、GSDMD、GSDMD-N蛋白水平及IL-18、IL-1β水平升高(P<0.05或P<0.01)。与模型组比较,青石组和卡泊三醇组皮损较轻;青石组第5、7天及卡泊三醇组第3、5、7天PASI评分降低;两组GSDMD蛋白IOD、AOD值,Caspase-1、GSDMD、GSDMD-N蛋白水平,以及IL-18、IL-1β水平均下降;卡泊三醇组Pro-Caspase-1蛋白水平亦下降(P<0.05或P<0.01)。与卡泊三醇组比较,青石组小鼠皮肤整体稍红,角质层增厚明显,毛细血管扩张现象减轻;第3、7天PASI评分升高,第5天PASI评分降低;Pro-Caspase-1、GSDMD、GSDMD-N蛋白及IL-18、IL-1β水平升高(P<0.05)。结论 青石止痒软膏可有效缓解小鼠银屑病皮炎反应,其作用机制可能与调节Caspase-1/GSDMD蛋白通路,下调IL-18、IL-1β水平,减轻细胞焦亡相关。Objective To explore the possible action mechanism of Qingshi Anti-itch Ointment(青石止痒软膏,QAO)in the treatment of psoriasis.Methods Forty mice were randomly divided into four groups,blank group,model group,calcipotriol group and QAO group,with 10 mice in each group.Except for the blank group,psoriasis was induced by applying imiquimod cream to the dorsal skin.After modeling for 6 hours daily,the calcipotriol group and QAO group were treated with 0.5 g of calcipotriol ointment or 0.5 g of QAO,respectively,applied to the treated dorsal skin.The blank group and the model group received no treatment.The skin lesions were observed,and the psoriasis area and severity index(PASI)score was assessed every other day.After 7 days,Hematoxylin and Eosin(HE)staining was performed on dorsal skin tissue to observe pathological changes.The levels of interleukin 1β(IL-1β)and interleukin 18(IL-18)were determined by enzym-linked immunosorbent assay(ELISA).The protein levels of Caspase-1,Pro-Caspase-1,gasdermin D(GSDMD)and gasdermin-D-N(GSDMD-N)were detected by Western Blot(WB).The protein levels of GSDMD were observed by immunohistochemistry.Results Compared with the blank group,the model group mice showed redness,erythema,and white scales on their skin,with histological observations indicating epidermal thickening,elongated spines,and infiltration of inflammatory cells.The PASI scores of the skin tissue on days 1,3,5,and 7 were elevated;the IOD and AOD values of GSDMD protein increased;the protein levels of Caspase-1,Pro-Caspase-1,GSDMD,GSDMD-N,and IL-1βand IL-18 were significantly elevated(P<0.05 or P<0.01).Compared with the model group,the QAO group and calcipotriol group showed lighter skin lesions;the PASI scores on day 5 and day 7 in the QAO group,and on day 3,5,and 7 in the calcipotriol group,were reduced;the IOD and AOD values of GSDMD protein,and the protein level of Caspase-1,GSDMD,and GSDMD-N,as well as level of IL-18 and IL-1βdecreased in both groups;in the calcipotriol group,Pro-Caspase-1 protein level als

关 键 词：银屑病 青石止痒软膏 含半胱氨酸的天冬氨酸蛋白酶1 消皮素D 

分 类 号：R75[医药卫生—皮肤病学与性病学]