脾虚湿阻型银屑病病证结合小鼠模型的系统评价研究  

A Systematic Evaluation Study of a Mouse Model of Psoriasis with Pattern of Spleen Deficiency and Dampness Obstruction

作　　者：刘凡露 苏浩杰 周攀宇 张雅婷 汪晴 孙玥 岳洪宇 吴晶晶[1] 危建安[1] 韩凌[1,2,4,5] LIU Fanlu;SU Haojie;ZHOU Panyu;ZHANG Yating;WANG Qing;SUN Yue;YUE Hongyu;WU Jingjing;WEI Jianan;HAN Ling(The Second Clinical Medical School of Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China;State Key Laboratory of Dampness Syndrome of Chinese Medicine,the Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120 Guangdong,China;Dongguan Hospital of Guangzhou University of Chinese Medicine,Dongguan Hospital of Traditional Chinese Medicine,Dongguan 523127 Guangdong,China;Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome,Guangzhou 510120 Guangdong,China;Guangdong-Hong Kong-Macao Joint Lab on Chinese Medicine and Immune Disease Research,Guangzhou 510120 Guangdong,China)

机构地区：[1]广州中医药大学第二临床医学院,广东广州510006 [2]省部共建中医湿证国家重点实验室(广州中医药大学第二附属医院),广东广州510120 [3]广州中医药大学东莞医院(东莞市中医院),广东东莞523127 [4]广东省中医证候临床研究重点实验室,广东广州510120 [5]粤港澳中医药与免疫疾病研究联合实验室,广东广州510120

出　　处：《中药新药与临床药理》2024年第10期1470-1482,共13页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(82374313);广东省科技计划项目(2020B1111100006,2023B1212060063);广州市科技计划项目(202201020332);省部共建中医湿证国家重点实验室重点项目(SZ2021ZZ29);广东省中医院拔尖人才科研专项(BJ2022KY05)。

摘　　要：目的构建脾虚湿阻型银屑病小鼠模型,并从多维度、多方向评价该模型,以期为中医药治疗脾虚湿阻型银屑提供研究支持。方法采用高脂饮食喂养建立脾虚湿阻证小鼠模型,外涂咪喹莫特乳膏建立银屑病小鼠模型,并二者结合构建病证结合小鼠模型。比较体质量、进食量和饮水量,评价脾虚湿阻证候指征。比较皮损面积、PASI评分、经皮水分丢失值、HE染色下皮肤病理学改变评价银屑病严重程度。流式细胞术检测各类细胞表达情况,评价炎症程度。观察脂肪指数、肝脏HE染色下肝脏病理学变化、RT-q PCR法检测肝脏和附睾脂肪的相关因子m RNA表达水平、Western Blot法检测皮肤ABCA1蛋白表达水平,评价脂代谢紊乱情况。结果与寻常型银屑病组小鼠比较,脾虚湿阻型银屑病组小鼠体质量升高(P<0.001),进食量下降(P<0.005),出现毛发油腻,精神萎靡等脾虚湿阻证候指征;经皮水分丢失值升高(P<0.001),PASI评分增加(P<0.001),皮肤HE染色病理结果提示表皮棘层肥厚、杵状增生等银屑病样表现;CD11^(bhigh)Ly6G+中性粒细胞、CD11b^(in)Ly6C^(high)单核细胞、CD11binCD11chigh经典树突细胞、F4/80-CD11c+树突状细胞表达上升(P<0.001);肝脏HE染色病理结果提示细胞空泡样变性,且皮下白色脂肪指数和附睾脂肪指数上升(P<0.005);肝脏FABP4、CD36 m RNA水平升高(P<0.005,P<0.001);附睾脂肪ABCA1和PPARγm RNA水平下降(P<0.05,P<0.01),皮肤ABCA1蛋白水平升高(P>0.05)。结论脾虚湿阻型银屑病小鼠模型可作为可靠的病证结合动物模型,为进一步探讨中医药治疗脾虚湿阻型银屑病提供参考。Objective To construct a mouse model of psoriasis with spleen deficiency and dampness obstruction pattern and evaluate the model from multiple dimensions and directions,expects to provide research support for the study of traditional Chinese medicine(TCM)treatment of psoriasis with spleen deficiency and dampness obstruction pattern.Methods A mouse model of spleen deficiency and dampness obstruction pattern was established by feeding a high-fat diet,a mouse model of psoriasis vulgaris was established by externally applying imiquimod ointment,and a mouse model of psoriasis with spleen deficiency and dampness obstruction pattern was constructed by combining the above two models.Indications of spleen deficiency and dampness obstruction pattern were evaluated by comparing the body mass,food intake and water intake of mice in each group.The severity of psoriasis in mice was evaluated by comparing the area of skin lesions,PASI score,the value of transdermal water loss(TEWL),and histopathological morphological changes of skin under HE staining in each group.Flow cytometry was used to detect the expression in various cell types to evaluate the degree of inflammatory response of psoriasis in mice.Observation of adiposity index,changes in the histopathological morphology of liver tissue under HE staining,changes in the mRNA expression levels of related factors in liver tissue and adipose tissue of epididymis of mice detected by RT-qPCR,and changes of ABCA1 protein expression level of skin detected by Western Blot were used to evaluate the lipid metabolism disorders in mice.Results Compared with the mice in the psoriasis vulgaris model group,the mice in the model of psoriasis with pattern of spleen deficiency and dampness obstruction had significantly higher body mass(P<0.001),significantly lower food intake(P<0.005),and the symptoms of pattern of spleen deficiency and dampness obstruction such as greasy fur,mental fatigue,etc.appeared.The TWEL were significantly increased(P<0.001),and the PASI scores also significantly incr

关 键 词：银屑病 脾虚湿阻证 病证结合 动物模型 小鼠 

分 类 号：R285.5[医药卫生—中药学]