复方泽漆冲剂通过抑制NLRP3炎症小体活化改善咪喹莫特诱导银屑病样小鼠的作用及机制研究  

Study on the Effect and Mechanism of Compound Zeqi Granules on Inhibiting theActivation of NLRP3 Inflammasome and Improving Psoriasis-Like Mice Induced by Imiquimod

作　　者：吴敏 刘涛峰[2] 刘小平[2] 曹宇[2] 章纬[2] 汪长中[1,3,4] WU Min;LIU Taofeng;LIU Xiaoping;CAO Yu;ZHANG Wei;WANG Changzhong(School of Integrated Chinese and Western Medicine,Anhui University of Chinese Medicine,Hefei 230012 Anhui,China;The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031 Anhui,China;Institute of Integrated Chinese and Western Medicine,Anhui Academy of Chinese Medicine,Hefei 230012 Anhui,China;Anhui University of Chinese Medicine,Anhui Province Key Laboratory of Traditional Chinese Medicine Compound,Hefei 230012 Anhui,China)

机构地区：[1]安徽中医药大学中西医结合学院,安徽合肥230012 [2]安徽中医药大学第一附属医院,安徽合肥230031 [3]安徽省中医药科学院中西医结合研究所,安徽合肥230012 [4]安徽中医药大学中药复方安徽省重点实验室,安徽合肥230012

出　　处：《中药新药与临床药理》2023年第11期1551-1558,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：安徽中医药大学第一附属医院临床科学研究项目(2020yfyzc18)。

摘　　要：目的基于核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体探讨复方泽漆冲剂(泽漆、大青叶、白花蛇舌草、土茯苓等)对咪喹莫特诱导银屑病样小鼠的作用及机制。方法将56只BALB/c小鼠随机分为正常组、模型组、甲氨蝶呤组(阳性药组)、NLRP3抑制剂组(MCC950组)及复方泽漆冲剂低、中、高剂量组,每组8只。小鼠背部脱毛区涂抹5%咪喹莫特软膏诱导银屑病样小鼠模型,连续7 d。造模同时给药干预,复方泽漆冲剂低、中、高剂量组按照9.75、19.5、29.25 g·kg^(-1)剂量灌胃给药,甲氨蝶呤组按照1 mg·kg^(-1)剂量灌胃给药,各组灌胃体积为10 mL·kg^(-1);正常组和模型组灌胃等体积生理盐水;NLRP3抑制剂组按照25 mg·kg^(-1)剂量腹腔注射MCC950;每天给药1次,连续7 d。采用PASI(Psoriasis area and severity index)评分进行银屑病模型小鼠皮损指数评价;HE染色法观察皮损组织病理变化;免疫组织化学法检测皮损组织NLRP3蛋白表达水平;ELISA法检测小鼠血清白细胞介素1β(IL-1β)、IL-18含量;qRT-PCR及Western Blot法检测皮损组织NLRP3、凋亡相关斑点样蛋白(ASC)、半胱氨酸蛋白酶1(Caspase-1)mRNA及蛋白表达水平。结果与正常组比较,模型组小鼠皮损显著增厚,并出现典型红斑、浸润及鳞屑,PASI评分显著升高(P<0.01);皮肤角化过度伴角化不全,棘层增厚、表皮突下延,真皮浅层淋巴细胞浸润明显,表皮厚度显著增加(P<0.01);血清IL-1β、IL-18水平显著升高(P<0.01);皮损组织NLRP3、ASC、Caspase-1 mRNA及蛋白表达显著上调(P<0.01)。与模型组比较,给药组小鼠背部造模区皮损红斑及鳞屑明显减少,浸润程度明显减轻,PASI评分显著降低(P<0.05,P<0.01),皮肤组织病理损伤程度明显减轻,血清IL-18水平均显著降低(P<0.05,P<0.01),皮损组织NLRP3、ASC、Caspase-1 mRNA及蛋白表达均显著下调(P<0.05,P<0.01);复方泽漆冲剂高剂量组、甲氨蝶呤组及NLRP3抑制剂组小鼠的表Objective To explore the effects and mechanisms of the Compound Zeqi Granules s(Euphoribiae Helioscopiae Herba,Isatidis Folium,Hedyotis Diffusae Herba,Smilacis Glabrae Rhizoma,etc.)on Imiquimod-induced psoriasis-like mice based on nucleotide-binding oligomerisation structural domain-like receptor protein 3(NLRP3)inflammasome.Methods Fifty-six BALB/c mice were randomly divided into normal group,model group,Methotrexate group(positive drug group),NLRP3 inhibitor group(MCC950 group),and Compound Zeqi Granules low-,medium-,and high-dose groups,with 8 mice in each group.The psoriasis-like mouse model was induced by applying 5%Imiquimod ointment to the hairless area on the back of the mice for 7 days.Pharmacological interventions were administered at the same time,with the Compound Zeqi Granules low-,medium-,and high-dose groups being administered by gavage at the doses of 9.75,19.5,and 29.25 gkg^(-1),and the Methotrexate group being administered by gavage at the dose of 1 mg·kg^(-1);and the volume of each group was determined by gavage at the dose of 10 mL·kg^(-1);the normal group and the model group were administered saline in equal volume by gavage;the NLRP3 inhibitor group was administered MCC950 at a dose of 25 mg•kg^(-1)by intraperitoneal injection;the drug was administered once a day for 7 days.The PASI(Psoriasis area and severity index)score was used to evaluate the skin lesion index of psoriasis model mice;HE staining was used to observe the pathological changes of skin lesions;NLRP3 protein expression was detected by immunohistochemistry in skin lesions;serum levels of interleukin 1β(IL-1β)and IL-18 were detected by ELISA;mRNA and protein expressions of NLRP3,apoptosis-associated speck-like protein(ASC),cysteine protease 1(Caspase-1)were detected by qRT-PCR and Western Blot.Results Compared with the normal group,the skin lesions of mice in the model group were significantly thickened with typical erythema,infiltration and scaling,and the PASI scores were significantly elevated(P<0.01);the skin was hyper

关 键 词：复方泽漆冲剂 咪喹莫特 寻常型银屑病 核苷酸结合寡聚化结构域样受体蛋白3炎症小体 炎症因子 小鼠 

分 类 号：R285.5[医药卫生—中药学]