痤疮脾胃湿热病证结合小鼠模型的建立与评价  

Establishment and evaluation of a mouse model combining acne and damp-heat syndrome of spleen and stomach

作　　者：任星 周娜 王羽侬 杨静[3] 李俐霜 马语蔓 贺改英 马淑骅[1] 孙娅楠[1] 王毅[1] REN Xing;ZHOU Na;WANG Yunong;YANG Jing;LI Lishuang;MA Yuman;HE Gaiying;MA Shuhua;SUN Yanan;WANG Yi(Experimental Research Center,China Academy of Chinese Medical Sciences,Beijing 100700,China;Beijing University of Chinese Medicine Third Affliated Hospital,Beijing 100029,China;Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China)

机构地区：[1]中国中医科学院医学实验中心,北京100700 [2]北京中医药大学第三附属医院,北京100029 [3]中国中医科学院西苑医院,北京100091

出　　处：《中华中医药杂志》2024年第9期4959-4964,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：中国中医科学院科技创新工程重大攻关项目(No.CI2021A00601)。

摘　　要：目的:结合疾病病因造模方法与脾胃湿热证候造模方法建立以临床表现为导向的痤疮脾胃湿热病证结合小鼠模型。方法:采用皮内注射痤疮丙酸杆菌(C.acnes)建立痤疮病因模型,结合高脂高糖喂养(内因湿热法)构建脾胃湿热病证结合模型,通过清热利湿中药方剂进行模型反证。比较各组小鼠体质量、行为活动度、肛温、大小便评价脾胃湿热程度;采用皮肤镜、激光散斑血流成像仪活体观测小鼠背部痤疮区域的变化;HE染色观察皮肤和胃黏膜组织的炎症改变,油红O染色观测皮脂分泌情况;免疫组化法观察小鼠皮肤中IGF-1R、PPAR-γ、SREBP-1、AR的表达水平。结果:与空白组和C. acnes组比较,C. acnes+脾胃湿热组小鼠行为活动度减少(P<0.05),反应迟钝,体质量下降,大小便色黄质软,胃黏膜固有层与黏膜下层有炎症细胞浸润;痤疮皮损更严重,痤疮部位的局部血流量显著增加(P<0.01,P<0.05),相关炎症因子表达上调(P<0.01,P<0.05);除此之外,小鼠皮脂腺增大(P<0.01),皮脂分泌增多(P<0.01)。经中药干预后,上述改变均有缓解。与空白组及C. acnes组比较,C. acnes+脾胃湿热组小鼠表皮中IGF-1R表达增多(P<0.01),皮脂腺中的PPAR-γ、SREBP-1、AR阳性表达明显增多(P<0.01),经中药干预后小鼠皮脂腺中的PPAR-γ、SREBP-1、AR阳性表达减少(P<0.01)。结论:高脂高糖饮食喂养小鼠模拟人体脾胃湿热,结合皮内注射C. acnes模拟痤疮炎症发病过程,综合形成了痤疮脾胃湿热小鼠模型,可作为可靠的动物模型,推动痤疮病理机制与中医药疗效评价研究的进展。Objective:To establish a mouse model combining acne and damp-heat syndrome of spleen and stomach,guided by clinical manifestations,through the integration of pathogenesis and pathophysiology modeling methods with modeling methods for spleen and stomach damp-heat syndrome.Methods:The model combining acne and damp-heat syndrome was created by combining intradermal injection of Cutibacterium acnes(C.acnes).with a high-fat and high-sugar diet(internal dampheat method).The model was further verified by the administration of heat-clearing and dampness-resolving Chinese herbal formulations.Various parameters,including body weight,activity level,rectal temperature,and bowel habits,were compared to evaluate the degree of spleen and stomach damp-heat.Skin microscopy and Laser Speckle Contrast Imaging were used to observe changes in the acne area on the backs of mice.HE staining was employed to observe infiammatory changes in the skin and gastric mucosal tissues,and oil red O staining was used to observe sebum secretion.Immunohistochemistry was utilized to observe the expression levels of IGF-1R,PPAR-,SREBP-1,and AR in mice.Results:Compared with the blank group and the C.acnes group,mice in the spleen and stomach damp-heat model group exhibited reduced activity(P<0.05),delayed response,weight loss,yellowish and soft stools,and infiltration of infiammatory cells in the intrinsic and submucosal layers of the gastric mucosa.The acne lesions were more severe,and the local blood flow in the acne area was more pronounced(P<0.01,P<0.05),with upregulation of relevant inflammatory factors(P<0.01,P<0.05).In addition,mice in the spleen and stomach damp-heat model group had enlarged sebaceous glands(P<0.01)and increased sebum secretion(P<0.01).After intervention with traditional Chinese medicine,these changes were alleviated.Compared with the blank group and the C.acnes group,the expression of IGF-IR in the epidermis of mice in the spleen and stomach damp-heat model group increased(P<0.01),and the positive expression of PPAR-,SREBP-1,a

关 键 词：痤疮 脾胃湿热证 病证结合 模型 炎症 皮脂 痤疮丙酸杆菌 

分 类 号：R275.9[医药卫生—中西医结合] R-332[医药卫生—中医皮科]