摘要
目的建立湿证小鼠模型(单模)并观察、评价其湿证特征,在湿证小鼠模型基础上合并构建缺血性中风模型(双模),并观察湿邪对中风结局的影响。方法将C57BL/6雄性小鼠随机分为湿证组(包括湿证假手术组、湿证缺血性中风组,每组10只)、非湿证组(包括非湿证假手术组、非湿证缺血性中风组,每组10只)。湿证组给予高脂饲料饲养,非湿证组给予普通饲料饲养,连续饲养12周。复制湿证小鼠模型后,采用大脑中动脉短暂阻塞/再灌注(tMCAO/R)术复制缺血性中风小鼠模型。湿证小鼠模型评价指标:体质量、形态、神态、活动状态、体表特征等一般形态,以及主动脉组织病理学观察(油红O染色、Masson三色染色)、肝脏组织病理学观察(HE染色、油红O染色)、舌组织电镜观察(扫描电镜、透射电镜)、血脂[总胆固醇(TC)、甘油三酯(TG)]水平、肝脏系数。缺血性中风小鼠模型评价指标:神经功能评分、脑梗死体积比。结果与非湿证组比较,湿证组小鼠体型肥胖,毛发色泽差,毛发稀疏,倦怠懒动,精神萎糜,纳呆嗜睡,体质量显著升高(P<0.001);主动脉管腔变窄、内膜明显增厚,脂质斑块沉积增多,内含泡沫细胞;肝细胞出现大量红色脂滴,脂质浸润明显,弥漫性脂肪变性;舌组织黏膜层角化增加,角质层较厚,脂褐素、舌体表面细菌较多;血清TG、TC水平显著升高(P<0.01);肝脏系数显著降低(P<0.001)。与非湿证假手术组比较,湿证缺血性中风组小鼠的神经功能评分及脑梗死体积比显著升高(P<0.001)。结论高脂喂养12周联合tMCAO/R术成功建立湿证缺血性中风小鼠模型,且湿证中风模型小鼠在神经功能评分及脑梗死体积方面均较非湿证中风组更加严重。
Objective To establish an animal model of dampness-syndrome in mice(single model)and evaluate its characteristics of dampness-syndrome.The above-mentioned mice with dampness syndrome were used to construct mice model of ischemic stroke(double model)and observe the effect of dampness-pathogenic on the outcome of stroke.Methods Healthy C57BL/6J male mice were randomly divided into dampness-syndrome(including sham-surgery group and ischemic stroke group,with 10 mice in each group)and non dampnesssyndrome groups(including sham-surgery group and ischemic stroke group,with 10 mice in each group).The dampness-syndrome group was fed with high-fat diet and the non dampness-syndrome group was fed with normal diet for 12 weeks.After the mice model of dampness-syndrome was successfully established,transient middle cerebral artery occlusion/reperfusion(tMCAO/R)surgery was used to replicate an ischemic stroke mice model.Evaluation indicators for dampness-syndrome mice model:the general status including body weight,morphology,posture,activity status,and physical characteristics,the histopathological observation of the aorta(oil red O staining,Masson-trichrome staining)and liver(HE staining,oil red O staining),electron microscopic observation of the tongue tissue(scanning electron microscopy,electron microscopy),blood lipid levels[total cholesterol(TC),triglycerides(TG)]and liver coefficient.Evaluation indicators for ischemic stroke mice model:neurological function score and the cerebral infarction volume ratio.Results Compared with the non dampnesssyndrome group,the mice in the dampness-syndrome group showed an increased in body weight,poor hair color,sparse hair,fatigue and laziness,mental atrophy,anorexia and lethargy.It was observed that the aortic lumen was narrowed,the intima was significantly thickened,lipid plaque deposition was increased,and foam cells were visible.A large amount of red lipid droplets appeared in liver cells.There were obvious lipid infiltration and diffuse steatosis.Increased keratosis of the mucosal layer of tongue tissue,the thicker stratum corneum,lipofuscin,and bacteria on the tongue surface were found.Serum TG and TC levels significantly increased(P<0.01),and the liver coefficient significantly decreased(P<0.001).Compared with non dampness-syndrome group(sham-surgery group),neurological function score and the cerebral infarction volume ratio in dampness-syndrome ischemic stroke group obviously increased(P<0.001).Conclusion High-fat feeding for 12 weeks combined with tMCAO/R modeling can successfully establish a mice model with dampness-syndrome ischemic stroke,and the neurological function score and cerebral infarction volume in the dampness-syndrome ischemic stroke group was more severe than that in the non dampness-syndrome ischemic stroke group.
作者
李昆宏
武爽
杨嘉葳
王宇
王亚琼
邓敏贞
黄燕
孙景波
黎创
李岩
程骁
LI Kunhong;WU Shuang;YANG Jiawei;WANG Yu;WANG Yaqiong;DENG Minzhen;HUANG Yan;SUN Jingbo;LI Chuang;LI Yan;CHENG Xiao(Guangdong Provincial Hospital of Chinese Medicine/The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120 Guangdong,China;The Second Clinical Medical College of Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China;Zhongshan Medical College,Sun Yat-sen University,Guangzhou 528406 Guangdong,China;State Key Laboratory of Traditional Chinese Medicine Syndrome,Guangzhou 510120 Guangdong,China;State Key Laboratory of Dampness Syndrome of Chinese Medicine,Guangzhou 510120 Guangdong,China;Key Research Laboratory of Emergency Research in Traditional Chinese Medicine,Guangzhou 510120 Guangdong,China;Guangxi International Zhuang Medical Hospital,Nanning 530201 Guangxi,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2024年第10期1492-1497,共6页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金项目(8216150148)
广州市科技计划项目(202102010268)
省部共建中医湿证国家重点实验室专项(SZ2021ZZ14,SZ2023ZZ04)
广东省中医急症重点实验室项目(2023B1212060062)。
关键词
湿证
缺血性中风
动物模型
评价指标
小鼠
Dampness syndrome
ischemic stroke
animal model
evaluation index
mice
