摘要
目的采用病证结合评价方法对比研究3种阳虚型STC大鼠模型建立方法,为中医药防治提供理想的实验动物模型。方法64只SD大鼠,分为空白组和模型组,采用盐酸洛哌丁胺结合0℃冰水复合灌胃法(M-1组)、盐酸洛哌丁胺联合大黄水煎液灌胃法(M-2组)、白醋与0℃冰水交替灌胃+盐酸洛哌丁胺灌胃法(M-3组),各16只,分别建立阳虚型STC大鼠模型,空白组(16只)给予等体积蒸馏水灌胃,造模期间观察大鼠一般情况,每周固定时间测定肛温、饮食量、自主活动量、粪便Bristol评分及含水率,模型建立后检测各组大鼠首粒黑便排出时间、肠道推进率,HE染色法观察大鼠结肠病理改变,PAS染色检测大鼠结肠黏液变化,ELISA法、免疫组化法检测结肠5-HT、VIP、SP含量及蛋白表达。结果造模后各模型组大鼠均出现精神萎靡、倦怠懒动、弓背蜷缩等表现。造模期间各模型组大鼠出现死亡,死亡率依次为M-3组>M-2组>M-1组。与空白组相比,M-1组在造模第3周进食量、自主运动量明显降低(P<0.01),造模第4周肛温、粪便含水率明显降低(P<0.01),造模第5周粪便Bristol评分明显降低(P<0.01);M-2组在造模第3周肛温明显下降(P<0.01),进食量、自主运动量明显降低(P<0.01),第6周粪便Bristol评分、含水率明显降低(P<0.01);M-3组在造模第1周自主运动量明显降低(P<0.01),造模第2周进食量明显降低(P<0.01),造模第4周肛温、粪便Bristol评分、粪便含水率明显下降(P<0.01);HE染色显示各模型组大鼠结肠无明显病理改变;与空白组相比,PAS染色发现各模型组大鼠结肠黏液层厚度明显减少,排序依次为M-3<M-1<M-2,同时各造模组大鼠首粒黑便排出时间增加、肠道推进率显著降低(P<0.01),结肠5-HT、SP含量及蛋白表达显著下降(P<0.01),M-1组结肠VIP含量及蛋白表达显著升高(P<0.01)。结论3种造模方法均能建立阳虚型STC大鼠模型,但以盐酸洛哌丁胺联合0℃冰水灌胃法建立的大鼠模型具有死亡率低、安全性较好、阳虚证候表现稳定,大便性状改变明显,肠道动力障碍病理变化显著的优点,是阳虚型STC发病机制及相关中医药药物作用实验研究的理想模型。
Objective To compare three kinds of yang-deficiency STC rat models by a combination of disease and syndrome evaluation to provide an ideal experimental animal model for TCM prevention and treatment of this disease.Methods Sixty-four SD rats were divided into blank and model groups.The rats were treated with loperamide hydrochloride combined with 0℃ice water by gavage(M-1 group),loperamide hydrochloride combined with dahuang decoction(M-2 group),and white vinegar and 0℃ice water by gavage alternately+loperamide hydrochloride by gavage(M-3 group)(16 rats in each group)to establish the three yang-deficiency slow transit constipation rat models.The blank group(n=16)was administered the same volume of distilled water by gavage.Model groups were established by the corresponding method.The general condition of rats was observed during modeling.Rectal temperature,food intake,voluntary activity,stool Bristol score,and water content were measured at fixed times every week.After establishing the model,the discharge time of the first black stool and the intestinal propulsion rate were measured.Rat colon pathological changes were observed by HE staining.Colon mucous changes were assessed by PAS staining.Colon 5-HT,VIP,and SP were analyzed by ELISA and IHC.Results Blank group rats before and after building without exception.After modeling,rats in each model group showed listlessness,lethargy,and crouching.Some rats died during modeling.Mortality was in the order of M-3 group>M-2 group>M-1 group.Compared with the blank group,the anal temperature,food intake,and voluntary activity of each model group showed a downward trend during modeling.The model groups were compared with the blank group as follows:M-1 group:building 3 weeks of food intake,autonomous activity was obviously decreased(P<0.01),building 4 weeks of rectal temperature and fecal water content were significantly decreased(P<0.01),building 5 weeks of Bristol stool scale was significantly decreased(P<0.01).M-2 Group:building 3 weeks of rectal temperature,food intake,and autonomous activity were significantly decreased(P<0.01),building 6 weeks of the Bristol stool scale and fecal water content were significantly decreased(P<0.01).M-3 Group:building 1 week of autonomous activity was decreased significantly(P<0.01),building 2 weeks of food intake significantly decreased(P<0.01),building 4 weeks of rectal temperature,Bristol stool scale,and fecal water content were significantly decreased(P<0.01).HE staining showed no obvious pathological changes in the colon of rats in each group after modeling.PAS staining showed that the mucus layer thickness in the colon of rats in each model group was significantly reduced(M-3<M-1<M-2).Compared with the blank group,the first black stool excretion time in the model groups was increased,and the intestinal propulsion rate,and 5-HT and SP in the colon were significantly decreased(P<0.01).VIP in the colon of M-1 group was significantly increased(P<0.01).Conclusions The STC model of yang-deficiency can be established by the three modeling method.The rat model established by intragastric administration of loperamide hydrochloride combined with 0℃ ice water has the advantages of low mortality,good safety,stable yang-deficiency syndrome,obvious changes in stool characteristics,and significant pathological changes in intestinal motility.It is an ideal model to study the STC pathogenesis of yang-deficiency and the effect of traditional Chinese medicines.
作者
张远哲
赵罗娜
简胜男
李中正
黎豫川
ZHANG Yuanzhe;ZHAO Luona;JIAN Shengnan;LI Zhongzheng;LI Yuchuan(Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;Chengdu University of TraditionalChinese Medicine,Chengdu 620005;Jishou University,Xiangxi Tujia and Miao Autonomous Prefecture 416000;the Second Affiliated Hospital of Guizhou University of Chinese Medicine,Guiyang 550025)
出处
《中国实验动物学报》
CAS
CSCD
北大核心
2023年第5期618-628,共11页
Acta Laboratorium Animalis Scientia Sinica
基金
贵州省省级科技计划项目资助(黔科合基础[2020]1Y362,黔科合基础-ZK[2022]一般510)
贵州省中医药、民族医药科学技术研究专项课题项目(QZYY-2021-016)
贵州省教育厅青年科技人才成长项目(黔教合KY字[2022]261号)。
关键词
阳虚型慢传输型便秘
大鼠模型
病证结合
yang-deficiency slow transit constipation
rat model
disease combined traditional Chinese medicine evidence
