积热饲料喂食联合洛哌丁胺灌胃制作胃肠积热证大鼠模型的优化与验证

Optimization and validation of rat models with gastrointestinal heat accumulation syndrome constructed by high protein and calorie diet and loperamide

目的以积热饲料喂食联合洛哌丁胺灌胃制作胃肠积热证大鼠模型并加以优化和验证,为中医药经典理论研究、药效学评价等提供实验基础。方法选择3周龄乳雌性SD大鼠80只,以积热饲料喂食联合洛哌丁胺灌胃为胃肠积热证大鼠建模方法;取其中60只采用三因素三水平正交设计的方法,以积热饲料喂食时间(4、8、12周)、洛哌丁胺给药剂量(3、6、12 mg/kg)、洛哌丁胺给药时间(5、10、15 d)为主要因素,以舌色、体温、粪便、尿液、肠鸣音、冷热趋向性的宏观表征作为考察指标,优化胃肠积热证大鼠模型的实验条件;将剩余20只分为两组各10只,模型组采用优化后的实验条件制作胃肠积热证大鼠模型,对照组常规饲养、生理盐水灌胃,对比两组宏观表征指标,以验证造模稳定性。结果正交设计实验结果表明,以积热饲料喂食联合洛哌丁胺灌胃制作胃肠积热证大鼠模型积热饲料喂食时间为8周,洛哌丁胺给药剂量为12 mg/kg,洛哌丁胺给药时间为10 d;与对照组比较,模型组大鼠舌色中红色、绿色、蓝色像素值降低,尿液中蓝色像素值降低,粪便性状评分升高,肠鸣音次数减少,在低温区域停留时间延长,直肠温度升高(P均<0.05),符合胃肠积热证诊断标准。结论以积热饲料喂食联合洛哌丁胺灌胃制作胃肠积热证大鼠模型的最佳实验条件为积热饲料喂食时间8周、洛哌丁胺给药剂量12 mg/kg、洛哌丁胺给药10 d,以此所建胃肠积热证大鼠模型具有较好的稳定性。

Objective To provide experimental data and theoretical basis for classical theoretical studies of traditional Chinese medicine and pharmacodynamic evaluation by constructing the rat models with gastrointestinal heat accumulation syndrome.Methods Eighty 3-week-old female preweaned Sprague Dawley(SD)rats were selected to establish the rat models of gastrointestinal heat accumulation syndrome by loperamide,high protein and calorie diet.Sixty rats were randomly divided into ten groups:group Control and groups 1-9.An experiment of orthogonal design[L9(34)]was used for optimizing the three main factors,the duration of high protein and calorie diet(4,8,and 12 w),dosage of loperamide(3,6,and 12 mg/kg),and the duration of loperamide(5,10,and 15 d).The characteristics of tongue color,body temperature,feces,urine,hot and cold tendencies,and bowel sounds were used as indicators of investigation to optimize the experimental conditions.The remaining 20 rats were divided into two groups:model group and control group.In the model group,we adopted the optimized experimental conditions to make the rat models of gastrointestinal heat accumulation syndrome,while the rats in the control group were routinely fed with normal food and saline.Then indicators were observed to verify the stability.Results The best conditions were feeding with high protein and calorie diet for 8 w and 12 mg/kg loperamide for 10 d.The rat model of gastrointestinal heat accumulation syndrome is successfully established.Compared with the control group,the red,green,and blue pixel values of the tongue color of the model group decreased,the blue pixel value of the urine descended,the stool trait score increased,the number of bowel sounds diminished,the time in the hypothermic area increased and the rectal temperature increased(all P<0.05).These results met the diagnostic criteria for gastrointestinal heat accumulation syndrome.Conclusion The best conditions to build a rat model of gastrointestinal heat accumulation syndrome are feeding with high protein and calorie diet for 8 w and 12 mg/kg loperamide for 10 d,and the established gastrointestinal heat accumulation syndrome rat models based on the above conditions have a good stability.