高脂致石饲料联合21天“CUMS+孤养”建立胆石症肝郁证动物模型研究

Study on the Establishment of Animal Model of Cholelithiasis with Liver Stagnation Syndrome by High-Fat Stone-Causing Feed Combined with 21-Day “CUMS+Orphan Rearing”

目的:采用高脂致石饲料联合21天“慢性不可预知温和刺激(CUMS)+孤养”模式,制备胆石症肝郁证动物模型。方法:33只雄性ICR小鼠随机分为对照组12只,致石组21只。不同饮食喂养16周后,每组取3只查看胆囊内结石形成情况、胆汁浑浊度及透光性;酶联免疫吸附测定法(ELISA)检测胆囊胆汁总胆固醇(TC)、总胆汁酸(TBA)及磷脂(PL)含量,判断是否建立胆石症病模型。模型成功后,致石组随机分为模型组(CS组)、模型肝郁组(CS肝郁组)。CS肝郁组采用21天“CUMS+孤养”模式建立肝郁模型。造模期间动态观察小鼠的精神状况、摄食、行为活动,及时记录小鼠体质量、强迫游泳、糖水偏好数据。结果:16周后,与对照组相比,致石组胆汁浑浊,透光性变差,胆囊内可见固体沉淀,致石组胆汁TC水平升高(P<0.05),而TBA、PL水平下降(P<0.05,P<0.05)。“CUMS+孤养”造模期间,CS肝郁组小鼠喜蜷缩于笼子角落,精神欠佳,摄食减少;对照组、CS组小鼠体质量不断增加,而CS肝郁组小鼠体质量不断下降,造模结束后,CS肝郁组体质量明显低于其他两组。“CUMS+孤养”造模前,3组小鼠强迫游泳静止时间及糖水消耗率差异无统计学意义(P值均>0.05),造模结束后,对照组、CS组之间的游泳静止时间及糖水消耗率差异无统计学意义(P值均>0.05),但对比CS组,CS肝郁组小鼠糖水消耗率显著降低(P<0.05)、强迫游泳静止不动时间延长(P<0.05)。结论:高脂致石饲料联合21天“CUMS+孤养”模式可成功制备胆石症肝郁证动物模型。

Objective:To establish an animal model of cholelithiasis with liver stagnation syndrome by using high-fat lithogenic diet combined with 21 day “chronic unpredictable mild stimulation(CUMS)+orphan rearing” mode.Methods:33 male ICR mice were randomly divided into 12 in the control group and 21 in the lithotripsy group.After 16 weeks of feeding with different diets, 3 animals in each group were selected to examine the formation of stones in the gallbladder, bile turbidity, and light transmittance;enzyme-linked immunosorbent assay(ELISA) was used to detect total cholesterol(TC),and total bile acid(TBA) in gallbladder bile, and phospholipid(PL) content to determine whether to establish a cholelithiasis disease model, After the model was successful, the lithotripsy group was randomly divided into a model group(CS group) and a model liver depression group(CS liver depression group).In the CS liver depression group, a “21 day CUMS+orphan rearing” model was used to establish a liver depression model.During the modeling period, the mental status, food intake, behavioral activities of the mice were dynamically observed, and the data of the mices body weight, forced swimming, and sugar water preference were recorded in time.Results:After 16 weeks, compared with the control group, the bile in the lithotripsy group was cloudy, the light transmittance became poorer, and solid precipitation was seen in the gallbladder.The bile TC level in the lithotripsy group increased(P<0.05),while the levels of TBA and PL decreased(P<0.05,P<0.05).During the modeling period of “CUMS+orphan rearing”,the mice in the CS group with liver depression liked to curl up in the corner of the cage, with poor energy and reduced food intake.After modeling, the body weight of CS liver depression group was significantly lower than that of the other two groups.Before the “CUMS+orphan rearing” model, there was no significant difference in the static time of forced swimming and the consumption rate of sugar water among the three groups(all P>0.05).There was no significant difference in the consumption rate of sugar and water(P>0.05),but compared with the CS group, the consumption rate of sugar and water in the CS group was significantly lower(P<0.05),and the immobility time of forced swimming was prolonged(P<0.05).Conclusion:High-fat lithogenic diet combined with 21 day “CUMS+orphan rearing” mode can successfully establish an animal model of cholelithiasis with liver stagnation syndrome.