三种气虚证动物模型的比较与评价

Comparison and Evaluation of Three Animal Models of Qi Deficiency Syndrome

目的比较环磷酰胺、限食+力竭游泳、限食+力竭游泳+环磷酰胺三种方法制备的气虚证实验动物模型,确定最优造模方法。方法将美国癌症研究所(ICR)小鼠按力竭游泳时间随机分为正常对照组、环磷酰胺组、限食+力竭游泳组、限食+力竭游泳+环磷酰胺组,每组10只,雌雄各半。造模时间共15天。环磷酰胺组小鼠于造模第1、2、3、10天腹腔注射环磷酰胺,每次50 mg/kg,共4次;限食+力竭游泳组小鼠连续15天每天进行力竭游泳,并限制进食量为150 g/(kg·d);限食+力竭游泳+环磷酰胺组小鼠按上述方法进行力竭游泳、限制进食量、腹腔注射环磷酰胺。每日观察小鼠一般状态,于造模第7、14天测定小鼠体质量,造模15天后比较各组小鼠力竭游泳时间、脾指数、胸腺指数、肝糖原含量,血浆尿素氮、血浆免疫球蛋白IgG和IgA含量,以及外周血T淋巴细胞(CD3^(+))及其亚群(CD4^(+)、CD8^(+))、B淋巴细胞(CD19^(+))含量。结果与正常对照组比较,环磷酰胺组小鼠体质量、力竭游泳时间、脏器指数、外周血CD19+含量、血浆免疫球蛋白IgG、IgA含量显著降低,血浆尿素氮含量显著升高(P<0.05或P<0.01);限食+力竭游泳组小鼠体质量,脏器指数,外周血CD3^(+)、CD4^(+)、CD19^(+)及肝糖原含量显著降低(P<0.05或P<0.01);限食+力竭游泳+环磷酰胺组小鼠体质量,力竭游泳时间,脏器指数,外周血CD3^(+)、CD4^(+)、CD8^(+)、CD19^(+),肝糖原,血浆免疫球蛋白IgG、IgA含量显著降低(P<0.05或P<0.01)。与环磷酰胺组比较,限食+力竭游泳组、限食+力竭游泳+环磷酰胺组小鼠体质量、胸腺指数、外周血CD4^(+)、CD19^(+)、肝糖原含量明显降低,且限食+力竭游泳组IgG显著升高(P<0.05)。与限食+力竭游泳组比较,环磷酰胺组脾指数显著下降;限食+力竭游泳+环磷酰胺组小鼠脾指数、CD19^(+)、血浆中IgG含量明显降低(P<0.05)。结论三种模型均可从免疫、能量、代谢不同方面模拟气虚证的临床表现,且力竭游泳+限食+环磷酰胺造模方法能更好地模拟气虚证“神疲乏力”的临床表现。

Objective To compare and evaluate three experimental animal models of qi deficiency syndrome prepared by cyclophosphamide,food restriction plus exhaustive swimming,and cyclophosphamide plus food restriction plus exhaustive swimming,and to determine the optimal modeling method.Methods The Institute of Cancer Research(ICR)mice were divided into normal control group,cyclophosphamide model group,food restriction plus exhaustive swimming model group,food restriction plus exhaustive swimming plus cyclophosphamide model group,with 10 mice in each group,half-female and half-male.The modelling time was 15 days.Cyclophosphamide was intraperitoneally injected into the mice on the 1st,2nd,3rd,and 10th days of modelling(50 mg/kg).The mice in the exhaustive swimming plus food restriction model group performed exhaustive swimming every day for 15 days,and the food intake was restricted into 150 g/(kg·d).The mice in the exhaustive swimming plus food restriction plus cyclophosphamide model group performed exhaustive swimming,restricted food intake,and intraperitoneal injection of cyclophosphamide as mentioned above.The general condition of the mice was observed every day,and the body weight was measured on the 7th and 14th day of modelling.The exhaustive swimming time,splenic index,thymus index,liver glycogen content,plasma urea nitrogen,plasma immunoglobulin Ig G and Ig A content,as well as peripheral blood T lymphocytes(CD3^(+))and their subsets(CD4^(+),CD8^(+)),and B lymphocyte(CD19^(+))content were detected and compared between groups after 15-day modelling.Results The body weight,exhaustive swimming time,organ index,peripheral blood CD19^(+)content,plasma Ig G and Ig A content of mice in cyclophosphamide group significantly decreased,while plasma urea nitrogen content significantly increased compared tho those of mice in the normal control group(P<0.05 or P<0.01).The body weight,organ index,peripheral blood CD3^(+),CD4^(+),CD19^(+)and liver glycogen of mice in the exhaustive swimming plus food restriction group decreased(P<0.05 or P<0.01),and also the body weight,exhaustive swimming time,organ index,peripheral blood CD3^(+),CD4^(+),CD8^(+),CD19^(+),liver glycogen content,plasma immunoglobulin Ig G and Ig A content in exhaustive swimming plus food restriction plus cyclophosphamide group significantly decreased(P<0.05 or P<0.01)compared to those in the normal control group.Compared to those in the cyclophosphamide group,the body weight,thymus index,peripheral blood CD4^(+),CD19^(+)content,and liver glycogen content in the exhaustive swimming plus food restriction group and the exhaustive swimming plus food restriction plus cyclophosphamide group were significantly reduced,while the Ig G content in the exhaustive swimming plus food restriction group significantly increased(P<0.05).Compared to those in the exhaustive swimming plus food restriction group,the splenic index in the cyclophosphamide group significantly decreased,while splenic index,CD19^(+)content,and plasma immunoglobulin Ig G in the exhaustive swimming plus food restriction plus cyclophosphamide group were significantly reduced(P<0.05).Conclusion The three models could all simulate the clinical manifestations of qi deficiency syndrome from the aspects of immunity,energy and metabolism,and the exhaustive swimming plus food restriction plus cyclophosphamide modeling method is more in line with the mental fatigue symptom of qi deficiency syndrome.