基于饮食劳倦伤脾理论的五种脾气虚证造模方法的比较

Comparative Study on Five Modeling Methods of Spleen Qi Deficiency Syndrome Based on the Theory that“Diet and Fatigue Damage the Spleen”

目的比较五种常见的脾气虚证造模方法,以建立符合中医临床的脾气虚证动物模型。方法60只雄性昆明小鼠随机分为正常组、饮食失节组、跑步组、站立组、跑步+饮食失节组、站立+饮食失节组,每组10只。饮食失节组单日禁食不禁水,双日自由饮食饮水;跑步组以16 m/min的速度每天跑步20 min;站立组在自制水环境小平台箱内站立每天4 h;跑步+饮食失节组以16 m/min的速度每天跑步20 min,同时单日禁食不禁水,双日自由饮食饮水;站立+饮食失节组在自制水环境小平台箱内站立每天4 h,联合单日禁食不禁水,双日自由饮食饮水。各组持续造模14天。观察各组小鼠一般情况,隔日记录小鼠的摄食量、饮水量,计算粪便含水率;旷场实验观察小鼠5 min运动距离、平均速度;计算胃残留率、小肠推进率,测定肠道消化酶活性、血清中乳酸脱氢酶(LDH)含量,评价脾气虚模型。测定血清中sigA含量、血常规、脏器指数,评价免疫功能。结果与正常组相比,饮食失节组、跑步+饮食失节组小鼠暴躁易怒,被毛无光泽,粪便颗粒较小、无光泽、硬而干,粪便含水率呈下降趋势,每日摄食量、饮水量呈增长趋势;跑步组小鼠反应灵敏,被毛无光泽,粪便软硬适中,粪便含水率、饮水量呈增长趋势,每日摄食量呈下降趋势;站立组小鼠反应迟钝、眯眼、弓背,被毛蓬枯槁无光泽,粪便呈条形软便,粪便含水率呈增长趋势,每日摄食量、饮水量呈下降趋势;站立+饮食失节组小鼠暴躁易怒,被毛无光泽,粪便软硬适中,粪便含水率、每日摄食量、饮水量整体表现出增长趋势。与正常组比较,饮食失节组、跑步组小鼠5 min运动距离、平均速度,跑步组脾脏指数显著升高(P<0.05)。站立组小鼠5 min运动距离、平均速度显著低于其他造模组,胃残留率显著低于饮食失节组,小肠推进率显著高于正常组、饮食失节组、跑步组、跑步+饮食失节组,小肠黏膜蛋白酶及蔗糖酶活性显著低于正常组、站立+饮食失节组,肝脏指数显著高于跑步组(P<0.05)。站立组、站立+饮食失节组小鼠血清中LDH含量显著低于饮食失节组(P<0.05)。站立+饮食失节组肝脏指数显著高于其余各组,血小板数显著低于正常组(P<0.05)。饮食失节组肝脏指数显著高于正常组、跑步组、跑步+饮食失节组,血红蛋白数显著低于站立组(P<0.05)。跑步+饮食失节组红细胞数显著低于站立组、站立+饮食失节组,血红蛋白数显著低于正常组、站立组、站立+饮食失节组(P<0.05)。结论利用水环境小平台建立的小鼠模型更符合脾气虚证诊断标准。

Objective To compare the five modeling methods of spleen qi deficiency syndrome,and to establish animal models consistent with clinical spleen qi deficiency in traditional Chinese medicine(TCM).Methods Sixty male Kunming mice were randomly divided into normal group,diet disorder group,running group,standing group,running with diet disorder group,and standing with diet disorder group,ten mice in each group.The diet disorder group was fasted but with water for one day,and free to eat and drink for the next day.The running group ran at a speed of 16 m/min for 20 minutes every day.The standing group stood in the self-made small water environment platform box for 4 hours every day.The running with diet disorder group and the standing with diet disorder group each received the corresponding combined two interventions.All groups continued modeling for 14 days.The general condition of the mice was observed;food intake and water intake were recorded every other day,and the moisture content of feces was calculated.Open-field experiment was used to evaluate the five minute walking distance and average speed of mice.The gastric residual rate and small intestinal propulsion rate were calculated.The intestinal digestive enzyme activity and serum lactate dehydrogenase(LDH)content were detected.The serum sigA content,blood routine indicators,organ index and immune function were measured.Results Compared to the normal group,the mice in the normal group were responsive,with shiny coats,wheat-like feces,moderate softness and hardness,and an increasing trend of food intake.Compared to the normal group,the mice in the diet disorder and the running+diet disorder groups were irritable,with a lustrous coat,dull,hard and dry feces in small pieces,having decreased moisture content of feces and increased amount of food and water intake;the mice in the running group were responsive,with lusterless coats,moderately soft and hard feces,having increased fecal moisture content and food intake and decreased water intake;the mice in the standing group were unresponsive,squinting,arching their backs,with lusterless coats,soft stools in the shape of strips,easily deformed when picked up with forceps,increased moisture content of feces,and decreased food and water intake;the mice in the standing+diet disorder group were irritable,with lusterless coat and moderate fecal hardness,as well as increased fecal moisture content,daily food and water intake.The five minute distance and average speed of mice in the diet disorder group and the running group,as well as the spleen index in the running group were significantly higher than those in the normal group(P<0.05).The standing group had significantly decreased five minute distance and average speed than the diet disorder group,the running group,the running+diet disorder group,and the standing+diet disorder group,lowered gastric residual rate than the diet disorder group,increased small intestine propulsion rate than the normal,diet disorder,running,running+diet disorder groups,lowered small intestine mucosal protease and sucrase activity than the normal and stand+diet disorder group,as well as increased liver index than the running group(P<0.05).And the serum LDH content of mice in the standing group and the standing+diet disorder group was significantly lower than that in the diet disorder group(P<0.05).The standing+diet disorder group had higher liver index than the other groups,and lower platelet count than the normal group(P<0.05).The diet disorder group had higher liver index than the normal,running,and running+diet disorder groups,and lower hemoglobin count than the standing group(P<0.05).The running+diet disorder group had decreased red blood cells than the standing and standing+diet disorder groups,and decreased hemoglobin count than the normal,standing,and standing+diet disorder groups(P<0.05).Conclusion The model established by using the small platform of the water environment is more consistent with the diagnostic criteria of spleen qi deficiency syndrome.