附子理中丸灌胃后脾阳虚证大鼠海马组织β-EP、cAMP、PKA、神经元细胞μ受体表达观察  

Expression of β-EP,cAMP,PKA,and μ-receptor in hippocampus of rats with spleen Yang deficiency syndrome after administration of Fuzi Lizhong pills

作　　者：孙大宇[1] 单德红[1] 刘旭东[1] 刘文俊[1] 王德山[1] SUN Dayu;SHAN Dehong;LIU Xudong;LIU Wenjun;WANG Deshan(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)

机构地区：[1]辽宁中医药大学生理教研室,沈阳110847

出　　处：《山东医药》2018年第43期6-9,共4页Shandong Medical Journal

基　　金：国家重点基础研究发展计划(973计划)资助项目(2013CB531702)

摘　　要：目的观察附子理中丸灌胃的脾阳虚证模型大鼠海马组织β内啡肽(β-EP)、环磷酸腺苷(c AMP)、蛋白激酶A(PKA)及神经元细胞μ受体的表达变化。方法 40只SPF级SD雄性大鼠随机分为空白组、模型组、中药组、自愈组,每组10只。模型组、中药组、自愈组采用饮食失节+劳倦等复合因素方法制作脾气虚模型,加用"苦寒泻下"法(连续灌服100%番泻叶)制作脾阳虚模型。空白组不做任何处理。造模后中药组予附子理中丸2 m L/(100 g·d)灌胃,1次/d,给药2周。(1)大鼠行为运动功能观察采用旷场实验检测各组大鼠行为学改变(水平运动距离、直立次数)采用YLS-13A大小鼠抓力测定仪检测各组大鼠前肢抓力。(2)分别于第2、4周处死各组大鼠,取海马组织,ELISA法检测海马组织β-EP、c AMP、PKA,免疫组化SABC法检测神经元细胞μ受体。结果与空白组比较,模型组、自愈组大鼠水平运动距离和直立次数均降低(P均<0. 01)。与空白组比较,模型组大鼠前肢抓力下降(P <0. 01);与模型组比较,中药组和自愈组大鼠前肢抓力增强(P均<0. 01)。与空白组比较,模型组及自愈组海马组织中β-EP含量升高、c AMP和PKA含量降低(P均<0. 05);与模型组比较,中药组c AMP和PKA含量升高,自愈组PKA含量升高(P均<0. 05);与自愈组比较,中药组海马组织中β-EP含量降低、c AMP和PKA含量升高(P均<0. 05)。与空白组比较,模型组、自愈组大鼠海马组织μ受体光密度升高(P均<0. 05);与模型组比较,中药组大鼠海马组织中神经元细胞μ受体光密度降低(P <0. 05);与自愈组比较,中药组大鼠海马组织中神经元细胞μ受体光密度降低(P <0. 05)。结论附子理中丸可改善脾阳虚证模型大鼠的脾阳虚证"神疲、乏力"表征,其机制可能降低海马组织β-EP含量、升高c AMP及PKA含量、增加神经元细胞μ受体表达。Objective To observe the changes in the expression ofβ-endorphin(β-EP),cyclic adenosine monophosphate(cAMP),protein kinase A(PKA)andμ-receptor in the hippocampus of rats with spleen Yang deficiency syndrome after treatment of Fuzi Lizhong pills.Methods Forty SPF grade SD male rats were randomly divided into the blank group,model group,traditional Chinese medicine group,and self-healing group,with 10 rats in each group.The model group,the traditional Chinese medicine group and the self-healing group adopted the compound factors such as diet incontinence and fatigue to make the models of spleen deficiency.On this basis,we used the method of diarrhea of bitter cold(continuous irrigation of 100%senna leaves)to make the models of spleen Yang deficiency.The blank group was not treated.Two mL/(100 god)of Fuzi Lizhong pills was given to the rats in the traditional Chinese medicine group after model making,for 2 weeks.In this study,the behavioral and motor functions of rats were observed using the Open field experiment to detect the changes of rats in each group(such as horizontal movement distance and number of upright times).Rats were sacrificed at week 2 and 4,respectively,and hippocampal tissue was taken.Hippocampalβ-EP,cAMP,and PKA were detected by ELISA,andμ-receptor was detected by immunohistochemical SABC.Results Compared with the blank group,the distance of horizontal movement and the number of upright position decreased in the model group and the self-healing group(both P<0.01).Compared with the blank group,the rats in the model group had a decrease in their foreleg grasp(P<0.01).Compared with the model group,the rats in the traditional Chinese medicine group and the self-healing group showed increased grasp strength(P<0.01).Compared with the blank group,the content of the hippocampalβ-EP in the model group and the self-healing group increased and the content of cAMP and PKA decreased(all P<0.05).Compared with the model group,the content of cAMP and PKA in the traditional Chinese medicine group increased,

关 键 词：附子理中丸 脾阳虚证 海马 Β内啡肽 Μ受体 环磷酸腺苷 蛋白激酶A 

分 类 号：R2031[医药卫生—中医学]