摘要
目的分析脾虚模型大鼠脑肠肽〔β-内啡肽(β-EP)、胆囊收缩素(CCK)、血管活性肽(VIP)〕、下丘脑葡萄糖转运体(GLUT)1、GLUT3表达水平变化。方法 2015年3—9月,采用随机数字表法将24只SPF级雄性SD大鼠分为对照组、脾气虚组、脾阳虚组,每组8只。脾气虚模型的建立采用饮食失节结合劳倦过度的原则,脾阳虚模型则是在脾气虚的基础上施加苦寒泻下法完成。观察3组大鼠一般情况(体征状态、体质量、体温、进食量等)及前肢抓力。采用酶联免疫吸附试验(ELISA)法测定下丘脑、胃、空肠β-EP、CCK、VIP表达水平,实时荧光定量聚合酶链式反应(Real-time PCR)法测定下丘脑GLUT1、GLUT3 mRNA表达水平,Western blotting法测定下丘脑GLUT1、GLUT3表达水平。结果脾气虚组大鼠体质量、进食量、前肢抓力低于对照组(P<0.05);脾阳虚组大鼠体质量、体温、进食量、前肢抓力低于对照组(P<0.05);脾阳虚组大鼠体温低于脾气虚组(P<0.05)。脾气虚组大鼠下丘脑β-EP表达水平低于对照组,胃、空肠β-EP表达水平及下丘脑、胃、空肠CCK、VIP表达水平高于对照组(P<0.05);脾阳虚组大鼠下丘脑β-EP表达水平低于对照组,胃、空肠β-EP、CCK表达水平及下丘脑、胃、空肠VIP表达水平高于对照组(P<0.05);脾阳虚组大鼠下丘脑β-EP表达水平及下丘脑、空肠CCK表达水平低于脾气虚组,胃、空肠β-EP表达水平及下丘脑、胃、空肠VIP表达水平高于脾气虚组(P<0.05)。脾气虚组、脾阳虚组大鼠下丘脑GLUT1、GLUT3 mRNA及其蛋白表达水平低于对照组(P<0.05);脾阳虚组大鼠下丘脑GLUT1、GLUT3 mRNA及其蛋白表达水平低于脾气虚组(P<0.05)。结论脾虚状态下,大鼠下丘脑GLUT1、GLUT3 mRNA及其蛋白表达水平下降,导致下丘脑、胃、空肠β-EP、CCK、VIP表达水平异常,这可能是脾虚本质研究的又一新观点。
Objective Analysis the changes of the expression levels of brain - gut peptide〔β - endorplhin(β-EP), cholecystokinin(CCK)and vasoactive peptide( VIP)〕and glucose transporter - 1( GLUT1)and glucose transporter - 3 (GLUT3)in hypothalamus of spleen deficiency rats. Methods From March to September 2015,a total of 24 SPF male SD rats were divided into control group,spleen - qi deficiency group and spleen - yang deficiency group with 8 rats in each group by random number table method. The model rats of spleen - qi deficiency were built by immoderate diet and overfatigue;the model rats of spleen - yang deficiency were built by bitter and cold aloes on the basis of the model rats of spleen - qi deficiency. The three groups were observed by general information( signs and state,body mass,temperature and food - intake)and grip of forelegs. The expression levels of β-EP,CCK and VIP in hypothalamus,stomach and jejunum were assessed by enzyme linked immunosorbent assay(ELISA);the mRNA levels of GLUT1 and GLUT3 were measured by real time quantitative fluorescent&amp;nbsp;polymerase chain reaction( Real - time PCR);the levels of GLUT1 and GLUT3 in hypothalamus were measured by Western blotting. Results The body mass,food - intake and grip of forelegs of spleen - qi deficiency group were lower than those of control group(P 〈 0. 05);the body mass,temperature,food - intake and grip of forelegs of spleen - yang deficiency group were lower than those of control group(P 〈 0. 05);the temperature of spleen - yang deficiency group was lower than that of spleen -qi deficiency group(P 〈 0. 05). The expression level of β-EP in hypothalamus of spleen - qi deficiency group was lower than that of control group(P 〈 0. 05);the expression level of β-EP in stomach and jejunum and the expression levels of CCK and VIP in hypothalamus,stomach and jejunum of spleen - qi deficiency group were higher than those of control group(P 〈 0. 05). The expression level of β-EP in hypothalamus of spleen - yang deficiency group was lower than that of control group and the expression levels of β-EP and CCK in stomach and jejunum and the expression level of VIP in hypothalamus,stomach and jejunum of spleen- yang deficiency group were higher than those of control group(P 〈 0. 05);the expression level of β-EP in hypothalamus and the expression level of CCK in hypothalamus and jejunum of spleen - yang deficiency group were lower than those of spleen - qi deficiency group and the expression level of β-EP in stomach and jejunum and the expression level of VIP in hypothalamus, stomach and jejunum of spleen - yang deficiency group were higher than those of spleen - qi deficiency group(P 〈 0. 05). The mRNA expression and protein expression of GLUT1 and GLUT3 in hypothalamus of spleen - qi deficiency group and spleen - yang deficiency group were lower than those of control group(P 〈 0. 05);the mRNA expression and protein expression of GLUT1 and GLUT3 in hypothalamus of spleen - yang deficiency group were lower than those of spleen - qi deficiency group(P 〈 0. 05). Conclusion Under the condition of spleen deficiency,the mRNA expression levels of GLUT1 and GLUT3 in hypothalamus are lower and the expression levels of β-EP,CCK and VIP in hypothalamus,stomach and jejunum are abnormal,which may be a new viewpoint of studying the essence of spleen deficiency.
出处
《中国全科医学》
CAS
CSCD
北大核心
2016年第18期2201-2205,共5页
Chinese General Practice
基金
国家重点基础研究发展计划(973计划)课题(2013CB531702)
关键词
脾气虚
脾阳虚
脑肠肽
葡萄糖转运蛋白质
下丘脑
Spleen - qi deficiency
Spleen - yang deficiency
Brain - gut peptide
Glucose transport protein
Hypothalamus