电针“足三里”穴对脾气虚大鼠小肽转运体表达的影响

Effect of Electroacupuncture at "Zusanli" on PepT1 Expression in Rats with Deficiency of Spleen Qi

目的:探讨电针"足三里"穴对脾气虚模型大鼠小肠黏膜组织内的小肽转运体(PepT1)的蛋白及基因表达的影响。方法:将40只SD雄性大鼠按照随机数字表法随机分为正常组、脾气虚组、足三里组、非经非穴组,每组10只。大鼠均统一饲养于SPF级动物实验中心。正常组给予正常饮食,其余3组均采用不规则饮食和劳倦过度复合法建立脾气虚证大鼠模型。造模成功之后,对足三里组进行电针"足三里"穴、非经非穴组大鼠电针非经非穴点干预处理7 d。观察各组大鼠免疫器官胸腺指数及脾脏指数的变化;观测各组大鼠小肠推进率;采用HE染色法观察各组大鼠小肠黏膜组织形态的变化;采用蛋白质免疫印迹法检测大鼠小肠黏膜组织内的PepT1的蛋白含量变化;采用荧光定量PCR法检测大鼠小肠黏膜组织的PepT1的mRNA表达水平。结果:脾虚组大鼠的胸腺和脾脏质量以及小肠推进率都明显低于正常组,足三里组相比于脾虚组有所升高;HE染色结果示:与正常组相比,脾气虚组大鼠小肠黏膜组织有萎缩、糜烂等改变,足三里组的组织形态较脾气虚组大鼠相比有所恢复;WB及PCR结果示:与正常组相比,脾气虚组大鼠小肠黏膜组织内的PepT1蛋白和基因表达水平均明显低于正常组(P<0.05);与脾气虚组相比,足三里组大鼠小肠黏膜组织内的PepT1蛋白和基因表达水平有所升高(P<0.05),非经非穴组未见显著性差异(P>0.05)。结论:电针"足三里"穴可以调控脾气虚大鼠小肠黏膜组织内的PepT1蛋白及基因的异常表达,参与小肠对蛋白质的吸收作用,进而改善脾气虚证。

Objective:To investigate the effect of electroacupuncture at Zusanli(ST36)on protein and gene expression of small peptide transporter(PepT1)in small intestinal mucosa of rats with deficiency of spleen Qi.Methods:Forty male SD rats were randomly divided into normal group,spleen Qi deficiency group,Zusanli(ST36)group and non-meridian non-acupoint group.All rats were fed in SPF level animal experimental center.The normal diet was given to the normal group,and the other three groups were treated with irregular diet and overfatigue combined method to establish the model of spleen Qi deficiency syndrome in rats.After the model was established successfully,the Zusanli(ST36)group was treated with electroacupuncture at Zusanli(ST36),and the rats of non-meridian non-acupoint group were treated with non-meridian and non-acupoint intervention for 7 days.We observed the changes of thymus index and spleen index of rats in each group,the propelling rate of small intestine in each group by HE staining.The protein content of PepT1 in small intestinal mucosa of rats was detected by Western blotand the mRNA expression of PepT1 in small intestinal mucosa of rats was detected by fluorescence quantitative PCR method.Results:The weight of thymus and spleen and the rate of small intestine propulsion in spleen deficiency group were significantly lower than those in normal group.The results of HE staining in Zusanli(ST36)group compared with spleen deficiency group showed that the histological morphology of Zusanli(ST36)group was better than that of spleen Qi deficiency group.Compared with the normal group,the levels of PEPT1 protein and gene expression in the small intestinal mucosa of the spleen Qi deficiency group were significantly lower than those of the normal group(P<0.05),and those of the spleen Qi deficiency group were significantly lower than those of the spleen Qi deficiency group.In Zusanli(ST36)group,the expression of PEPT1 protein and gene in small intestinal mucosa was increased(P<0.05),but no significant difference was found in non-acupoint group(P<0.05).Conclusion:Electroacupuncture at Zusanli(ST36)can regulate the abnormal expressions of PEPT1 protein and gene in the small intestinal mucosa of spleen Qi deficiency rats,participate in the absorption of protein by the small intestine,and then improve the spleen Qi deficiency syndrome.