| Purpose:To establish a model rat with spleen-qi deficiency,and to treat the expression of the related transporter protein SGLT1,GLUT2 and Pept T1 at the gene level and protein level in the small intestine mucosal epithelial tissue of rats with acupuncture thorn "Zusanli"(ST36)and the stimulation of electro acupuncture.The changes of the expression of various indexes in vivo before and after the experiment were aimed at discussing the effect of electro acupuncture on the absorption function of small intestine on nutrients such as sugars and proteins,and then revealing the modern scientific connotation of "spleen main transport" from the molecular biology level.Methods:The experiment was conducted in the Laboratory of Research platform and Animal experiment Center of Liaoning University of traditional Chinese Medicine in May2017 and May 2018.40 SD Male rats were randomly divided into normal group,spleen-qi deficiency Group,“Zusanli”(ST36)group,non-acupuncture point Group,10 in each group.All the rats were raised at the SPF Level Animal Experiment Center.In addition to the normal group,the remaining 3 groups used fatigue excessive and irregular diet compound method to establish a model of spleen-qi deficiency syndrome.After the success of the mold,“Zusanli”(ST36)group,non-acupuncture point group of rats were respectively stimulated by electro acupuncture "Zusanli"(ST36)acupoint,non-acupuncture point,continuous intervention treatment for 7 days.During the whole process,the body weight of rats was monitored,the thymic index and spleen index of rats in each group were measured,and the propulsive rate of small intestine in each group was measured.The morphological changes of small intestine mucosal tissue in each group of rats were observed by HE staining,and the m RNA expression levels of glucose transporter SGLT1,GLUT2 and small peptide transporter Pep T1 in rat small intestine mucosal epithelium were detected by fluorescence quantitative PCR,and the real-time was used by??CT method.The results of PCR were quantitatively analyzed,and the protein content of glucose transporter SGLT1,GLUT2 and small peptide transporter Pept1 in rat small intestine mucosal epithelial tissue was detected by Western Blot method(WB).Results:1.After electro acupuncture on both sides of "Zusanli"(ST36)rats,the levels of spleen index,thymus index and small intestine propulsive rate were all increased.2.Morphological changes of intestinal mucosa after electro acupuncture on both sides of Zusanli in rats with spleen-qi deficiency.HE staining shows:The intestinal mucosal tissue of Zusanli group was recovered to a certain extent after partial injury of small intestine mucosa in the spleen-qi deficiency group.3.Changes of glucose transporter protein and gene expression in intestinal mucosa after electro acupuncture of Zusanli on both sides of spleen-qi deficiency rats.The expression levels of glucose transporter SGLT1 and GLUT2 in intestinal mucosa of spleen-qi deficiency group and non-meridian non-acupoint group were significantly lower than those of normal group(P<0.05).The expression of SGLT1 and GLUT2 protein in intestinal mucosa of Zusanli group was higher than that of spleen-qi deficiency group(P<0.05),but there was no significant difference in non-acupuncture point group(P>0.05).The gene expression levels of glucose transporter SGLT1 and GLUT2 in intestinal mucosa of rats in spleen-qi deficiency group and non-meridian non-acupuncture point group were significantly lower than those in normal group(P<0.05).The expression level of SGLT1 and GLUT2 in intestinal mucosa of Zusanli group was higher than that of spleen-qi deficiency group(P<0.05),but there was no significant difference in non-acupuncture point group(P>0.05).4.Changes of small peptide transporter protein and gene expression in small intestinal mucosa after electro acupuncture on both sides of Zusanli in rats with spleen-qi deficiency.The protein expression level of small peptide transporter(Pep T1)in small intestine mucosa of spleen-qi deficiency group and non-acupuncture point group was significantly lower than that of normal group(P<0.05).The expression of Pep T1 protein in intestinal mucosa of rats in Zusanli group was higher than that in spleen-qi deficiency group(P<0.05),but there was no significant difference in non-acupuncture point group(P>0.05).The gene expression level of small peptide transporter(Pep T1)in small intestine mucosa of spleen-qi deficiency group and non-meridian non-acupuncture point group was significantly lower than that of normal group(P<0.05).The expression of Pep T1 gene in intestinal mucosa of rats in Zusanli group was higher than that in spleen-qi deficiency group(P<0.05),but there was no significant difference in non-acupuncture point group(P>0.05).Conclusion:1.Electro acupuncture of Zusanli(ST36)can repair intestinal mucosal injury and improve gastrointestinal motility in spleen-deficiency rats.2.Electro acupuncture stimulation of Zusanli(ST36)can regulate the abnormal expression of SGLT1,GLUT2 and Pep T1 in the intestinal mucosa of rats with spleen deficiency,and regulate the absorption function of the small intestine,so as to play the role of invigorating the spleen and invigorating qi.3.In the non-acupuncture point group,electro acupuncture intervention failed to improve the state of spleen-qi deficiency,which further verified the specificity of the therapeutic effect of Zusanli(ST36)point. |
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