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目的观察理中汤对脾阳虚模型大鼠小肠上皮肽转运载体蛋白1对二肽转运功能改变、肽转运载体PepT1及其mRNA表达变化的影响。方法采用利血平复合因素造模法建立脾阳虚大鼠模型,以头孢氨苄做为PepT1的转运底物,通过反相高效液相法(RP-HPLC)测定其血药浓度来反映脾阳虚大鼠小肠上皮细胞PepT1的吸收转运功能,RT-PCR测定大鼠小肠黏膜PepT1 mRNA表达,免疫组织化学方法观察大鼠小肠PepT1蛋白表达情况,同时用酶联免疫法测定小肠黏膜组织cAMP含量。结果模型组大鼠血清中cefalexin浓度明显高于正常组,经理中汤治疗后,大鼠cefalexin血药浓度与正常对照组比较无显著性差异;模型组大鼠PepT1蛋白表达明显上调(P<0.05),理中汤治疗组大鼠的PepT1蛋白表达与正常组比较未见明显差异;脾阳虚模型大鼠PepT1 mRNA表达水平较正常组明显下调(P<0.05),经理中汤治疗后,其下调趋势被逆转,与正常组比较未见明显差异;模型组大鼠小肠黏膜cAMP浓度较之于正常组,明显降低,经过理中汤治疗恢复接近正常组水平。结论脾阳虚模型大鼠小肠黏膜PepT1转运功能增强,蛋白表达上调,理中汤治疗可逆向调节这种改变。
Objective To observe the effect of Li Zhong Decoction on changes of dipeptide transport function and peptide transporter PepT1 and its mRNA expression in small intestine peptide transporter of spleen - Yang deficiency model rats. Methods Reserpine model rats were used to establish the rat model of spleen yang deficiency, and cephalexin was used as the transport substrate for PepT1. The plasma concentration of PepT1 was determined by reverse-phase high performance liquid chromatography (RP-HPLC) Absorption and transport function of PepT1 in imaginary rat intestinal epithelial cells, PepT1 mRNA expression in rat intestinal mucosa by RT-PCR, PepT1 protein expression in small intestine by immunohistochemical method, and cAMP content in intestinal mucosa by enzyme-linked immunosorbent assay. Results The serum concentration of cefalexin in the model group was significantly higher than that in the normal group. There was no significant difference in the serum concentration of cefalexin between the model group and the normal control group. The PepT1 protein expression in the model group was significantly increased (P <0.05 ). There was no significant difference in the expression of PepT1 protein between the treatment group and the normal control group. The expression of PepT1 mRNA in the spleen-Yang deficiency model group was significantly lower than that in the normal group (P <0.05) The downward trend was reversed, and no significant difference compared with the normal group; the model group of small intestinal mucosa cAMP concentration was significantly lower than the normal group, after the treatment of Li Zhongtang recovery close to the normal level. Conclusion The spleen yang deficiency model rat intestinal PepT1 transport function increased, protein expression increased, Li Zhong Tang treatment can reverse the regulation of this change.