琥珀酸曲格列汀对糖尿病模型小鼠胆汁酸代谢的影响

Effects of Trelagliptin Succinate on Bile Acid Metabolism in Diabetes Mellitus Model Mice

目的 探讨琥珀酸曲格列汀对2型糖尿病(T2DM)胆汁酸代谢的影响。方法 将18只db/db小鼠随机分成模型对照组、琥珀酸曲格列汀组、磷酸西格列汀组,每组6只,C57小鼠6只为对照组。琥珀酸曲格列汀组给药剂量为17.29 mg·kg^(-1),每周1次,磷酸西格列汀组剂量为16.71mg·kg^(-1)·d^(-1),连续给药8周。苏木精-伊红(HE)染色观察各组小鼠的胰腺和肾脏组织病理状态;Real-time PCR和Western blotting法测定小鼠肝脏CYP7A1、法呢醇X受体(FXR)和小异二聚体伴侣(SHP)的mRNA和蛋白表达水平;基于靶向代谢组学检测小鼠回肠内容物中69种胆汁酸的含量。结果 与模型对照组比较,琥珀酸曲格列汀组和磷酸西格列汀组胰腺和肾脏组织病理状态改善;琥珀酸曲格列汀组和磷酸西格列汀组CYP7A1 mRNA和蛋白表达水平差异无统计学意义(P>0.05);琥珀酸曲格列汀组FXR mRNA和蛋白表达水平显著升高(P<0.05或P<0.01),而磷酸西格列汀组FXR mRNA和蛋白表达水平无明显变化(P>0.05);琥珀酸曲格列汀组SHP mRNA和蛋白表达水平有升高趋势,但差异无统计学意义(P>0.05),磷酸西格列汀组对SHP mRNA和蛋白表达水平差异无统计学意义(P>0.05);琥珀酸曲格列汀组去甲胆酸(NCA)、猪脱氧胆酸(HDCA)、牛磺猪去氧胆酸(THDCA)和α-鼠胆酸(αMCA)显著下调,牛磺鹅去氧胆酸(TCDCA)显著上调;磷酸西格列汀组NCA、HDCA、αMCA、THDCA、牛磺脱氧胆酸(TDCA)和牛磺α-鼠胆酸(TαMCA)显著下调。结论 琥珀酸曲格列汀可增加db/db小鼠FXR和SHP的表达,可能是因为上调TCDCA并下调了NCA和αMCA。

Objective To explore the effect of trelagliptin succinate on bile acid metabolism in type 2 diabetes mellitus(T2DM).Methods Eighteen db/db mice were randomly divided into model control group(Mod),trelagliptin succinate group(Tre)and sitagliptin phosphate group(Sit)(n=6),and 6 C57 mice were in control group(Con).The doses of trelagliptin succinate and sitagliptin phosphate were 17.29 mg·kg^(-1)·w^(-1) and 16.71 mg·kg^(-1)·d^(-1),respectively,and were administered continuously for eight weeks.HE staining was used to observe the pathological status of the pancreas and kidney of the mice in each group;Real-time PCR and Western Blot assay were used to determine the mRNA and protein expression levels of CYP7A1,FXR and SHP in the liver of the mice;the content of bile acids in mouse ileum was detected based on targeted metabolomics.Results Compared with the Mod group,the pathological state of the pancreas and kidney were improved in the Tre group and the Sit group.The Tre group and the Sit group had no significant difference in the expression levels of CYP7A1 mRNA and protein(P>0.05);the expression levels of FXR mRNA and protein in Tre group were significantly increased(the former P<0.05,the latter P<0.01),while sitagliptin had no significant effect on the expression levels of the FXR mRNA and protein in Sit group(P>0.05);the Tre group showed a significant effect on expression levels of the SHP mRNA and protein,but there was no significant difference(P>0.05),and Sit group showed no significant difference in expression levels of the SHP mRNA and protein(P>0.05);the Tre could significantly down-regulate NCA,HDCA andαMCA,and significantly up-regulated TCDCA;Sit could significantly down-regulate NCA,HDCA,αMCA,THDCA,TDCA and TαMCA.Conclusion Trelagliptin succinate can increase the expression levels of FXR and SHP in db/db mice,possibly due to the up-regulation of TCDCA and the down-regulation of NCA andαMCA.