卡格列净对动脉粥样硬化小鼠氨基酸代谢的作用

Effects of canagliflozin on amino acid metabolism in atherosclerotic mice

目的探索钠-葡萄糖共转运蛋白-2抑制剂卡格列净防治动脉粥样硬化(AS)的可能机制。方法使用西方饮食饲养ApoE-/-小鼠并随机分为模型组(n=10)和卡格列净组(n=10);普通饲料喂养C57BL/6 J小鼠作为对照组(n=10)。给予卡格列净组小鼠卡格列净每日灌胃。干预周期为14周。留取心脏组织行主动脉根部HE染色和油红O染色以评价造模情况及AS损伤程度;PCR法检测一氧化氮合酶的基因表达。采用RNA-seq和液相-质谱法分别进行小鼠肝脏转录组学分析和氨基酸靶向检测。结果主动脉根部HE染色和油红O染色显示西方饮食饲养ApoE-/-小鼠14周可成功构建AS模型;卡格列净可减少小鼠主动脉根部斑块面积(P=0.012)。肝脏转录组学分析提示卡格列净对ApoE-/-小鼠氨基酸代谢有影响,尤其是精氨酸生物合成途径。氨基酸靶向检测显示,卡格列净可减少肝脏L-丝氨酸、L-天冬氨酸、酪氨酸、L-羟脯氨酸和L-瓜氨酸含量,且增加了L-精氨酸含量(P均<0.05);与模型组比较,卡格列净组血清精氨酸、一氧化氮水平及主动脉一氧化氮合酶mRNA相对表达量均较高(P均<0.05)。结论卡格列净可调节动脉粥样硬化小鼠氨基酸代谢,减少生糖氨基酸,并促进精氨酸的生物合成。

Objective To explore the possible anti-atherosclerotic mechanisms of glucose co-transporter-2 inhibitor canagliflozin.Methods ApoE-/-mice fed on Western diet were randomly assigned into the model group(n=10)and the canagliflozin group(n=10).C57BL/6J mice fed on normal diet were chosen as the control group(n=10).Mice in the canagliflozin group were gavaged with canagliflozin for 14 weeks.The presence and severity of atherosclerosis were evaluated with HE and oil red O stainings in aortic root section slices.PCR assay was performed to determine the mRNA expression levels of nitric oxide synthase.Hepatic transcriptome analysis and hepatic amino acid detection were conducted using RNA-seq and targeted LC-MS,respectively.Results HE staining and oil red O staining of the aortic root showed that AS models were successfully established in ApoE-/-mice fed on Western diet for 14 weeks.Canagliflozin alleviated the severity of atherosclerosis in pathology.Hepatic transcriptome analysis indicated that canagliflozin impacted on amino acid metabolism,especially arginine synthesis in ApoE-/-mice.Targeted metabolomics analysis of amino acids showed that canagliflozin reduced hepatic levels of L-serine,L-aspartic acid,tyrosine,L-hydroxyproline,and L-citrulline,but raised the hepatic level of L-arginine.Compared to the model group,the canagliflozin group exhibited higher serum arginine and nitric oxide levels as well as elevated nitric oxide mRNA expression in aortic tissues(P<0.05).Conclusion Canagliflozin regulated the amino acid metabolism,reduced the levels of glucogenic amino acids,and promoted the synthesis of arginine in atherosclerotic mice.