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。将细胞接种到96孔板中。然后在37℃下用0.15mL测定缓冲液（137mM NaCl，5mM KCl，1mM CaCl 2,1mM MgCl 2,50mM HEPES，pH 7.4）洗涤细胞一次。除去测定缓冲液后，加入50μL含有5μL卡那利嗪（0.3-300nM）的新鲜测定缓冲液，然后孵育10分钟。然后，将5μL的6mMα-甲基-D-吡喃葡萄糖苷（AMG，选择性SGLT1 \u002F 2底物）\u002F 14C-AMG（0.07μCi）加入细胞中并在37℃下孵育2小时。接下来，用0.15mL冰冷的磷酸盐缓冲盐水（PBS）洗涤细胞3次。吸出最后一次洗涤后，加入50μL微型筛20。该板由TopCount [1]计数。",{"key":248,"value":249},"动物实验","小鼠和大鼠[1]在这些实验中使用四种啮齿动物模型：（1）喂食高脂肪饮食的雄性C57BL \u002F 6J小鼠（D-12492，含60kcal％脂肪）（饮食诱导的肥胖，胰岛素抵抗小鼠[DIO] ]）; （2）雄性C57BL \u002F ksj-db \u002F db高血糖小鼠; （3）男性Zucker脂肪（ZF）肥胖，胰岛素抵抗大鼠; （4）雄性ZDF肥胖，高血糖大鼠。为了检查Canagliflozin对高血糖症的影响，将单剂量的Canagliflozin（0.1,1和10mg \u002F kg）给予过夜禁食的db \u002F db小鼠。在给药后0,0.5,1,3,6和24小时监测BG水平。 Canagliflozin也以不同剂量（3-30mg \u002F kg）给予ZDF大鼠4周，以评估其对BG对照和胰腺β细胞功能的影响。每周监测BG水平，并在4周治疗结束时测定HbA1c，血浆葡萄糖和胰岛素水平。在治疗4周后，在ZDF大鼠中进行口服葡萄糖耐量试验（OGTT）（2mg \u002F kg体重，通过管饲法给予）。在从尾静脉葡萄糖激发后0,30,60和120分钟对血液取样，使用血糖仪和血浆胰岛素使用ELISA方法测量BG水平。",{"key":251,"value":252},"参考文献","[1].\nLiang Y, et al. Effect of canagliflozin on renal threshold for glucose, glycemia, and body weight in normal and diabetic animal models. PLoS One. 2012;7(2):e30555.\n[2].\nKobuchi S, et al. A validated LC-MS\u002FMS method for the determination of canagliflozin, a sodium-glucose co-transporter 2 (SGLT-2) inhibitor, in a lower volume of rat plasma: application to pharmacokinetic studies in rats. 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