Empagliflozin (BI 10773) is an SGLT-2 inhibitor (IC50=3.1 nM) that is potent and selective, with more than 300-fold selectivity for SGLT-1/4/5/6. Empagliflozin is used for the treatment of type 2 diabetes.

SGLT2:3.1 nM

方法：猪肾细胞 LLC-PK1 用 Empagliflozin (100-500 nM) 和高糖 HG (30 mM) 处理 24 h，使用 Glutathione Assay Kit 检测 GSH 水平。
结果：在用 HG30 处理的细胞中，GSH 含量相对于对照降低。在用 HG30 处理的细胞中添加 Empagliflozin 后，GSH 水平提高。[1]
方法：人 PTC 细胞 HK-2 用 Empagliflozin (100-500 nM) 和 glucose (30 mM) 处理 24 h，使用 TUNEL assay 检测细胞凋亡情况。
结果：高糖显著诱导细胞凋亡，Empagliflozin 可以改善这种作用。[2]

方法：为研究对糖尿病小鼠心肌损伤的影响，将 Empagliflozin (10 mg/kg in 0.5% hydroxyethylcellulose) 灌胃给药给糖尿病 KK-Ay 小鼠，每天一次，持续八周。
结果：Empagliflozin 改善了糖尿病心肌的结构和功能，降低了心肌氧化应激，改善了心肌纤维化。Empagliflozin 通过抑制 TGFβ/Smad 通路和激活 Nrf2/ARE 信号传导来抑制氧化应激和纤维化。[3]
方法：为研究对 1 型糖尿病的作用，将 Empagliflozin (3-10 mg/kg in hydroxy ethyl cellulose) 灌胃给药给 streptozotocin 诱导 T1DM 的 C57BL/6J 小鼠，每天一次，持续八天。
结果：Empagliflozin 治疗使 1 型糖尿病小鼠的糖耐量显著改善，胰岛素 mRNA 表达增加，血清胰岛素水平增加。Empagliflozin 可能通过保护胰腺 β 细胞免受糖毒性诱导的氧化应激，对保护 β 细胞再生，从而改善 1 型糖尿病患者的血糖稳态具有有益作用。[4]

[14C]-monosaccharide uptake inhibition experiments: Stable cell lines over-expressing hSGLT-1, -2, -4, -5 or -6 or rSGLT-1 or -2 are used for the sodium-dependent monosaccharide transport inhibition assay. Cells are pre-incubated in 200 μL uptake buffer (10 mM HEPES, 137 mM NaCl, 5.4 mM KCl, 2.8 mM CaCl2, 1.2 mM MgCl2, 50 μg/ml Gentamycin, 0.1% BSA) for 25 minutes at 37°C. 10 μM Cytochalasin B and test compound is added at different concentrations 15 minutes before the initiation of the uptake experiment. The uptake reaction is started by the addition of 0.6 μCi [14C]-labelled monosaccharide i.e. [14C]-labelled AMG, glucose, fructose, mannose or myo-inositol, in 0.1 mM AMG (or the respective non-radioactive monosaccharide). After incubation for 60 minutes (hSGLT-5), 90 minutes (hSGLT-4) or 4 hours (hSGLT-2) at 37°C, the cells are washed three times with 300 μL PBS and then lysed in 0.1 N NaOH with intermittent shaking for 5 minutes. The lysate is mixed with 200 μL MicroScint 40 and shaken for 15 minutes and counted for radioactivity in the TopCount NXT. For SGLT-4 and SGLT-5 assays cells are pre-incubated in pre-treatment buffer (uptake buffer containing choline chloride instead of NaCl) for 25 minutes prior to addition of uptake buffer.

MTS assay(Only for Reference)