Background
EIPA inhibited TRPP3-mediated Ca2+-activated currents with IC50 values of 10.5 μM [1]. TRPP3, a member of the transient receptor potential (TRP) superfamily of cation channels, is a Ca2+-activated channel permeable to Ca2+, Na+, and K+ [1].EIPA (5-(N-ethyl-N-isopropyl)-amiloride) is a commonly used amiloride derivative modified similarly to MPA, and its Ki for NHE1, NHE2, and NHE3 are 0.3, 1.8, and 67 μM, respectively[2,3]
EIPA suppressed proliferation of MKN28 cells through up-regulation of p21 expression via reduction of [Cl- ] c as a result from DIDS-sensitive Cl- /HCO3- exchanger-mediated compensation for keeping pHc normal under an NHE-inhibited condition [4].
EIPA preischemic treatment overcame the I/R-induced renal dysfunction. Histologic examination of the kidney of vehicle-treated ARF mice revealed severe lesions, and these lesions were significantly suppressed by the preischemic treatment with EIPA. In addition, EIPA suppressed the increment of renal ET-1 content after reperfusion [5]. EIPA at doses of 3 and 9 mg/kg per day inhibited tumor growth of HepG2 modle mice significantly compared with the control [6].
参考文献:
[1]. Dai XQ, Ramji A, Liu Y, Li Q, Karpinski E, Chen XZ (2007) Inhibition of TRPP3 channel by amiloride and analogs. Mol Pharmacol 72:1576-1585
[2]. Guffey, S.C.; Fliegel, L.; Goss, G.G. Cloning and characterization Na+/H+ Exchanger isoforms, NHE2 and, NHE3 from the gill Pacific dogfish, Squalus suckleyi. Comp. Biochem. Physiol. Part B Biochem. Mol. Biol. 2015, 188, 46-53.
[3]. Masereel, B. An overview inhibitors Na+/H+ exchanger. Eur. J. Med. Chem. 2003, 38, 547-554.
[4]. Hosogi SMH, Nakajima K, Ashihara E, Niisato N, Kusuzaki K, Marunaka Y: An inhibitor of Na(+)/H(+) exchanger (NHE), ethyl-isopropyl amiloride (EIPA), diminishes proliferation of MKN28 human gastric cancer cells by decreasing the cytosolic Cl(-) concentration via DIDS-sensitive pathways. Cell Physiol Biochem. 2012, 30: 1241-1253. 10.1159/000343315.
[5]. Yamashita J, Ohkita M, Takaoka M, Kaneshiro Y, Matsuo T,Kaneko K, Matsumura Y.Role of Na+/H+ exchanger in the pathogen-esis of ischemic acute renal failure in mice.J Cardiovasc Pharmacol49:154 -160, 2007.
[6]. X. Yang, D. Wang, W. Dong, Z. Song, K. Dou.Expression and modulation of Na+/H+ exchanger 1 gene in hepatocellular carcinoma: a potential therapeutic target. J. Gastroenterol. Hepatol., 26 (2) (2011), pp. 364-370
EIPA 抑制 TRPP3 介导的 Ca2+- 激活电流，IC50 值为 10.5 μM [1]。 TRPP3 是阳离子通道瞬时受体电位 (TRP) 超家族的成员，是 Ca2+- 激活通道，可渗透 Ca2+、Na+ 和 K+ [1].EIPA (5-(N-ethyl-N -isopropyl)-amiloride)是一种常用的类似MPA修饰的阿米洛利衍生物，其对NHE1、NHE2和NHE3的Ki值分别为0.3、1.8和67 μM[2,3] /p>\n
EIPA 通过减少 [Cl-] c 上调 p21 表达来抑制 MKN28 细胞的增殖，这是由于 DIDS 敏感的 Cl- /HCO3- 交换剂介导的补偿，以在 NHE 抑制条件下保持 pHc 正常[4].
EIPA 缺血前治疗克服了 I/R 引起的肾功能障碍。载体治疗的 ARF 小鼠肾脏的组织学检查显示严重病变，并且这些病变被 EIPA 的缺血前治疗显着抑制。此外，EIPA抑制再灌注后肾脏ET-1含量的增加[5]。与对照组相比，每天 3 和 9 mg/kg 剂量的 EIPA 显着抑制 HepG2 模型小鼠的肿瘤生长[6]。
Protocol
| Cell experiment [1]: |
Cell lines | MCF-7 cells |
Preparation Method | Cells were seeded in 6-well plates for 24 h prior to 48 h-treatment with EIPA (10 μM), Akti (1 μM), U0126 (10 μM) or a combination thereof. |
Reaction Conditions | 10μM for 48 hours |
Applications | MCF-7 cells were subjected to treatment with EIPA with and without ERK and Akt inhibitors. ERK, but not Akt, activity was increased upon EIPA treatment, and both kinases were effectively inhibited by their inhibitors, U0126 and Akti, respectively. Strikingly, ERK inhibition strongly potentiated EIPA-induced CHOP upregulation and PARP cleavage. |
| Animal experiment [2]: |
Animal models | Female homozygous BALB/c Nu/Nu nude mice were injected subcutaneously (s.c.) in both flanks at 7 weeks of age with 106 parental and IKKαδ MIA PaCa-2 cells or 1334 and 1444 cells mixed at a 1:1 dilution with BD Matrigel (BD Biosciences) in a total volume of 100 μl. |
Preparation Method | Mice were treated with vehicle (DMSO in PBS), EIPA (7.5 mg/kg), MRT68921 (10 mg/kg) or MRT68921+EIPA for 15 days when tumors attained an average volume of 50-100 mm3. |
Dosage form | s.c., 7.5 mg/kg/d for 7 days |
Applications | One month of EIPA treatment, initiated at age 1 month, markedly reduced pancreatic weight (a tumor burden surrogate), inhibited acinar-to-ductal metaplasia (ADM) and advanced PanIN formation, and preserved the normal pancreatic parenchyma in KrasG12D;IkkαδPEC mice, while decreasing MP. |
参考文献: [1]: M.G. Rolver, L.O. Elingaard-Larsen, A.P. Andersen, L. Counillon, S.F. Pedersen. Pyrazine ring-based Na+/H+ exchanger (NHE) inhibitors potently inhibit cancer cell growth in 3D culture, independent of NHE1.Sci. Rep., 10 (2020), p. 5800
[2]: H. Su, F. Yang, R. Fu, et al. Cancer cells escape autophagy inhibition via NRF2-induced micropinocytosis. Cancer Cell, 39 (5) (2021), pp. 678-693, 10.1016/j.ccell.2021.02.016 |
没有评价数据