A novel breath test for the non-invasive assessment of small intestinal mucosal injury following methotrexate administration in the rat
A novel breath test for the non-invasive assessment of small intestinal mucosal injury following methotrexate administration in the rat
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DOI：
10.1080/00365520410003416
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发表时间：
2004-10-01
影响因子：
1.9
通讯作者：
Butler, RN
中科院分区：
文献类型：
--
作者：
Pelton, NS;Tivey, DR;Butler, RN
方法雄性Sprague道利大鼠（n= 8）每日3次皮下注射甲氨蝶呤（MTX：2.5 mg/kg）（第1-3天）以诱导小肠损伤（5）。对照组（n= 8）接受生理盐水注射。SBT在第7天进行，即动物处死前，收集小肠组织进行损伤的组织学评分和蔗糖酶活性的生化测定（5）。对于蔗糖呼吸试验，将大鼠禁食过夜，用2 mL蔗糖溶液（AnalaR，BDH，Merck Pty Ltd.，维多利亚，澳大利亚），并置于1 L有机玻璃容器中。当安装在灭菌舱上的双向旋塞关闭2 min时，向灭菌舱供应医用空气8 min，使13 CO2积聚。然后用连接到腔室出口的注射器收集呼吸样本（20 mL）2 min。每20 min重复该程序，共4 h。呼吸样本中的13 C水平用同位素比质谱仪测定。
MethodsMale Sprague Dawley rats (n= 8) received three daily subcutaneous methotrexate (MTX: 2.5 mg/kg) injections (days 1–3) to induce small intestinal damage (5). Controls (n= 8) received saline injections. The SBT was conducted on day 7, immediately before the animals were killed, when small intestinal tissues were collected for histological scoring of damage and biochemical determinations of sucrase activity (5). For the sucrose breath test, rats were fasted overnight, gavaged with a 2 mL sucrose solution (AnalaR, BDH, Merck Pty Ltd., Victoria, Australia) at 1 g/mL in water and placed in a 1L perspex container. Medical air was supplied to the chamber for 8 min when two-way taps fitted to the chamber were closed for 2 min, allowing the accumulation of 13CO2. Breath samples (20 mL) were then collected with a syringe attached to the chamber outlet for 2 min. This procedure was repeated every 20 min for a total of 4 h. 13C levels in breath samples were determined by isotope ratio mass spectrometer.