CAJ | 학술논문

目的:研究丙氨酰-谷氨酰胺(Ala-Gln)对口服FK506损伤的肠黏膜组织中iNOS和TNF-α分子表达的影响.方法:BALB/c小鼠24只,随机分成对照组、FK506低剂量组、FK506高剂量组及Ala-Gln治疗组,分别给以0.2 mL生理盐水、FK506 0.1 mg/kg、1.0 mg/kg灌胃和FK506 1.0 mg/kg灌胃及丙氨酰-谷氨酰胺0.5 g/kg腹腔注射.隔天给药,6周后采集回肠标本.HE染色和扫描电镜观察肠黏膜组织形态学改变;FITC-dextran(FD4)检测肠黏膜通透性;RT-PCR检测小肠黏膜iNOS和TNF-α mRNA的表达情况;Western blotting检测iNOS和TNF-α蛋白表达水平.结果:FK506高剂量组的肠黏膜对FD4的通透性明显增加,扫描电镜示小肠绒毛破坏明显,而Ala-Gln治疗组小肠绒毛破坏减轻,对FD4的通透性下降;FK506高剂量组小肠黏膜iNOS mRNA和TNF-α mRNA表达增强,而Ala-Gln治疗组表达则明显下调;iNOS和TNF-α蛋白表达水平的变化与此一致.结论:FK506通过上调iNOS和TNF-α的表达对小肠黏膜产生损伤,使小肠壁的通透性增加.Ala-Gln对FK506所致的肠黏膜屏障功能损伤具有保护作用,该作用可能与下调iNOS和TNF-α的表达有关.
목적:연구병안선-곡안선알(Ala-Gln)대구복FK506손상적장점막조직중iNOS화TNF-α분자표체적영향.방법:BALB/c소서24지,수궤분성대조조、FK506저제량조、FK506고제량조급Ala-Gln치료조,분별급이0.2 mL생리염수、FK506 0.1 mg/kg、1.0 mg/kg관위화FK506 1.0 mg/kg관위급병안선-곡안선알0.5 g/kg복강주사.격천급약,6주후채집회장표본.HE염색화소묘전경관찰장점막조직형태학개변;FITC-dextran(FD4)검측장점막통투성;RT-PCR검측소장점막iNOS화TNF-α mRNA적표체정황;Western blotting검측iNOS화TNF-α단백표체수평.결과:FK506고제량조적장점막대FD4적통투성명현증가,소묘전경시소장융모파배명현,이Ala-Gln치료조소장융모파배감경,대FD4적통투성하강;FK506고제량조소장점막iNOS mRNA화TNF-α mRNA표체증강,이Ala-Gln치료조표체칙명현하조;iNOS화TNF-α단백표체수평적변화여차일치.결론:FK506통과상조iNOS화TNF-α적표체대소장점막산생손상,사소장벽적통투성증가.Ala-Gln대FK506소치적장점막병장공능손상구유보호작용,해작용가능여하조iNOS화TNF-α적표체유관.
AIM: To investigate the effects of alanyl - glutamine ( Ala -Gln) on expression of iNOS and TNF- α in injured intestinal mucosa induced by oral tacrolimus (FK506). METHODS: Twenty -four BALB/c mice were randomized to receive orally 0.2 mL of normal saline solution ( group Ⅰ ), 0.2 mL of FK506 in a dose of 0.1 mg/kg ( group Ⅱ ) or 1.0 mg/kg (group Ⅲ), and orally high -dose FK506 (0.2 mL, 1.0 mg/kg) plus intraperitoneal injection of Ala -Gln (0.5 g/kg )(group Ⅳ ),respectively. Damages of intestinal mucosa were determined by pathological examination.Intestinal mucosal permeability was analysed by FITC - dextran fluorescence assay. Expression of iNOS and TNF - α in intestine was detected by RT - PCR and Western blotting. RESULTS: Severe damage on the villi and increased intestinal permeability were observed in high - dose FK506 treated mice according to scanning electron microscopy and FITC - dextran flux respectively. The erosion and increased intestinal permeability were significantly alleviated by Ala - Gln treatment. Transcription of iNOS mRNA and TNF - α mRNA, which was up - regulated in high - dose FK506 treated group,was also markedly down- regulated in mice combined with Ala- Gln- treatment. A significantly increased expression of iNOS and TNF - α protein was found in the high - dose FK506 treated mice, while small amounts of these proteins were identified in the Ala - Gln - treated group. CONCLUSION: FK506 could induce a significant impairment of intestinal mucosa morphologically, which might be associated with up - regulated expression of iNOS and TNF - α in small intestinal mucosa. Subsequently, the intestinal permeability is increased. Ala - Gln has a strong protective effect on FK506 - induced intestinal barrier dysfunction, probably relates to the down - regulation of iNOS and TNF - α expression.