Braga-Neto Manuel B, Warren Cirle A, Oriá Reinaldo B, Monteiro Manuel S, Maciel Andressa A S, Brito Gerly A C, Lima Aldo A M, Guerrant Richard L
Institute of Biomedicine and Clinical Research Unit-University Hospital, Federal University of Ceará, Fortaleza, Brazil.
Dig Dis Sci. 2008 Oct;53(10):2687-96. doi: 10.1007/s10620-008-0215-0. Epub 2008 Mar 6.
In this study, we have examined the role of glutamine derivatives in reducing 5-fluorouracil (5-FU)-induced epithelial damage in an undifferentiated crypt intestinal cell line, IEC-6. In this model, we have investigated proliferation indirectly by detecting the enzyme-derived formazan dye from the tetrazolium salt WST-1 in viable cells at 24 and 48 h after 5-FU treatment. Migration was measured at 12 and 24 h after razor scraping of the cell monolayer. Cell death was measured by quantifying the percentage of apoptotic and necrotic figures by flow cytometry at 12 and 24 h following 5-FU challenge. Neither glutamine nor alanyl-glutamine prevented 5-FU-induced apoptosis and necrosis in IEC-6 cells at 12 and 24 h after 5-FU challenge. However, glutamine and alanyl-glutamine enhanced migration and proliferation when compared with 5-FU-treated controls (P < 0.05). These new findings support our earlier study on the benefit of oral glutamine in enhancing epithelial recovery after 5-FU challenge.
在本研究中,我们检测了谷氨酰胺衍生物在减轻5-氟尿嘧啶(5-FU)诱导的未分化隐窝肠上皮细胞系IEC-6上皮损伤中的作用。在该模型中,我们通过在5-FU处理后24小时和48小时检测活细胞中四唑盐WST-1的酶促生成的甲臜染料来间接研究细胞增殖。在刮伤细胞单层后12小时和24小时测量细胞迁移。在5-FU刺激后12小时和24小时,通过流式细胞术对凋亡和坏死细胞的百分比进行定量来测量细胞死亡。在5-FU刺激后12小时和24小时,谷氨酰胺和丙氨酰谷氨酰胺均不能预防5-FU诱导的IEC-6细胞凋亡和坏死。然而,与5-FU处理的对照组相比,谷氨酰胺和丙氨酰谷氨酰胺可增强细胞迁移和增殖(P<0.05)。这些新发现支持了我们早期关于口服谷氨酰胺对5-FU刺激后上皮恢复有益的研究。