Alanyl-glutamine attenuates 5-fluorouracil-induced intestinal mucositis in apolipoprotein E-deficient mice.
Alanyl-glutamine attenuates 5-fluorouracil-induced intestinal mucositis in apolipoprotein E-deficient mice.
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DOI：
10.1590/1414-431x20144360
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发表时间：
2015-06
期刊：
影响因子：
--
通讯作者：
Oriá RB
中科院分区：
文献类型：
--
作者：
Araújo CV;Lazzarotto CR;Aquino CC;Figueiredo IL;Costa TB;Alves LA;Ribeiro RA;Bertolini LR;Lima AA;Brito GA;Oriá RB
载脂蛋白E（APOE=基因，apoE=蛋白）是一种已知的调节炎症反应的因子，在组织从损伤中恢复期间可能具有再生作用。我们研究了在5-FU诱导的肠粘膜炎后的组织恢复过程中，apoE缺乏是否会降低丙氨酰谷氨酰胺（Ala-Gln）治疗（一种公认的肠道营养素）的愈合效果。在5-氟尿嘧啶（5-FU）激发（450 mg/kg，通过腹膜内注射）前3天和后5天，通过管饲法给予APOE敲除（APOE-/-）和野生型（APOE+/+）C57 BL 6 J雄性和雌性小鼠（N=86）Ala-Gln（100 mM）或磷酸盐缓冲盐水（PBS）。每天监测小鼠体重。白细胞计数法评价5-FU的细胞毒作用。监测肠绒毛高度、绒毛/隐窝比和绒毛蛋白表达以评估肠吸收表面积的恢复。使用隐窝长度、有丝分裂、凋亡和坏死隐窝指数以及胰岛素样生长因子-1（IGF-1）和B细胞淋巴瘤2（Bcl-2）肠道mRNA转录本的定量实时PCR来评估肠道上皮细胞更新。5-FU攻击导致两种小鼠品系的体重减轻和白细胞减少（P<0.001），Ala-Gln没有改善。所有5-FU攻击小鼠均出现绒毛钝化、隐窝增生和绒毛/隐窝比降低（P<0.05），但PBS对照组未出现。与PBS对照组相比，Ala-Gln改善了所有激发小鼠的绒毛/隐窝比、隐窝长度和有丝分裂指数。Ala-Gln仅在APOE-/-小鼠中改善绒毛高度。与未处理的对照组相比，5-FU激发的所有小鼠的隐窝细胞凋亡和坏死评分均增加。Ala-Gln处理的APOE+/+小鼠的这些评分显著低于对照组。Bcl-2和IGF-1 mRNA转录仅在APOE-/--攻击的小鼠中减少。总之，我们的研究结果表明，5-FU激发后，APOE独立的Ala-Gln再生效应。
Apolipoprotein E (APOE=gene, apoE=protein) is a known factor regulating the inflammatory response that may have regenerative effects during tissue recovery from injury. We investigated whether apoE deficiency reduces the healing effect of alanyl-glutamine (Ala-Gln) treatment, a recognized gut-trophic nutrient, during tissue recovery after 5-FU-induced intestinal mucositis. APOE-knockout (APOE-/-) and wild-type (APOE+/+) C57BL6J male and female mice (N=86) were given either Ala-Gln (100 mM) or phosphate buffered saline (PBS) by gavage 3 days before and 5 days after a 5-fluorouracil (5-FU) challenge (450 mg/kg, via intraperitoneal injection). Mouse body weight was monitored daily. The 5-FU cytotoxic effect was evaluated by leukometry. Intestinal villus height, villus/crypt ratio, and villin expression were monitored to assess recovery of the intestinal absorptive surface area. Crypt length, mitotic, apoptotic, and necrotic crypt indexes, and quantitative real-time PCR for insulin-like growth factor-1 (IGF-1) and B-cell lymphoma 2 (Bcl-2) intestinal mRNA transcripts were used to evaluate intestinal epithelial cell turnover. 5-FU challenge caused significant weight loss and leukopenia (P<0.001) in both mouse strains, which was not improved by Ala-Gln. Villus blunting, crypt hyperplasia, and reduced villus/crypt ratio (P<0.05) were found in all 5-FU-challenged mice but not in PBS controls. Ala-Gln improved villus/crypt ratio, crypt length and mitotic index in all challenged mice, compared with PBS controls. Ala-Gln improved villus height only in APOE-/- mice. Crypt cell apoptosis and necrotic scores were increased in all mice challenged by 5-FU, compared with untreated controls. Those scores were significantly lower in Ala-Gln-treated APOE+/+ mice than in controls. Bcl-2 and IGF-1 mRNA transcripts were reduced only in the APOE-/--challenged mice. Altogether our findings suggest APOE-independent Ala-Gln regenerative effects after 5-FU challenge.