Fan Jun, Meng Qingyan, Guo Guanghua, Xie Yong, Li Xuedong, Xiu Yiping, Li Tairan, Feng Wei, Ma Liang
Department of Burns, The Northern Hospital, Shenyang 110015, Liaoning, PR China.
Burns. 2010 May;36(3):409-17. doi: 10.1016/j.burns.2009.05.020. Epub 2009 Sep 26.
This study aimed to investigate the influence of enteral glutamine (GLN) supplementation on Peyer's patch apoptosis in severely burned mice.
Thirty-four mice were randomly assigned to a normal group (n=10), an EN group (n=12) and an EN supplemented with GLN (EN+GLN) group (n=12) and the mice in the EN and EN+GLN groups received a full-thickness scald burn over 20% total body surface area (TBSA) on the back. The burned mice then were fed orally with a common EN or an isonitrogenous and isocaloric EN supplemented with GLN for 7 days. On day 7 after injury, all surviving mice were euthanised and the entire intestine was collected. The percentage of apoptotic cells and cell percentage of phenotype in Peyer's patches were determined by flow cytometry (FCM). The FasL expression in Peyer's patches was analysed by reverse transcription polymer chain reaction (RT-PCR) and FCM. Both TNF-alpha levels and caspase-3 activity in Peyer's patches were also assessed.
The results revealed that the percentage of lymphocyte subsets in Peyer's patches after burn injury significantly altered: the percentage of CD4 and CD19 cells declined and the percentage of CD8 cells correspondingly increased, when compared with the normal control mice (p<0.05). On the other hand, the total apoptotic ratio and all lymphocytes subset apoptosis in Peyer's patches were markedly increased (p<0.05), which were consistent with up-regulation in the FasL expression at the levels of both mRNA and protein, TNF-alpha levels and caspase-3 activity in Peyer's patches. Enteral GLN supplementation partially reversed these changes: the total apoptotic ratio and all lymphocytes subpopulation apoptosis in Peyer's patches were markedly decreased when compared with the EN group (p<0.05). The percentage of lymphocyte subsets within Peyer's patches also restored the condition prior to injury. However, no significant differences in the FasL expression, including mRNA and protein, were observed between the EN and EN+GLN groups. Although, both TNF-alpha levels and caspase-3 activity in Peyer's patches were lower in the EN+GLN group than in the EN group (p<0.05).
This study suggests that enteral GLN supplementation is superior to a common enteral nutrition with respect to attenuating apoptosis in Peyer's patches, which might be more effective in decreasing TNF-alpha levels and down-regulating caspase-3 activity in Peyer's patches.
本研究旨在探讨肠内补充谷氨酰胺(GLN)对严重烧伤小鼠派尔集合淋巴结细胞凋亡的影响。
34只小鼠随机分为正常组(n = 10)、肠内营养组(EN组,n = 12)和肠内补充谷氨酰胺组(EN + GLN组,n = 12)。EN组和EN + GLN组小鼠背部进行20%总体表面积(TBSA)的全层烫伤。随后,对烫伤小鼠经口给予普通肠内营养或补充谷氨酰胺的等氮等热量肠内营养,持续7天。在受伤后第7天,将所有存活小鼠安乐死并收集整个肠道。通过流式细胞术(FCM)测定派尔集合淋巴结中凋亡细胞的百分比和表型细胞百分比。通过逆转录聚合酶链反应(RT-PCR)和FCM分析派尔集合淋巴结中FasL的表达。同时评估派尔集合淋巴结中TNF-α水平和caspase-3活性。
结果显示,与正常对照小鼠相比,烧伤后派尔集合淋巴结中淋巴细胞亚群百分比发生显著变化:CD4和CD19细胞百分比下降,CD8细胞百分比相应增加(p < 0.05)。另一方面,派尔集合淋巴结中的总凋亡率和所有淋巴细胞亚群凋亡均显著增加(p < 0.05),这与派尔集合淋巴结中mRNA和蛋白水平的FasL表达上调、TNF-α水平和caspase-3活性一致。肠内补充GLN部分逆转了这些变化:与EN组相比,派尔集合淋巴结中的总凋亡率和所有淋巴细胞亚群凋亡均显著降低(p < 0.05)。派尔集合淋巴结内淋巴细胞亚群百分比也恢复到受伤前状态。然而,EN组和EN + GLN组之间在FasL表达(包括mRNA和蛋白)方面未观察到显著差异。尽管如此,EN + GLN组派尔集合淋巴结中的TNF-α水平和caspase-3活性均低于EN组(p < 0.05)。
本研究表明,在减轻派尔集合淋巴结细胞凋亡方面,肠内补充GLN优于普通肠内营养,可能在降低派尔集合淋巴结中TNF-α水平和下调caspase-3活性方面更有效。