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谷氨酰胺增强术后患者中性粒细胞对细菌的杀伤作用。

Glutamine-enhanced bacterial killing by neutrophils from postoperative patients.

作者信息

Furukawa S, Saito H, Fukatsu K, Hashiguchi Y, Inaba T, Lin M T, Inoue T, Han I, Matsuda T, Muto T

机构信息

Department of Surgery, Faculty of Medicine, University of Tokyo, Japan.

出版信息

Nutrition. 1997 Oct;13(10):863-9. doi: 10.1016/s0899-9007(97)00271-2.

Abstract

Neutrophils play an important role in host defense by phagocytosing and destroying invading bacteria. A recent investigation revealed that glutamine (Gln) augmented the in vitro bactericidal activity of neutrophils from burn patients. However, it is unclear whether Gln enhances the function of neutrophils in postoperative patients. This study was designed to investigate the effect of Gln on the in vitro Escherichia coli-killing activity of neutrophils from postoperative patients. Nine randomly selected patients were included in this study. On the morning of the first postoperative day, blood was drawn and neutrophils were isolated. Eight healthy volunteers served as controls. E. coli was opsonized with pooled normal serum. Neutrophils (5 x 10(6)), together with opsonized E. coli (5 x 10(5)), were incubated for 2 h at 37 degrees C in Hanks' balanced salt solution supplemented with 0, 100, 500, or 1000 nmol/mL of Gln. The bactericidal function of neutrophils was determined by counting the number of viable bacteria. Tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 beta, IL-8, and granulocyte elastase levels in the cell culture supernatant were measured. Plasma C-reactive protein (CRP), cortisol, and amino acids were also analyzed. The plasma concentration of Gln was significantly lower in the postoperative patients than in the controls. Following culture with patient neutrophils, the number of viable E. coli decreased by 26% as the in vitro Gln concentration was increased from 500 to 1000 nmol/mL (P < 0.01). We defined the Gln 1000/Gln 500 ratio of the number of viable bacteria as the number of viable E. coli at an in vitro Gln concentration of 1000 nmol/mL divided by the number of viable E. coli at an in vitro Gln concentration of 500 nmol/mL. A positive correlation was thus demonstrated between the plasma Gln level and the Gln 1000/Gln 500 ratio of the number of viable bacteria in the patients (r = 0.69, P = 0.04). This finding indicated that as plasma Gln fell, there was an enhancement of neutrophil E. coli-killing activity by neutrophils in in vitro tests when the Gln concentration was increased from 500 to 1000 nmol/mL. Gln supplementation caused no appreciable changes in TNF-alpha, IL-1 beta, IL-8, or granulocyte elastase levels in cell culture supernatants. A negative correlation was recognized between the patient plasma Gln level and the Gln 1000/Gln 500 ratio of the cell culture supernatant IL-8 level (r = -0.73, P = 0.025). In conclusion, Gln supplementation enhanced the in vitro bactericidal function of neutrophils from postoperative patients.

摘要

中性粒细胞通过吞噬和破坏入侵细菌在宿主防御中发挥重要作用。最近的一项研究表明,谷氨酰胺(Gln)可增强烧伤患者中性粒细胞的体外杀菌活性。然而,尚不清楚Gln是否能增强术后患者中性粒细胞的功能。本研究旨在探讨Gln对术后患者中性粒细胞体外杀灭大肠杆菌活性的影响。本研究纳入了9名随机选择的患者。在术后第一天上午采血并分离中性粒细胞。8名健康志愿者作为对照。用混合正常血清调理大肠杆菌。将中性粒细胞(5×10⁶)与调理后的大肠杆菌(5×10⁵)在补充有0、100、500或1000 nmol/mL Gln的汉克斯平衡盐溶液中于37℃孵育2小时。通过计算活菌数量来确定中性粒细胞的杀菌功能。测量细胞培养上清液中的肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、IL-8和粒细胞弹性蛋白酶水平。还分析了血浆C反应蛋白(CRP)、皮质醇和氨基酸。术后患者的血浆Gln浓度显著低于对照组。在用患者中性粒细胞培养后,随着体外Gln浓度从500 nmol/mL增加到1000 nmol/mL,活菌大肠杆菌数量减少了26%(P<0.01)。我们将活菌数量的Gln 1000/Gln 500比值定义为体外Gln浓度为1000 nmol/mL时的活菌大肠杆菌数量除以体外Gln浓度为500 nmol/mL时的活菌大肠杆菌数量。因此,在患者中血浆Gln水平与活菌大肠杆菌数量的Gln 1000/Gln 500比值之间呈正相关(r = 0.69,P = 0.04)。这一发现表明,当血浆Gln下降时,在体外试验中,当Gln浓度从500 nmol/mL增加到1000 nmol/mL时,中性粒细胞杀灭大肠杆菌的活性增强。补充Gln对细胞培养上清液中的TNF-α、IL-1β、IL-8或粒细胞弹性蛋白酶水平没有明显影响。患者血浆Gln水平与细胞培养上清液IL-8水平的Gln 1000/Gln 500比值之间呈负相关(r = -0.73,P = 0.025)。总之,补充Gln可增强术后患者中性粒细胞的体外杀菌功能。

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