谷氨酰胺对内毒素血症新生幼鼠肠道功能保护作用的研究  

The effect of glutamine on gastrointestinal protection in lipopolysaccharide-induced newborn rats

作　　者：苗士建[1] 张烨[1] 黄瑛[1] 陈莲[2] 

机构地区：[1]复旦大学附属儿科医院消化科,上海201102 [2]复旦大学附属儿科医院病理科,上海201102

出　　处：《中国循证儿科杂志》2012年第6期459-464,共6页Chinese Journal of Evidence Based Pediatrics

基　　金：上海市自然科学基金:062R14017;卫生部国家临床重点专科建设项目

摘　　要：目的观察谷氨酰胺(Gln)对内毒素血症新生幼鼠模型肠黏膜的保护作用。方法 4窝清洁级SD母鼠及其新生幼鼠随机分为蛋氨酸亚砜胺(MS)+生理盐水(NS)组(母鼠和幼鼠给予MS+幼鼠给予NS)、MS+Gln组(母鼠和幼鼠给予MS+幼鼠给予Gln)、正常喂养+Gln组(幼鼠正常母乳喂养+幼鼠给予Gln)和正常喂养+NS组(幼鼠正常母乳喂养+幼鼠给予NS)。各组干预均从母鼠分娩第3天和幼鼠出生第3天起开始,连续给予6d。实验第7天,各组幼鼠均腹腔注射LPS,6h后处死,测定血浆Gln浓度,肠上皮细胞膜脂质流动性及IL-1、TNF-α、MCP-1的水平,光镜、扫描电镜和透射电镜下观察肠黏膜绒毛结构的改变。结果①MS+Gln组、MS+NS组、正常喂养+Gln组和正常喂养+NS组出生幼鼠分别为11、11、10和11只,MS+Gln组和MS+NS组在实验结束前分别死亡2(18.2%)和6只(54.5%)。②MS+Gln组、MS+NS组、正常喂养+Gln组和正常喂养+NS组:体重增加量分别为(1.52±0.64)、(0.58±0.77)、(11.72±1.03)和(11.80±1.45)g,血浆Gln浓度分别为(394.3±92.5)、(105.9±37.9)、(774.6±143.7)和(676.8±113.6)μmol·L-1,肠上皮细胞膜荧光偏振度分别为(0.241±0.046)、(0.297±0.033)、(0.215±0.052)和(0.211±0.049),MCP-1水平分别为(8.0±1.6)、(5.8±0.8)、(8.5±1.3)和(8.7±1.2)pg·mL-1,TNF-α水平分别为(2.2±0.8)、(1.3±0.4)、(2.8±1.0)和(2.5±1.1)pg·mL-1,IL-1β水平分别为(28.5±15.5)、(16.8±11.0)、(31.4±15.1)、(32.3±13.9)pg·mL-1;组间总体比较差异均有统计学意义(均P<0.05)。③光镜下可见:MS+NS组和MS+Gln组肠黏膜绒毛均有不同程度变短,排列杂乱,黏膜结构破坏,以MS+NS组程度更重;正常喂养+Gln组和正常喂养+NS组肠黏膜绒毛亦有不同程度的排列紊乱和变短,但较MS+Gln组损伤程度轻。④电镜下可见:MS+NS组和MS+Gln组均有肠黏膜微绒毛排列杂乱,长度缩短,断裂明显,肠上皮细胞绒毛膜欠完整,以MS+NS组程度更重;正常喂养+Gln组及正常喂养+NS组肠黏�Objective To study the protective effects of glutamine（Gln） on intestinal mucosa in lipopolysaccharide-induced newborn rats.Methods Four broods of mother and infant rats were randomized into four groups： methionine sulfoximine（MS）＋Gln group： Gln-deprived pups ＋ Gln supplementation ;MS＋NS group： Gln-deprived pups without Gln supplementation; Normally fed and Gln group： normally fed pups ＋ Gln supplementation; Normally fed and NS group： normally fed pups without Gln supplementation; MS,NS and Gln were given by intraperitoneal injection for 6 days. All pups received lipopolysaccharide （LPS） on D7 and were sacrificed 6 hours later. Small intestine was harvested for studies. Plasma Gln concentration was detected by high efficiency liquid chromatography. IL-1, TNF-α and MCP-1 levels were determined with homogenate by ELISA. The intestines were examined under both light, transmission and scanning electron microscopy （EM） and membrane fluidity was determined with fluorescent polarization, using a DPH probe.Results ①The number of pups in 4 groups was 11, 11, 10 and 11 respectively, but when the experiment was finished,the number was 9, 5, 10 and 11 respectively. ②Pups weight gain （g）, plasma Gln concentration（μmol·L-1）, IEC membrane fluorescence polarization and cytokine expression （MCP-1, TNF-α, IL-1β pg·mL-1）of four groups were（1.52±0.64）,（0.58±0.77）,（11.72±1.03）and （11.80±1.45）,（394.3±92.5）,（105.9±37.9）,（774.6±143.7）and （676.8±113.6）,（0.241±0.046）,（0.297±0.033）,（0.215±0.052）and （0.211±0.049）,（8.0±1.6）、（5.8±0.8）、（8.5±1.3）and（8.7±1.2）,（2.2±0.8）,（1.3±0.4）,（2.8±1.0）and （2.5±1.1）,（28.5±15.5）,（16.8±11.0）,（31.4±15.1）and（32.3±13.9）. Overall, the between-group differences were statistically significant（P0.05）.③Under light microscopy,the intestinal structure of MS＋NS group pups was not intact, small intestine villi were the shortest and badly d

关 键 词：谷氨酰胺 蛋氨酸亚砜胺 脂多糖 细胞膜脂质流动性 

分 类 号：R722.1[医药卫生—儿科]