Toll样受体-2在早产大鼠坏死性小肠结肠炎中的表达及谷氨酰胺对其的影响  

Expression of Toll-like receptor-2 in premature rats with necrotizing enterocolitis and regulatory effects of glotamine

作　　者：李薇[1] 郑晓辉[1] 周伟 荣箫 黄龙光 

机构地区：[1]广东省东莞市太平人民医院儿科,523905 [2]广州市妇女儿童医疗中心广州市儿童医院新生儿科

出　　处：《中华围产医学杂志》2011年第5期294-299,共6页Chinese Journal of Perinatal Medicine

基　　金：广东省自然科学基金（8151012001000002）

摘　　要：目的 探讨坏死性小肠结肠炎（necrotizing enterocolitis，NEC）早产大鼠肠道组织中Toll样受体-2（Toll—likereceptor-2，TLR-2）和半胱氨酸蛋白酶～3（caspase-3）的表达及谷氨酰胺（glutamine，Gln）对其表达的影响及对NEC肠道损伤的保护作用。方法60只早产新生大鼠按随机数字表法分为模型组、Gln干预组和对照组，每组20只。模型组和Gln干预组早产新生大鼠采用代乳品人工喂养＋缺氧冷刺激建立NEC模型，Gin干预组同时在配方乳中加入Gin0．3g／kg；对照组未进行任何干预。所有早产大鼠生后第3天处死后取肠道组织，HE染色观察病理改变，对回肠进行病理评分，并采用免疫组织化学方法检测空肠、回肠和结肠caspase-3、TLR-2蛋白的表达，采用荧光定量聚合酶链反应技术检测空肠、回肠及结肠TLR-2mRNA的表达。结果模型组、Gln干预组和对照组回肠损伤病理评分分别为（3．10±0．99）分、（2．40±0．69）分及（0．30±0．48）分；回肠组织TLR-2蛋白的表达分别为2．53±0．94、2．15±0．82及1．57±0．62；caspase-3蛋白的表达分别为2．83土0．45、2．70±0．04及0．91±0．29；模型组TLR-2tuRNA的含量为Gin干预组的1．46倍，为对照组的2．10倍。与对照组相比，模型组的肠道损伤病理评分、caspase-3蛋白、TLR-2蛋白和TLR-2mRNA的表达均增加（P均〈0．01）；与模型组相比，Gln干预组的肠道损伤病理评分、caspase-3蛋白、TLR-2蛋白和TLR-2mRNA的表达均有下降（P〈0．05）。TLR-2mRNA和caspase-3蛋白的表达呈正相关（r=0．71，P〈0．01），与肠组织损伤病理评分也呈正相关（r=0．69，P〈0．01）；caspase-3与肠组织损伤病理评分呈正相关（r=0．81，P〈0．01）。结论TLR-2可能参与了NEC的发病。降低TLR-2的表达，抑制肠黏膜细胞的凋亡，可能是Gln保护NEC早产大鼠肠道的机制之一。Objective To study the expression of Toll-like receptor-2 （TLR-2） and caspase-3 in the intestine of premature rats with necrotizing enterocolitis （NEC）, and to explore the protective effects and possible regulatory mechanism of glutamine （Gln） in the NEC. Methods Sixty premature rats （gestational age 21d） were divided into three groups （n=20 each） according to the random number table： control group, model group and Gln intervention group. Rats in model group were given formula feeding, hypoxia and cold stress. Rats in Gln intervention group were given Gin 0.3 g/kg to the formula feeding, hypoxia and cold stress. All the premature rats were sacrificed and the intestine tissues were obtained on the third day after birth. The histological changes of ileal tissues were scored after HE staining. The expression of TLR-2 and caspase-3 in jejunum, ileum and colon were detected by inmunohistochemistry, and the expression of TLR-2 mRNA in jejunum, ileum and colon were detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction. Results Pathology score of ileum in model group, Gln intervention group and control group were 3.10 ± 0.99, 2.40±0.69 and 0.30± 0.48, respectively. The expressions of TLR-2 protein in ileum were 2.53±0.94, 2.15±0.82 and 1.57±0.62 in the three groups respectively, and the expression of easpase 3 protein were 2.83±0.45, 2.70±0.04 and 0.91±0.29. The content of TLR2 mRNA in model group was 1.46 times higher than that of Gln intervention group and was 2. 10 times higher than that of control group. Compared with the control group, the pathology score, expression of TLR 2 and caspase 3 protein, and TLR-2 mRNA in model group were significantly higher, P〈0.01. However, compared with the model group, those changes were improved in Gin intervention group, P〈0.05. Expression of TLR-2 mRNA positively correlated to the expression of easpase 3 protein （r=0.71, P〈0.01） and pathology score （r=0.69,P（0.01）. Expression of caspase-3 prot

关 键 词：小肠结肠炎 坏死性 谷氨酰胺 TOLL样受体2 细胞凋亡 早产 大鼠 

分 类 号：R714.2[医药卫生—妇产科学]