目的 测定急性脊髓损伤大鼠血清中二胺氧化酶(DAO)的含量,同时观察肠道运动功能的变化和肠黏膜屏障损伤的程度。方法 45只 Sprague-Dawley大鼠随机分为打击组(A组)、假手术组(B组)和对照组(C组),每组15只。采取改良Allen打击法(10 g×25 mm)损伤大鼠T10脊髓节段。术后1 d、3 d、7 d分别对各组大鼠进行后肢功能BBB评分,测定大鼠回肠肌电慢波及平滑肌收缩,并做回肠HE染色,ELISA试剂盒测定血清DAO含量。结果 脊髓损伤后1 d、3 d和7 d,A组BBB评分显著低于C组(P<0.001),B组与C组无显著性差异(P>0.05)。脊髓损伤后1 d、3 d,A组慢波频率和振幅低于C组(P<0.05),收缩频率和振幅低于C组(P<0.05),B组与C组无显著性差异(P>0.05);脊髓损伤后7 d,三组肌电慢波频率和振幅、收缩的频率和振幅均无显著性差异(P>0.05)。A组回肠黏膜水肿、倒伏,炎性细胞浸润,黏膜下间隙增大。A组肠黏膜损伤Chiu评分高于B组和C组(P<0.05)。脊髓损伤后1 d、3 d,A组大鼠血清DAO含量高于C组(P<0.05),B组与C组无显著性差异(P>0.05);脊髓损伤后7 d,三组间血清DAO含量均无显著性差异(P>0.05)。结论 血清DAO含量可以反映大鼠急性脊髓损伤后肠道的运动功能和黏膜损伤程度。
Objective To measure the level of diamine oxidase (DAO), and observe the intestinal motor and mucosal barrier injury after acute spinal cord injury (SCI) in rats. Methods A total of 45 Sprague-Dawley rats were randomly divided into SCI group (group A, n=15), sham group (group B, n=15) and control group (group C, n=15). SCI model was established with Allen's strike mode (10 g × 25 mm) by striking T10 spinal segment in rats. One day, three days and seven days after SCI, hind limb motor function was assessed with Basso-Beattie-Bresnahan (BBB) Scale in each group, the myoelectric slow wave and ileum smooth muscle contractility were measured in rats, ileum tissues were tested with HE staining, and the DAO content of serum was tested with ELISA kit. Results One day, three days and seven days after SCI, the BBB score was significantly lower in group A than in groups B and C (P<0.001). One day, three days after SCI, the frequency and amplitude of both slow wave and contractility were lower in group A than in groups B and C (P<0.05); seven days after SCI, there was no significant difference among three groups (P>0.05). Group A showed ileal mucosal edema, lodging, inflammatory cell infiltration, and submucosal gap increase. The Chiu's score of intestinal mucosal injury was higher in group A than in groups B and C (P<0.05), as well as the serum DAO content one day and three days after SCI (P<0.05), and no significant difference was found in the serum DAO content among three groups seven days after SCI (P>0.05). Conclusion Serum DAO content may respond to the intestinal motor function and mucosal injury after acute SCI in rats.
[1] 刘金辉,易满. 肠道功能训练用于脊髓损伤患者的临床效果评价[J]. 世界最新医学信息文摘, 2015, 15(101): 102-104.
[2] 崔勇,段佩琰,刘劲松. 肠道训练联合饮食指导对脊髓损伤患者肠道康复效果的影响[J]. 中国现代药物应用, 2016, 10(1): 212-213.
[3] 陈红霞. 脊髓损伤患者肠道功能重建[J]. 吉林医学, 2013, 34(17): 3460-3461.
[4] 朱黎婷,朱毅,张文毅,等. 中医药在脊髓损伤神经源性肠道功能障碍的研究进展[J]. 世界华人消化杂志, 2012, 20(35): 3549-3557.
[5] 徐青,高飞,王磊,等. 脊髓损伤后肠道功能障碍：美国临床实践指南解读[J]. 中国康复理论与实践, 2010, 16(1): 83-86.
[6] 梁羽,李灿,罗兰,等. 脊髓损伤后早期小鼠肠道功能的变化[J]. 中国康复理论与实践, 2015, 21(2): 148-152.
[7] D'Agostino L, Ciacci C, Capuano G, et al. Metabolic fate of plasma diamine oxidase: evidence of isolated and perfused rat liver uptake [J]. Digestion, 1986, 34(4): 243-250.
[8] Cakmaz R, Büyükaşık O, Kahramansoy N, et al. A combination of plasma DAO and citrulline levels as a potential marker for acute mesenteric ischemia [J]. Libyan J Med, 2013, 8: 1-6.
[9] Luk GD, Bayless TM, Baylin SB. Diamine oxidase (histaminase). A circulating marker for rat mucosal maturation and integrity [J]. J Clin Invest, 1980, 66(1): 66-70.
[10] Kusche J, Lorenz W, Schmidt J. Oxidative deamination of biogenic amines by intestinal amine oxidases: histamine is specially inactivated by diamine oxidase [J]. Physiol Chem, 1975, 356(10): 1485-1486.
[11] Baylin SB, Stevens SA, Shakir KM. Association of diamine oxidase and ornithine with maturing cells in rapidly proliferating epithelium [J]. Biochim Biophys Acta, 1978, 541(3): 415-419.
[12] Fukudome I, Kobayashi M, Dabanaka K, et al. Diamine oxidase as a marker of intestinal mucosal injury and the effect of soluble dietary fiber on gastrointestinal tract toxicity after intravenous 5-fluorouracil treatment in rats [J]. Med Mol Morphol, 2014, 47(2): 100-107.
[13] Moriyama K, Kouchi Y, Morinaga H, et al. Diamine oxidase, a plasma biomarker in rats to GI tract toxicity of oral fluorouracil anti-cancer drugs [J]. Toxicology, 2006, 217(2-3): 233-239.
[14] Miyoshi J, Miyamoto H, Goji T, et al. Serum diamine oxidase activity as a predictor of gastrointestinal toxicity and malnutrition due to anticancer drugs [J]. J Gastroenterol Hepatol, 2015, 30(11): 1582-1590.
[15] 林丽芳. 不完全性脊髓损伤患者肠道功能评估与重建[J]. 当代护士, 2014(6): 50-51.
[16] 胡菊华. 电话追踪访视普外科病人存在的问题与对策[J]. 南方护理学报, 2005, 12(3): 90-91.
[17] 刘浩,白春宏. 脊髓损伤病人肠道管理[J]. 创伤外科杂志, 2014, 16(1): 79-81.
[18] 冯桂莲,李佩霞. 脊髓损伤患者肠道康复护理浅识[J]. 实用中医药杂志, 2010, 26(5): 342-343.
[19] 李静. 脊髓损伤患者神经源性肠道阶段化整体管理模式的建立及效果评价[J]. 中国当代医药, 2012, 19(4): 5-9.
[20] 叶水林,黄怀,沈彤,等. 早期康复介入对脊髓损伤患者膀胱肠道功能的影响[J]. 长春中医药大学学报, 2014, 30(4): 690-692.
[21] 李轲. 25例脊髓损伤患者的肠道功能训练分析[J]. 中国现代医生, 2015, 53(17): 85-91.
[22] Krassioukov A, Biering-Sorensen CF, Donovan W, et al. International Standards to Document Remaining Autonomic Function after Spinal Cord Injury (ISAFSCI), first edition 2012 [J]. Top Spinal Cord Inj Rehabil, 2012, 18(3): 282-296.
[23] Hou S, Rabchevsky AG. Autonomic consequences of spinal cord injury [J]. Compr Physiol, 2014, 4(4): 1419-1453.
[24] Luk GD, Bayless TM, Baylin SB. Plasma postheparin diamine oxidase. Sensitive provocative test for quantitating length of acute intestinal mucosal injury in the rat [J]. J Clin Invest, 1983, 71(5): 1308-1315.
[25] Honzawa Y, Nakase H, Matsuura M, et al. Clinical significance of serum diamine oxidase activity in inflammatory bowel disease: importance of evaluation of small intestinal permeability [J]. Inflamm Bowel Dis, 2011, 17(2): 23-25.
[26] Hilsden RJ, Meddings JB, Hardin J, et al. Intestinal permeability and postheparin plasma diamine oxidase activity in the prediction of Crohn's disease relapse [J]. Inflamm Bowel Dis, 1999, 5(2): 85-91.
[27] Cai C, Li W, Chen J, et al. Diamine oxidase as a marker for diagnosis of superior mesenteric arterial occlusion [J]. Hepatogastroenterology, 2012, 59(113): 155-158.
[28] Karabulut KU, Narci H, Gul M, et al. Diamine oxidase in diagnosis of acute mesenteric ischemia [J]. Am J Emerg Med, 2013, 31(2): 309-312.
[29] Seidl H, Gundling F, Pehl C, et al. Multiple intraluminal impedance measurement and manometry in the jejunum of healthy subjects: a new tool for diagnosis of small bowel motility [J]. Digestion, 2010, 81(3): 172-180.
[30] Chang IY, Glasgow NJ, Takayama I, et al. Loss of interstitial cells of Cajal and development of electrical dysfunction in murine small bowelobstruction [J]. J Physiol, 2001, 536(2): 555-568.
[31] Lee JC, Thuneberg L, Berezin I, et al. Generation of slow waves in membrane potential is an intrinsic property of interstitial cell of Cajal [J]. Am J Physiol, 1999, 40(2): 409-423.
[32] Ordög T, Ward SM, Sanders KM. Interstitial cells of cajal generate electrical slow waves in the murine stomach [J]. J Physiol, 1999, 518(Pt 1): 257-269.
