天津医药 ›› 2015, Vol. 43 ›› Issue (3): 252-255.doi: 10.11958/j.issn.0253-9896.2015.03.008

• 实验研究 • 上一篇    下一篇

谷氨酰胺预处理对大鼠肠缺血再灌注损伤的保护作用及其对 eNOS-NO 通路的影响#br#

牛琼, 王爱丽, 王伟, 胡营滨, 刘成霞   

  1. 山东省滨州市滨州医学院附属医院消化内科 (邮编 256603
  • 收稿日期:2014-09-30 修回日期:2014-12-31 出版日期:2015-03-15 发布日期:2015-03-15
  • 通讯作者: 刘成霞 E-mail:phdlcx@163.com
  • 作者简介:牛琼 (1973), 女, 硕士, 副教授, 副主任医师, 主要从事肠黏膜屏障的临床与基础研究
  • 基金资助:
    山东省自然科学基金 (ZR2010HL051

The protective effect of glutamine pretreatment on intestinal ischemia-reperfusion injury and eNOS/NO levels in rats#br# #br#

NIU QiongWANG AiliWANG WeiHU YingbinLIU Chengxia   

  1. Department of Gastroenterology, the Affiliated Hospital of Binzhou Meidcal University, Binzhou 256603, China
  • Received:2014-09-30 Revised:2014-12-31 Published:2015-03-15 Online:2015-03-15
  • Contact: LIU Chengxia E-mail:phdlcx@163.com

摘要: 目的 探讨谷氨酰胺 (Gln) 预处理对大鼠肠缺血再灌注损伤 (IR) 后的保护作用及其对内皮型一氧化氮合酶eNOS/一氧化氮 (NO) 信号通路的影响。方法 30 只健康雄性 Wistar 大鼠随机分为假手术 (Sham) 组、 IR 组、Gln 组, 每组 10 只, Gln 组给予谷氨酰胺 1 g/ kg·d), 连续灌胃 7 dSham 组和 IR 组以同等剂量生理盐水灌胃 7 dSham 组仅分离肠系膜上动脉(SMA)而根部不夹闭。IR 组和 Gln 组均用无损伤血管夹夹闭 SMA 根部, 30 min 后放松血管夹形成再灌注损伤模型。各组大鼠均于制模后 24 h 采集腹主动脉血和回肠标本。应用 HE 染色法观察肠黏膜组织形态学改变, ELISA 试剂盒双抗体夹心法测定 D-乳酸、 内毒素含量, 硝酸还原酶法检测血清 NO 的含量, 比色法检测血清 eNOS、 诱导型一氧化氮合酶 (iNOS) 的含量, 荧光定量 PCR RT-PCR) 检测大鼠肠组织 eNOSiNOS mRNA表达水平。结果 再灌注后, 与 Sham 组相比, IR 组肠黏膜绒毛上皮脱落, 固有层崩解, 部分绒毛顶端出血, 腺体明显受损; Gln 组表现为肠黏膜绒毛上皮下间隙扩大, 但不明显, 绒毛轻度水肿, 腺体大致正常, 固有层轻度水肿。IR 组血D-乳酸、 内毒素、 iNOS 水平及肠组织 iNOS mRNA 表达水平均高于 Sham 组和 Gln 组(P < 0.05)。IR 组血清 NOeNOS 含量及组织中 eNOS mRNA 表达量均低于 Sham 组和 Gln 组(P < 0.05)。结论 谷氨酰胺预处理可以减轻肠缺血再灌注后的组织形态学改变及损伤, 其机制可能与抑制 iNOS 表达, 增加 eNOS 表达, 从而增加 NO 活性有关。

关键词: 谷氨酰胺, 肠, 缺血, 再灌注损伤, 一氧化氮合酶, 肠缺血再灌注损伤, 内皮型一氧化氮合酶, 诱导型一氧化氮合酶

Abstract: Objective To investigate the protective effect of glutamine(Gln) pretreatment on intestinal ischemia-reperfusion (I/R) injury and endothelial nitric oxide synthase (eNOS) / nitric oxide (NO) signaling pathway in rat model. Methods Thirty male Wistar rats were randomly divided into three groups(n=10 for each group): sham group, I/R group and Gln group. Animals were pretreated with 1 g/ kg·dGln by orogastric route for 7 days in Gln group, and normal saline was given to the other two groups in the same dose. Intestinal I/R was induced by 30 min occlusion of the superior mesenteric artery followed by 24 h of reperfusion. After the operation, the intestinal histopathological changes, the plasma endotoxin level, serum D-lactic acid, eNOS, inducible NOS(iNOS)activity and NO levels were detected by ultraviolet spectrophotometer. The mRNA expressions of myocardial eNOS and iNOS were detected by real-time fluorescence quantitative PCR (RT-PCR). Results After reperfusion, in IR group, extensive epithelial sloughing and mucosal ulceration of villous tips were observed, whereas these findings did not occur in Gln group and sham group. Compared with IR group, the serum NO, eNOS levels and eNOS mRNA expression of intestinal tissue were elevated in Gln group (P < 0.01), but the plasma endotoxin level, serum D-lactic acid, serum iNOS and intestinal iNOS mRNA expression decreased in IR group(P < 0.05). Conclusion Glutamine pretreatment has protective effects on intestinal ischemia-reperfusion injury in vivo. The mechanism may be related to the inhibition of iNOS expression and the increased expression of eNOS, thereby increasing NO activity.

Key words: glutamine, intestines, ischemia, reperfusion injury, nitric oxide synthase, intestinal ischemia-reperfusion injury, eNOS, iNOS