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肾脏病与透析肾移植杂志

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尿毒症大鼠肠道屏障功能紊乱与微炎症的关系

  

  • 出版日期:2019-10-28 发布日期:2020-01-07

Intestinal barrier dysfunction and relationship with microinflammation in  uremic rats#br#

  • Online:2019-10-28 Published:2020-01-07

摘要: 目的:评估尿毒症肠道屏障功能状态,探讨肠道屏障功能紊乱与微炎症状态的关系。
方法:SD大鼠随机分成尿毒症组和假手术组,5/6肾切除法建立尿毒症模型,假手术组仅打开肾包膜。构建绿色荧光蛋白标记的示踪菌并饲喂两组大鼠,应用PCR法从肝、脾、肠系膜淋巴结、外周血等肠外组织中扩增绿色荧光蛋白特异性基因片段,以检测是否存在肠道细菌移位至该肠外组织位点;HE染色观察肠道组织病理变化;透射电镜观察超微结构变化;免疫组化法观察肠组织JAM1、claudin1和Occludin表达;免疫比浊法检测血高敏C反应蛋白(hsCRP)水平、酶联免疫吸附试验检测血中炎症因子白细胞介素6(IL6)、肿瘤坏死因子α(TNFα)水平。
结果:尿毒症组肠道细菌移位率(70%)显著高于假手术组(10%)(P<005)。超微结构显示尿毒症组肠上皮细胞间紧密连接结构不完整,微绒毛走行紊乱,部分断裂、消失。与假手术组相比,尿毒症组血hsCRP(603±170 mg/L vs 345±126  mg/L)、IL6(3762±321 pg/ml vs 1981±655  pg/ml)、TNFα(2848±1085  pg/ml vs 1733±572  pg/ml)均显著升高(P<005)。
结论:尿毒症大鼠存在肠道屏障功能障碍,肠道细菌可移位进入远隔脏器及血液循环并激活炎症反应,可能是尿毒症发生微炎症状态的机制之一。

关键词: 肠道屏障, 尿毒症, 微炎症, 细菌移位

Abstract: Objective:To evaluate the status of intestinal barrier function and to explore the relationship between intestinal barrier dysfunction and microinflammation in  uremic rats.
Methodology:SD rats were randomly divided into uremic group and sham operation group.The uremic model was established by 5/6 nephrectomy.Only renal capsule were opened in sham operation group.The green fluorescent proteinlabeled tracer was constructed and fed to two groups of rats.The green fluorescent proteinspecific gene fragment was amplified from the extraintestinal tissues such as liver,spleen,mesenteric lymph node and peripheral blood by PCR to detect the presence of intestinal bacteria were transferred to the extraintestinal tissue site; HE staining was used to observe pathological changes of intestinal tissue,transmission electron microscopy was used to observe ultrastructural changes.Expressions of JAM1,claudin1 and Occludin in intestinal tissues were observed by immunohistochemistry.Serum hsCRP,IL6 and TNFα levels were detected.
Results:The intestinal bacterial translocation rate  was significantly higher in uremic group (70% vs 10%,P<005).The ultrastructure showed that the tight junction structure between intestinal epithelial cells in uremic group was incomplete and electron density decreased.The microvilli were disorganized,partially broken and disappeared.Serum hsCRP (603±170 vs 345±126),IL6 (3762±321 vs 1981±655),and TNFα (2848±1085 vs 1733±572) were significantly higher in uremic group than those in shamoperated group. (P<005).
Conclusion:
Intestinal mucosal barrier dysfunction exists in experimental uremia.Intestinal bacteria can translocate to distant organs and blood circulation,then activate inflammatory response,which may be one of the mechanisms of microinflammation.

Key words: intestinal barrier, uremia, microinflammation, bacterial translocation