他克莫司对小鼠肠黏膜屏障功能的损伤作用及分子机制

发布时间：2018-11-13 07:47

【摘要】：目的观察他克莫司(FK506)对小鼠肠黏膜屏障功能的损伤作用,并探讨其分子机制。方法将24只C57BL/6小鼠随机分为高剂量组、低剂量组和对照组各8只,分别予FK506 5 mg/kg、FK506 1 mg/kg、生理盐水灌胃。用药30 d处死小鼠,收集肠黏膜组织标本,采用HE染色法观察肠黏膜组织结构变化,透射电镜下观察肠黏膜上皮细胞紧密连接的超微结构改变,采用分光光度法测定血浆D-乳酸,氧电极法测定肠组织线粒体呼吸功能,高压液相色谱法检测肠黏膜上皮细胞线粒体三磷酸腺苷(ATP)酶活性。结果低、高剂量组小鼠体质量较对照组明显减轻,高剂量组减轻更明显(P均0.05);低、高剂量组小鼠肠上皮细胞间紧密连接超微结构遭到破坏,其损伤程度与FK506剂量呈正相关;与对照组比较,低、高剂量组小鼠血浆D-乳酸水平升高,高剂量组升高更明显(P均0.05);与对照组比较,低、高剂量组小鼠小肠3态呼吸耗氧量、呼吸控制率、膜电位、ATP酶活性下降,4态呼吸耗氧量升高,高剂量组变化更明显(P均0.05)。结论 FK506能损伤小鼠肠黏膜屏障功能,且损伤程度呈剂量依赖性;其分子机制可能是通过损伤小鼠肠黏膜细胞线粒体功能而损伤肠黏膜屏障功能。
[Abstract]:Objective to observe the effects of tacrolimus (FK506) on intestinal mucosal barrier function in mice and explore its molecular mechanism. Methods Twenty-four C57BL/6 mice were randomly divided into three groups: high dose group, low dose group and control group. They were given FK506 5 mg/kg,FK506 1 mg/kg, saline respectively. 30 days after administration, the mice were killed and the intestinal mucosal tissue samples were collected. The changes of intestinal mucosal structure were observed by HE staining, and the ultrastructural changes of the tight junction of intestinal mucosal epithelial cells were observed under transmission electron microscope (TEM). Plasma D-lactic acid was measured by spectrophotometry, mitochondrial respiratory function of intestinal tissue was measured by oxygen electrode method, and mitochondrial adenosine triphosphate (ATP) activity of intestinal mucosal epithelial cells was measured by high pressure liquid chromatography (HPLC). The results showed that the body weight of the mice in the high dose group was significantly lower than that in the control group, and that in the high dose group was significantly lower than that in the control group (P 0.05). The ultrastructure of tight junction between intestinal epithelial cells was destroyed in the low and high dose groups, and the degree of injury was positively correlated with the dose of FK506. Compared with the control group, the plasma D-lactic acid level in the low and high dose groups was higher than that in the high dose group (P 0.05). Compared with the control group, the oxygen consumption, respiratory control rate, membrane potential and ATP enzyme activity of the small intestine in the low and high dose groups were decreased, and the oxygen consumption in the four states were increased, especially in the high dose group (all P 0. 05). Conclusion FK506 can damage the intestinal mucosal barrier function in a dose-dependent manner, and its molecular mechanism may be to damage the intestinal mucosal barrier function by damaging the mitochondrial function of the mouse intestinal mucosal cells.
【作者单位】：青岛大学附属医院;南京军区南京总医院;
【基金】：山东省优秀中青年科学家科研奖励基金项目(BS2011YY004) 黎介寿院士肠屏障研究专项基金项目(LJS201105)
【分类号】：R965

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