骨髓间充质干细胞通过 TNF-α 调节机制减少大鼠肠缺血再灌注损伤、ZO-1 下调和紧密连接破坏。
Bone-marrow mesenchymal stem cells reduce rat intestinal ischemia-reperfusion injury, ZO-1 downregulation and tight junction disruption via a TNF-α-regulated mechanism.
机构信息
Department of Organ Transplantation, Tianjin First Central Hospital, Tianjin 300192, China.
出版信息
World J Gastroenterol. 2013 Jun 21;19(23):3583-95. doi: 10.3748/wjg.v19.i23.3583.
AIM
To investigate the effect of bone-marrow mesenchymal stem cells (BM MSCs) on the intestinal mucosa barrier in ischemia/reperfusion (I/R) injury.
METHODS
BM MSCs were isolated from male Sprague-Dawley rats by density gradient centrifugation, cultured, and analyzed by flow cytometry. I/R injury was induced by occlusion of the superior mesenteric artery for 30 min. Rats were treated with saline, BM MSCs (via intramucosal injection) or tumor necrosis factor (TNF)-α blocking antibodies (via the tail vein). I/R injury was assessed using transmission electron microscopy, hematoxylin and eosin (HE) staining, immunohistochemistry, western blotting and enzyme linked immunosorbent assay.
RESULTS
Intestinal permeability increased, tight junctions (TJs) were disrupted, and zona occludens 1 (ZO-1) was downregulated after I/R injury. BM MSCs reduced intestinal mucosal barrier destruction, ZO-1 downregulation, and TJ disruption. The morphological abnormalities after intestinal I/R injury positively correlated with serum TNF-α levels. Administration of anti-TNF-α IgG or anti-TNF-α receptor 1 antibodies attenuated the intestinal ultrastructural changes, ZO-1 downregulation, and TJ disruption.
CONCLUSION
Altered serum TNF-α levels play an important role in the ability of BM MSCs to protect against intestinal I/R injury.
目的
研究骨髓间充质干细胞（BM MSCs）对缺血再灌注（I/R）损伤肠黏膜屏障的影响。
方法
通过密度梯度离心法从雄性 Sprague-Dawley 大鼠中分离 BM MSCs，培养后通过流式细胞术进行分析。通过阻断肠系膜上动脉 30 分钟来诱导 I/R 损伤。用生理盐水、BM MSCs（黏膜内注射）或肿瘤坏死因子（TNF）-α 阻断抗体（尾静脉注射）处理大鼠。使用透射电子显微镜、苏木精和伊红（HE）染色、免疫组织化学、蛋白质印迹和酶联免疫吸附试验评估 I/R 损伤。
结果
I/R 损伤后肠通透性增加，紧密连接（TJs）破坏，闭合蛋白 1（ZO-1）下调。BM MSCs 减少了肠黏膜屏障的破坏、ZO-1 下调和 TJ 破坏。肠 I/R 损伤后的形态学异常与血清 TNF-α 水平呈正相关。给予抗 TNF-α IgG 或抗 TNF-α 受体 1 抗体可减轻肠超微结构变化、ZO-1 下调和 TJ 破坏。
结论
血清 TNF-α 水平的改变在 BM MSCs 保护肠 I/R 损伤的能力中发挥重要作用。
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