解剖学报

解剖学报 ›› 2014, Vol. 45 ›› Issue (3): 405-410.doi: 10.3969/j.issn.0529-1356.2014.03.020

• 组织学胚胎学发育生物学 • 上一篇    下一篇

输尿管梗阻及再通大鼠肾组织中单核-巨噬细胞相关因子动态表达

陆红1 杨嫆嫆1 梁勇2 林成成2 洪炜龙2 陈必成2 白永恒2*   

  1. 1. 温州医科大学附属第一医院医学检验中心，浙江 温州 325000; 2. 温州医科大学附属第一医院外科实验室， 浙江 温州 325000
  • 收稿日期:2013-10-08 修回日期:2013-11-14 出版日期:2014-06-06 发布日期:2014-06-06
  • 通讯作者: 白永恒 E-mail:greatsailor@163.com
  • 基金资助:

    浙江省自然科学基金;温州市科技计划项目

Expression of monocyte-macrophage-related factors in kidney tissues of rats with ureter obstruction and recanalization

LU Hong1 YANG Rong-rong1 LIANG Yong2 LIN Cheng-cheng2 HONG Wei-long2 CHEN Bi-cheng2 BAI Yong-heng 2*   

  1. 1. Department of Laboratory Medicine, the First Affiliated Hospital of Wenzhou Medical University, Zhejiang Wenzhou 325000, China;2.Wenzhou Key Laboratory of Surgery, the First Affiliated Hospital of Wenzhou Medical University, Zhejiang Wenzhou 325000, China
  • Received:2013-10-08 Revised:2013-11-14 Online:2014-06-06 Published:2014-06-06
  • Contact: BAI Yong-heng E-mail:greatsailor@163.com

摘要：

目的 探讨输尿管梗阻及再通大鼠肾间质纤维化发生和恢复过程中单核-巨噬细胞相关因子的动态表达。方法 将48只SD大鼠随机分成梗阻和再通两个实验部分，梗阻部分分为假手术组（sham，n=6）、单侧输尿管梗阻（UUO）3d（n=6）、UUO 7d（n=6）和UUO 14d（n=6）；再通部分分为双侧输尿管梗阻（RBUO）0d（n=6）、RBUO后再通3d（n=6）、RBUO后再通 7d（n=6）和RBUO 后再通14d（n=6）。术后3、7和14 d后处死取其肾脏组织。采用HE和Masson染色观察肾组织病理改变和间质纤维化程度；免疫组织化学染色检测肾组织内单核细胞趋化蛋白-1（MCP-1）、巨噬细胞克隆刺激因子（M-CSF）和活化巨噬细胞标志物CD68的表达水平；Real-time PCR检测MCP-1和M-CSF mRNA表达水平；ELISA测定TGF-β1含量。 结果 与Sham大鼠相比，UUO大鼠随着梗阻时间延长，纤维化程度加剧。同时TGF-β1水平明显升高。输尿管再通后，纤维化程度随时间延长明显减轻，且TGF-β1水平明显下调。UUO大鼠肾组织中，MCP-1和M-CSF mRNA和表达蛋白随梗阻时间延长而增加；梗阻再通后，MCP-1和M-CSF mRNA和蛋白表达随再通时间延长持续下降。MCP-1和M-CSF在UUO或再通大鼠肾组织中的表达与CD68呈现一致性变化趋势。 结论 输尿管梗阻后，M-CSF和MCP-1的动态表达反映单核-巨噬细胞的活化和聚集状态，这可能是间质纤维化发生十分重要的炎症基础。而输尿管再通可抑制活化和趋化的单核-巨噬细胞，控制炎症反应，缓解肾间质纤维化。

关键词: 单核-巨噬细胞, 趋化因子, 输尿管梗阻, 再通, 纤维化, 免疫组织化学, 实时定量聚合酶链反应, 大鼠

Abstract:

Objective To investigate the expression of monocyte-macrophage-related factors and interstitial fibrosis in kidney tissues of rats with ureter obstruction and recanalization. Methods Forty-eight male Spragur-Dawley rats were divided randomly into the obstructive group: sham (n=6), unilateral ureteral obstruction（UUO）3 days (n=6), UUO 7 days (n=6), and UUO 14 days (n=6) and recanalization group: bilateral ureteral obstruction（RBUO）0 day (n=6), 3 days after RBUO (n=6), 7 days after RBUO (n=6), and 14 days after RBUO (n=6). The kidneys were excised on day 3, 7, and 14, and the deposition of collagen fibers in kidney was detected with HE and Masson staining. Immunohistochemical analysis was performed to evaluate the protein expressions of monocyte chemoattractant protein-1 (MCP-1), macrophage colony-stimulating factor (M-CSF) and activated-macrophage marker CD68. Real-time PCR was used to detect the mRNA expressions of MCP-1 and M-CSF. TGF-β1 levels were determined by ELISA. Results Fibrosis observed with HE and Masson staining was obviously increased in kidney tissue of UUO rats, and aggravated as time prolonged, but alleviated in rats with recanalization. TGF-β1 levels were increased obviously in the UUO group, but decreased in rats with recanalization compared with those in BUO rats. In UUO rats, mRNA and protein expression levels of MCP-1 and M-CSF were increased. MCP-1 and M-CSF expression was gradually decreased in rats with recanalization compared with those in BUO rats. The dynamic change in expression of MCP-1 and M-CSF in both UUO rats and recanalization rats was consistent with the change in expression of CD68. Conclusion Dynamic change in expression of MCP-1 and M-CSF in kidney tissues reflects change of activated and accumulated monocyte-macrophages, which may be one of the major mechanisms contributing to fibrosis induced by ureter obstruction. Renal fibrosis is alleviated by down-regulated expression of monocyte-macrophages factors with recanalization operation.

Key words: Monocyte-macrophage, Chemoattractant factor, Ureter obstruction, Recanalization, Fibrosis, Immunohistochemistry, Real-time PCR, Rat