上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

翠云草对单侧输尿管梗阻大鼠肾间质纤维化抑制作用研究

徐剑1，袁红伶1，倪凯2，周晓萍1，李嵘1，周艳1，高敏2   

  1. 1云南省第一人民医院，昆明理工大学附属昆华医院肾内科，昆明 650032；2云南省中医中药研究院，昆明 650032
  • 出版日期:2016-12-28 发布日期:2016-12-29
  • 通讯作者: 袁红伶，电子信箱: kmyuanhl@yeah.net。
  • 作者简介:徐剑（1978—），男，主治医师，硕士；电子信箱：xujian3979@sina.com。
  • 基金资助:

    国家自然科学基金（81460649）；云南省科技厅应用基础研究项目（2013FZ）

Study on the inhibitory effect of Herba Selaginellae Uncinatae on renal interstitial fibrosis in rats with unilateral ureteral obstruction

XU Jian1, YUAN Hong-ling1, NI Kai2, ZHOU Xiao-ping1, LI Rong1, ZHOU Yan1, GAO Min2   

  1. 1 Renal Department of Internal Medicine, First People's Hospital of Yunnan Province, Kunhua Hospital Affiliated to Kunming University of Science and Technology, Kunming 650032,China;2 Yunnan Institute of Traditional Chinese Medicine, Kunming 650032, China

  • Online:2016-12-28 Published:2016-12-29
  • Supported by:

    National Natural Science Foundation of China, 81460649; Applied Basic Research Project of Science and Technology Department of Yunnan Province, 2013FZ

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摘要：

目的 ·研究翠云草对单侧输尿管梗阻（UUO）大鼠肾间质纤维化的抑制作用，以及可能的机制。方法 ·108只SD大鼠，随机均分为假手术组、模型组、低浓度药物组和高浓度药物组。模型组、低浓度药物组和高浓度药物组先建立左侧输尿管梗阻模型，低浓度药物组（2 g/kg）和高浓度药物组（4 g/kg）给予翠云草提取液持续灌胃14 d。分别于建模后3、7、14 d，每组取9只，制备肾组织切片，通过苏木精-伊红染色（H-E染色）、Masson染色，观察UUO大鼠模型及治疗组肾脏病理组织学改变，以及肾脏纤维化情况。采用免疫组织化学法检测各组大鼠α-平滑肌肌动蛋白（α-SMA）、转化生长因子β1（TGF-β1）、单核细胞趋化蛋白1（MCP-1）在肾组织的表达情况；通过RT-PCR法定量检测α-SMA、TGF-β1和MCP-1 mRNA水平。结果 ·模型组大鼠术后3 d时，肾脏病理改变主要以肾间质小管炎症细胞浸润为主，可见肾小管扩张，肾间质纤维化不明显，肾小球正常；术后14 d时，部分肾小管萎缩、排列紊乱，肾间质可见弥漫性炎症细胞浸润，纤维化程度严重。2个药物治疗组同模型组比较，炎症细胞浸润程度、肾小管的萎缩程度及纤维化程度均较轻。免疫组织化学检测结果显示：α-SMA、TGF-β1和MCP-1均沿肾小管间质、小血管壁分布，术后14 d时，模型组染色最深，2个药物治疗组的平均光密度均显著低于模型组（均P<0.05）。RT-PCR结果显示，2个药物治疗组肾组织的α-SMA、TGF-β1和 MCP-1 mRNA水平均较模型组显著下调（均P<0.05）。结论 ·翠云草可下调UUO大鼠肾组织α-SMA、TGF-β1和MCP-1的水平，减少炎症细胞浸润，抑制肾纤维化进展。

关键词: 翠云草, 单侧输尿管梗阻大鼠, 肾间质纤维化, 抑制作用

Abstract:

Objective · To investigate the inhibitory effect of Herba Selaginellae Uncinatae (HSU) on renal interstitial fibrosis in rats with unilateral ureteral obstruction (UUO) and possible mechanisms. Methods · One hundred and eight Sprague-Dawley rats were randomly assigned to the sham-operation group, the model group, the low-dose HSU group, and the high-dose HSU group. The model of left ureteral obstruction was built in the latter three groups. The low-dose HSU group and the high-dose HSU group received lavage with 2 and 4 g/kg of HSU extract for 14 days, respectively. Renal tissue sections from nine rats in each group were prepared 3, 7, and 14 days,respectively, after the model was built. H-E staining and Masson staining were used to observe renal histopathological changes and renal fibrosis. Immunohistochemistry was used to measure the expressions of MCP-1, TGF-β1, and α-smooth muscle actin (α-SMA) in renal tissues. RT-PCR was used to quantitatively measure mRNA levels of TGF-β1, α-SMA, and MCP-1. Results · Renal pathological changes in the model group 3 days after operation were mainly inflammatory cell infiltration with tubular dilation, insignificant renal interstitial fibrosis, and normal glomeruli. Some renal tubules were atrophic with disorganized arrangement 14 days after operation. Diffuse inflammatory cell infiltration was observed in renal interstitium with severe fibrosis. Compared with the control group, two HSU groups had less inflammatory cell infiltration, renal tubular atrophy, and fibrosis. The immunohistochemical results showed that α-SMA, MCP-1, and TGF-β1 distributed in renal tubular interstitium and on small vessel walls. The normal saline group had the darkest staining 14 days after operation. The optical density was significantly lower in two HSU groups than in the control group (P<0.05). RT-PCR results showed that mRNA levels of TGF-β1, α-SMA, and MCP-1 in renal tissues were significantly lower in two HSU groups than in the control group (P<0.05). Conclusion · HSU can down-regulate α-SMA, TGF-β1, and MCP-1 levels in renal tissues of UUO rats, reduce inflammatory cell infiltration, and inhibit the progression of renal fibrosis.

Key words: Herba Selaginellae Uncinatae, UUO rat, renal interstitial fibrosis, inhibition