Jiang Guo-Tao, Chen Xiao, Li Dong, An Hui-Xia, Jiao Jun-Dong
Department of Nephrology, Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China.
Mol Med Rep. 2014 Sep;10(3):1501-8. doi: 10.3892/mmr.2014.2323. Epub 2014 Jun 13.
The aim of the present study was to examine the protective effects of the urinary trypsin inhibitor ulinastatin (UTI) on renal interstitial inflammation and fibrosis in rats subjected to unilateral ureteral obstruction (UUO). A total of 24 male Wistar rats were randomly divided into the three groups; the sham operation (SOR) group (n=8), the UUO group (n=8) and the UUO+UTI group (post‑UUO UTI treatment, n=8). UUO was performed with complete ligation of the left ureter. As a medical intervention, saline (4 ml kg‑1 d‑1) and UTI (40000 units kg‑1 d‑1) were injected, respectively, into the animals of the corresponding groups on day one following surgery. The rats in all three groups were euthanized on day seven post surgery. Blood samples were harvested for blood urea nitrogen (BUN) and serum creatinine (Scr) content measurements. The degree of interstitial pathological changes in the tissues from the obstructed kidneys were observed through hematoxylin and eosin (H&E) and Masson staining. The CD68+ macrophage amount, tumor necrosis factor‑α (TNF‑α), interleukin 1β (IL‑1β), nuclear factor‑κB (NF‑κB), transforming growth factor‑β1 (TGF‑β1) and type I collagen (Col‑I) levels were examined immunohistochemically. The protein expression levels of NF‑κB were examined using western blot analysis. Total superoxide dismutase (SOD) activity and malondialdehyde (MDA) content of homogenates were measured spectrophotometrically. The results revealed that ulinastatin had no statistically significant effect on the BUN and Scr levels (P>0.05). However, in comparison with the SOR group, the UUO group exhibited significantly more severe renal interstitial pathological injury in terms of tubular dilation, epithelial atrophy, renal interstitial inflammatory cell infiltration and proliferation of fibrous tissues, as well as significantly elevated levels of interstitial CD68+ macrophages, IL‑1β, TNF‑α, NF‑κB, TGF‑β1 and Col‑I (P<0.01). UTI treatment significantly reduced UUO‑induced renal interstitial damage with reduced levels of interstitial CD68+ macrophages, IL‑1β, TNF‑α, NF‑κB, TGF‑β1 and Col‑I and MDA (P<0.05), and increased SOD levels (P<0.05). In conclusion, the present study indicated that UTI is able to effectively inhibit UUO‑side renal interstitial inflammatory reaction and fibrosis in UUO‑inflicted rats.
本研究旨在探讨尿胰蛋白酶抑制剂乌司他丁（UTI）对单侧输尿管梗阻（UUO）大鼠肾间质炎症和纤维化的保护作用。将24只雄性Wistar大鼠随机分为三组：假手术（SOR）组（n = 8）、UUO组（n = 8）和UUO + UTI组（UUO术后UTI治疗，n = 8）。通过完全结扎左输尿管进行UUO手术。作为医学干预措施，术后第1天分别向相应组的动物注射生理盐水（4 ml·kg⁻¹·d⁻¹）和UTI（40000单位·kg⁻¹·d⁻¹）。术后第7天对三组大鼠实施安乐死。采集血样用于测量血尿素氮（BUN）和血清肌酐（Scr）含量。通过苏木精-伊红（H&E）染色和Masson染色观察梗阻肾脏组织的间质病理变化程度。采用免疫组织化学方法检测CD68⁺巨噬细胞数量、肿瘤坏死因子-α（TNF-α）、白细胞介素1β（IL-1β）、核因子-κB（NF-κB）、转化生长因子-β1（TGF-β1）和I型胶原蛋白（Col-I）水平。使用蛋白质印迹分析检测NF-κB的蛋白表达水平。通过分光光度法测量匀浆中总超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量。结果显示，乌司他丁对BUN和Scr水平无统计学显著影响（P > 0.05）。然而，与SOR组相比，UUO组在肾小管扩张、上皮萎缩、肾间质炎性细胞浸润和纤维组织增生方面表现出明显更严重的肾间质病理损伤，并且间质CD68⁺巨噬细胞、IL-1β、TNF-α、NF-κB、TGF-β1和Col-I水平显著升高（P < 0.01）。UTI治疗显著减轻了UUO诱导的肾间质损伤，使间质CD68⁺巨噬细胞、IL-1β、TNF-α、NF-κB、TGF-β1和Col-I以及MDA水平降低（P < 0.05），并使SOD水平升高（P < 0.05）。总之，本研究表明UTI能够有效抑制UUO大鼠患侧肾间质炎症反应和纤维化。
