Department of Nephropathy, Henan Provincial Hospital of Traditional Chinese Medicine, Zhengzhou, Henan, China (mainland).
Henan University of Chinese Medicine, Second Clinical Medical College, Zhengzhou, Henan, China (mainland).
Med Sci Monit. 2020 May 30;26:e921894. doi: 10.12659/MSM.921894.
BACKGROUND This study aimed to investigate the effects of hirudin on the production of extracellular matrix (ECM) factors by renal tubular epithelial cells in a rat model of diabetic kidney disease (DKD) and HK-2 human renal tubule epithelial cells. MATERIAL AND METHODS Sprague-Dawley rats were divided into the normal control group (n=10), the normal control+hirudin group (n=10), the DKD model group (n=12) and the DKD+hirudin group (n=12). At the end of the study, renal histopathology was undertaken, and the expression of type IV collagen, fibronectin, hypoxia-inducible factor-1alpha (HIF-1alpha), and vascular endothelial growth factor (VEGF) were evaluated using immunohistochemistry, Western blot, and quantitative real-time polymerase chain reaction (qRT-PCR). HK-2 cells were cultured in glucose and treated with hirudin. Protein and mRNA expression of fibronectin, type IV collagen, HIF-1alpha, and VEGF were evaluated following knockdown or overexpression of HIF-1alpha. RESULTS Hirudin significantly improved renal function in the rat model of DKD (P<0.01), and significantly down-regulated the expression of fibronectin, type IV collagen, HIF-1alpha, and VEGF proteins (P<0.05). The expression of ECM associated proteins was increased in HK-2 cells treated with high glucose and reduced in the high glucose+shRNA HIF-1alpha group (P<0.05). Compared with the control group, the expression of ECM associated proteins was increased in the HIF-1alpha over-expressed group, and decreased following treatment with hirudin (P<0.05). CONCLUSIONS Hirudin reduced the expression of markers of ECM by inhibiting the HIF-1alpha/VEGF signaling pathway in DKD renal tubular epithelial cells.
本研究旨在探讨水蛭素对糖尿病肾病（DKD）大鼠模型及 HK-2 人肾小管上皮细胞中细胞外基质（ECM）因子产生的影响。
将 Sprague-Dawley 大鼠分为正常对照组（n=10）、正常对照组+水蛭素组（n=10）、DKD 模型组（n=12）和 DKD+水蛭素组（n=12）。研究结束时，进行肾脏组织病理学检查，并采用免疫组化、Western blot 和实时定量聚合酶链反应（qRT-PCR）评估 IV 型胶原、纤维连接蛋白、缺氧诱导因子-1α（HIF-1α）和血管内皮生长因子（VEGF）的表达。将 HK-2 细胞在葡萄糖中培养，并给予水蛭素处理。敲低或过表达 HIF-1α后，评估纤维连接蛋白、IV 型胶原、HIF-1α 和 VEGF 的蛋白和 mRNA 表达。
水蛭素显著改善 DKD 大鼠的肾功能（P<0.01），并显著下调纤维连接蛋白、IV 型胶原、HIF-1α和 VEGF 蛋白的表达（P<0.05）。高糖处理的 HK-2 细胞中 ECM 相关蛋白的表达增加，而高糖+shRNA HIF-1α组中则减少（P<0.05）。与对照组相比，HIF-1α过表达组 ECM 相关蛋白的表达增加，而给予水蛭素治疗后则减少（P<0.05）。
水蛭素通过抑制 DKD 肾小管上皮细胞中的 HIF-1α/VEGF 信号通路，减少 ECM 标志物的表达。
