Aims: To investigate the role of Vasohibin-1 (VASH-1), silence information adjustment factor 2-related enzyme 1 (SIRT1)/hypoxic-inducible factor 1α (HIF1α) and transforming growth factor-β1 (TGFβ1) /Smad3 signaling pathways in oxidative stress and fibrosis of diabetic kidney disease (DKD).

Materials and Methods: A diabetic rat model was established in vivo and rat mesangial cells (RMCs) were cultured in vitro with high glucose via transfection with Vash1 small interfering RNA (siRNA), Hif1a siRNA, Sirt1 siRNA and TGFβ1/Smad3 pathway inhibitor (SB431542). Renal histology was used to detect renal changes. Real-time PCR and western blot were used to analyze the expression of VASH-1, SIRT1, HIF1α, TGFβ1, Smad3, vascular endothelial growth factor (VEGF), connective tissue growth factor (CTGF) and fibronectin (FN). Expression levels of tumor necrosis factor-α (TNFα), TGFβ1, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX), and malondialdehyde (MDA) in rat tissues and cell culture supernatant were detected by ELISA and chemiluminescence assay, while cell proliferation was detected by CCK-8.

Results: The level of VASH-1 in renal tissues of diabetic rats was decreased, while both high glucose and Vash1 siRNA inhibited the expression of VASH-1 and SIRT1, increased the levels of HIF1α, TGFβ1, and Smad3 in RMCs, thus up-regulating oxidative stress and fibrosis factors, and abnormally increasing cell proliferation activity (P < 0.05). However, inhibition of SIRT1/HIF1α signaling pathway only reduced TGFβ1 and Smad3 (P < 0.05), while VASH-1 remained unchanged (P > 0.05).

Conclusion: VASH-1 was under-expressed in renal tissues of diabetic rats and regulated the pathological process of oxidative stress and fibrosis in DKD via downstream SIRT1/HIF1α and TGFβ1/Smad3 signaling pathways.

目的： 研究Vasohibin-1（VASH-1），沉默信息调节因子2相关酶1（SIRT1）/低氧诱导因子1α（HIF1α）和转化生长因子β1（TGFβ1）/ Smad3信号通路在氧化中的作用糖尿病肾病（DKD）的应激和纤维化。

材料和方法： 建立了糖尿病大鼠模型 体内 和大鼠肾小球系膜细胞（RMCs）培养 体外 通过转染高糖 Vash1 小干扰RNA（siRNA）， Hif1a siRNA， Sirt1siRNA和TGFβ1/ Smad3途径抑制剂（SB431542）。肾脏组织学用于检测肾脏变化。采用实时荧光定量PCR和western blot分析VASH-1，SIRT1，HIF1α，TGFβ1，Smad3，血管内皮生长因子（VEGF），结缔组织生长因子（CTGF）和纤连蛋白（FN）的表达。用ELISA法检测大鼠组织和细胞培养上清液中肿瘤坏死因子-α（TNFα），TGFβ1，超氧化物歧化酶（SOD），过氧化氢酶（CAT），谷胱甘肽过氧化物酶（GSH-PX）和丙二醛（MDA）的表达水平。化学发光测定，而CCK-8检测细胞增殖。

结果： 糖尿病大鼠肾组织中VASH-1水平降低，高糖和高糖均降低。 Vash1 siRNA抑制RMC中VASH-1和SIRT1的表达，增加HIF1α，TGFβ1和Smad3的水平，从而上调氧化应激和纤维化因子，并异常增加细胞增殖活性（P<0.05）。但是，抑制SIRT1 /HIF1α信号通路只会降低TGFβ1和Smad3（P <0.05），而VASH-1保持不变（P > 0.05）。

结论： VASH-1在糖尿病大鼠肾组织中表达不足，并通过下游SIRT1 /HIF1α和TGFβ1/ Smad3信号通路调节DKD中氧化应激和纤维化的病理过程。