Liu Dajun, Shang Huiping, Liu Ying
Department of Nephrology, Shengjing Affiliated Hospital of China Medical University, Shenyang 110036, China.
Int J Mol Sci. 2016 Jul 12;17(7):1051. doi: 10.3390/ijms17071051.
Stanniocalcin-1 (STC-1) protects against renal ischemia-reperfusion injury (RIRI). However, the molecular mechanisms remain widely unknown. STC-1 inhibits reactive oxygen species (ROS), whereas most ROS-mediated pathways are associated with ischemic injury. Therefore, to explore the mechanism, the effects of STC-1 on ROS-medicated pathways were studied. Non-traumatic vascular clamps were used to establish RIRI mouse models. The serum levels of STC-1, interleukin-6 (IL-6), interferon (IFN) γ, P53, and capase-3 were measured by ELISA kits. Superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by fluorescence spectrofluorometer. All these molecules changed significantly in a RIRI model mouse when compared with those in a sham control. Kidney cells were isolated from sham and model mice. STC-1 was overexpressed or knockout in these kidney cells. The molecules in ROS-medicated pathways were measured by real-time quantitative PCR and Western blot. The results showed that STC-1 is an effective ROS scavenger. The serum levels of STC-1, MDA and SOD activity were increased while the serum levels of IL-6, iIFN-γ, P53, and capase-3 were decreased in a model group when compared with a sham control (p < 0.05). Furthermore, the levels of STC-1,p53, phosphorylated mitogen-activated protein kinase kinase (p-MEKK-1), c-Jun N-terminal kinase (p-JNK), extracellular signal-regulated kinase (p-ERK), IkB kinase (p-IKK), nuclear factor (NF) κB, apoptosis signal-regulating kinase 1 (ASK-1) and caspase-3 changed significantly in kidney cells isolated from a RIRI model when compared to those isolated from a sham control (p < 0.05). Meanwhile, STC-1 overexpression or silence caused significant changes of the levels of these ROS-mediated molecules. Therefore, STC-1 maybe improve anti-inflammation, anti-oxidant and anti-apoptosis activities by affecting ROS-mediated pathways, especially the phospho-modifications of the respective proteins, resulting in the increase of SOD and reduce of capase-3, p53, IL-6 and IFN-γ.
1,25-二羟基维生素D3（STC-1）可保护肾脏免受缺血再灌注损伤（RIRI）。然而，其分子机制仍广泛未知。STC-1可抑制活性氧（ROS），而大多数ROS介导的途径与缺血性损伤有关。因此，为探究其机制，研究了STC-1对ROS介导途径的影响。采用无创血管夹建立RIRI小鼠模型。用ELISA试剂盒检测血清中STC-1、白细胞介素-6（IL-6）、干扰素（IFN）γ、P53和半胱天冬酶-3的水平。用荧光分光光度计检测超氧化物歧化酶（SOD）和丙二醛（MDA）。与假手术对照组相比，RIRI模型小鼠的所有这些分子均发生了显著变化。从假手术和模型小鼠中分离肾细胞。在这些肾细胞中过表达或敲除STC-1。通过实时定量PCR和蛋白质印迹法检测ROS介导途径中的分子。结果表明，STC-1是一种有效的ROS清除剂。与假手术对照组相比，模型组血清中STC-1、MDA水平和SOD活性升高，而血清中IL-6、IFN-γ、P53和半胱天冬酶-3水平降低（p<0.05）。此外，与从假手术对照组分离的肾细胞相比，从RIRI模型分离的肾细胞中STC-1、p53、磷酸化丝裂原活化蛋白激酶激酶（p-MEKK-1）、c-Jun氨基末端激酶（p-JNK）、细胞外信号调节激酶（p-ERK）、IkB激酶（p-IKK）、核因子（NF）κB、凋亡信号调节激酶1（ASK-1）和半胱天冬酶-3的水平发生了显著变化（p<0.05）。同时，STC-1的过表达或沉默导致这些ROS介导分子水平的显著变化。因此，STC-1可能通过影响ROS介导的途径，尤其是各蛋白的磷酸化修饰，提高抗炎、抗氧化和抗凋亡活性，从而导致SOD增加，半胱天冬酶-3、p53、IL-6和IFN-γ减少。
