Reed Dustin K, Hall Samuel, Arany Istvan
Department of Pediatrics, University of Mississippi Medical Center, Jackson, MS, USA.
J Physiol Biochem. 2015 Mar;71(1):1-7. doi: 10.1007/s13105-014-0372-x. Epub 2014 Dec 4.
Smoking and obesity increases renal oxidative stress via nicotine (NIC) or free fatty acid such as oleic acid (OA) but decreases levels of the vitamin E-derivative α-tocopherol (TOC), which has shown to stimulate the antioxidant system such as heme oxygenase-1 (HO-1). Hence, we hypothesized that supplementation of TOC may protect renal proximal tubules from NIC- or OA-mediated oxidative stress by upregulating the HO-1 gene. NIC- or OA-dependent production of reactive oxygen species (ROS) was determined in the presence or absence of various pharmacologic or genetic inhibitors that modulate HO-1 activation and enhancer elements in the HO-1 promoter such as the antioxidant response element (ARE) and the cAMP-response element (CRE) in renal proximal tubule cells (NRK52E). Activity of the HO-1 promoter, the ARE and the CRE was determined in luciferase assays. We found that pre- or posttreatment with TOC attenuated NIC- or OA-dependent ROS production that required HO-1 activation. TOC activated the HO-1 promoter via the CRE but not the ARE enhancer through the extracellular signal-regulated kinase (ERK) and protein kinase A (PKA). Consequently, inhibitors of ERK, PKA, or CRE activation mitigated beneficial effects of TOC on NIC- or OA-mediated ROS production. Hence, vitamin E supplementation-via induction of the cytoprotective HO-1-may help to reduce renal oxidative stress imposed by smoking or obesity.
吸烟和肥胖会通过尼古丁(NIC)或游离脂肪酸(如油酸,OA)增加肾脏氧化应激,但会降低维生素E衍生物α-生育酚(TOC)的水平,而α-生育酚已被证明可刺激抗氧化系统,如血红素加氧酶-1(HO-1)。因此,我们推测补充TOC可能通过上调HO-1基因来保护肾近端小管免受NIC或OA介导的氧化应激。在存在或不存在各种调节HO-1激活和HO-1启动子中增强子元件(如抗氧化反应元件(ARE)和cAMP反应元件(CRE))的药理或基因抑制剂的情况下,测定肾近端小管细胞(NRK52E)中NIC或OA依赖性活性氧(ROS)的产生。通过荧光素酶测定法测定HO-1启动子、ARE和CRE的活性。我们发现,TOC预处理或后处理可减弱需要HO-1激活的NIC或OA依赖性ROS产生。TOC通过细胞外信号调节激酶(ERK)和蛋白激酶A(PKA)激活HO-1启动子,但不通过ARE增强子激活。因此,ERK、PKA或CRE激活的抑制剂减轻了TOC对NIC或OA介导的ROS产生的有益作用。因此,补充维生素E——通过诱导细胞保护性HO-1——可能有助于减少吸烟或肥胖引起的肾脏氧化应激。