紫檀芪激活血红素加氧酶-1减轻小鼠脑缺血再灌注后线粒体氧化损伤
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摘要
目的:探索紫檀芪(pterostilbene,PTE)对小鼠脑缺血再灌注(IR)后线粒体氧化损伤的作用及可能机制。方法:通过双侧颈总动脉阻断法建立小鼠脑IR模型,腹腔注射PTE或Zn PP(血红素加氧酶-1抑制剂),动物随机均分为IR组、PTE+IR组、PTE+Zn PP+IR组、Zn PP+IR组,并且PTE有2.5 mg/kg、5 mg/kg、10 mg/kg三个剂量组。然后,进行神经功能评分;通过干湿比重法测定脑水肿;火焰光度法测定Na~+含量;Neu N和TUNEL法测定细胞存活和凋亡;通过线粒体膜电位(MMP)、线粒体活性氧(ROS)产物和线粒体复合物I和IV活性测定来检测线粒体的氧化应激损伤;通过Western Blot检测血红素加氧酶-1(HO-1)、NADPH醌氧化还原酶-1(NQO1)、谷胱甘肽S-转移酶(GST)、线粒体和胞浆细胞色素c蛋白的表达。结果:PTE可以提高小鼠的神经功能评分、减轻脑水肿和降低脑Na~+含量。PTE上调HO-1、NQO1和GST的表达,提高MMP、线粒体复合体I/IV活性和线粒体细胞色素c水平,减少线粒体ROS产物和降低胞浆细胞色素c水平,并且有一定的剂量依赖性。但是,PTE的这些作用可以被HO-1的抑制剂Zn PP逆转。结论:在脑IR模型小鼠,PTE通过激活HO-1信号减轻线粒体氧化应激损伤和细胞死亡,从而发挥脑保护作用。
Objective: To explore the effects of pterostilbene(PTE) on mitochondrial oxidative damage in mice cerebral ischemia reperfusion( IR) model and the possible mechanisms. Methods: IR injury was induced by bilateral common carotid arteries occlusion in mice using non-traumatic aneurysm clips. The mice received PTE or Zn PP,the inhibitor of heme oxygenase-1( HO-1) by intraperitoneal injection. The mice were randomly divided into four groups: IR; PTE +IR; PTE + Zn PP + IR; Zn PP + IR. The dose of PTE used was 2. 5 mg/kg,5 mg/kg and 10 mg/kg. The neurological scores were assessed and the brain edema was detected by wet-dried weight method. The sodium ion level in the brain was measured by flame photometry. The neuronal survival and the apoptotic ratio in IR-injured brains were detected by Neu N and TUNEL assay. The mitochondrial oxidative stress injury was evaluated through the measurements of mitochondrialmembrane potential( MMP),reactive oxygen species( ROS) production,mitochondrial complex I and IV activity. The protein expressions of HO-1,NADPH quinone oxidoreductase 1( NQO1),glutathione s-transferases( GST),mitochondrial and cytosolic cytochrome c was detected by Western Blot. Results: PTE improved neurological scores of mice,lowered brain edema and decreased the sodium ion content of brain. It up-regulated HO-1,NQO1 and GST expression.Moreover,PTE resulted in a well-preserved mitochondrial MMP,mitochondria complex I/IV activity,mitochondrial cytochrome c level while it reduced mitochondrial ROS production and cytosolic cytochrome c level. However,the dosedependent PTE-elevated mitochondrial function was reversed by Zn PP. Conclusion: In mice cerebral IR model,PTE plays a role in the brain protection by reducing mitochondrial oxidative damage and cell death through the activation of HO-1 signal.
Objective: To explore the effects of pterostilbene(PTE) on mitochondrial oxidative damage in mice cerebral ischemia reperfusion( IR) model and the possible mechanisms. Methods: IR injury was induced by bilateral common carotid arteries occlusion in mice using non-traumatic aneurysm clips. The mice received PTE or Zn PP,the inhibitor of heme oxygenase-1( HO-1) by intraperitoneal injection. The mice were randomly divided into four groups: IR; PTE +IR; PTE + Zn PP + IR; Zn PP + IR. The dose of PTE used was 2. 5 mg/kg,5 mg/kg and 10 mg/kg. The neurological scores were assessed and the brain edema was detected by wet-dried weight method. The sodium ion level in the brain was measured by flame photometry. The neuronal survival and the apoptotic ratio in IR-injured brains were detected by Neu N and TUNEL assay. The mitochondrial oxidative stress injury was evaluated through the measurements of mitochondrialmembrane potential( MMP),reactive oxygen species( ROS) production,mitochondrial complex I and IV activity. The protein expressions of HO-1,NADPH quinone oxidoreductase 1( NQO1),glutathione s-transferases( GST),mitochondrial and cytosolic cytochrome c was detected by Western Blot. Results: PTE improved neurological scores of mice,lowered brain edema and decreased the sodium ion content of brain. It up-regulated HO-1,NQO1 and GST expression.Moreover,PTE resulted in a well-preserved mitochondrial MMP,mitochondria complex I/IV activity,mitochondrial cytochrome c level while it reduced mitochondrial ROS production and cytosolic cytochrome c level. However,the dosedependent PTE-elevated mitochondrial function was reversed by Zn PP. Conclusion: In mice cerebral IR model,PTE plays a role in the brain protection by reducing mitochondrial oxidative damage and cell death through the activation of HO-1 signal.
引文
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[16]Moreira TJ,Cebere A,Cebers G,et al.Reduced HO-1 protein expression is associated with more severe neurodegeneration after transient ischemia induced by cortical compression in diabetic Goto-Kakizaki rats[J].J Cereb Blood Flow Metab,2007,27:1710-1723.
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[18]Chiou YS,Tsai ML,Nagabhushanam K,et al.Pterostilbene is more potent than resveratrol in preventing azoxymethane(AOM)-induced colon tumorigenesis via activation of the NF-E2-related factor2(Nrf2)-mediated antioxidant signaling pathway[J].J Agric Food Chem,2011,59:2725-2733.
[19]Zhang L,Zhou G,Song W,et al.Pterostilbene protects vascular endothelial cells against oxidized low-density lipoprotein-induced apoptosis in vitro and in vivo[J].Apoptosis,2012,17:25-36.
[20]Cui Y,Zhang H,Ji M,et al.Hydrogen-rich saline attenuates neuronal ischemia-reperfusion injury by protecting mitochondrial function in rats[J].J Surg Res,2014,192:564-572.
[2]Li N,Ma Z,Li M,et al.Natural potential therapeutic agents of neurodegenerative diseases from the traditional herbal medicine Chinese dragon’s blood[J].J Ethnopharmacol,2014,52:508-521.
[3]Hou Y,Xie G,Miao F,et al.Pterostilbene attenuates lipopolysaccharide-induced learning and memory impairment possibly via inhibiting microglia activation and protecting neuronal injury in mice[J].Prog Neuropsychopharmacol Biol Psychiatry,2014,54:92-102.
[4]Yang Y,Duan W,Li Y,et al.New role of silent information regulator 1 in cerebral ischemia[J].Neurobiol Aging,2013,34:2879-2888.
[5]Wang YF,Gu YT,Qin GH,et al.Curcumin ameliorates the permeability of the blood-brain barrier during hypoxia by upregulating heme oxygenase-1 expression in brain microvascular endothelial cells[J].J Mol Neurosci,2013,51:344-351.
[6]Shin JH,Kim SW,Jin Y,et al.Ethyl pyruvatemediated Nrf2 activation and hemeoxygenase 1 induction in astrocytes confer protective effects via autocrine and paracrine mechanisms[J].Neurochem Int,2012,61:89-99.
[7]Bhakkiyalakshmi E,Shalini D,Sekar TV,et al.Therapeutic potential of pterostilbene against pancreatic beta-cell apoptosis mediated through Nrf2[J].Br J Pharmacol,2014,171:1747-1757.
[8]Colin-Gonzalez AL,Orozco-Ibarra M,Chanez-Cardenas ME,et al.Heme oxygenase-1(HO-1)upregulation delays morphological and oxidative damage induced in an excitotoxic/pro-oxidant model in the rat striatum[J].Neuroscience,2013,231:91-101.
[9]Zhang HP,Sun YY,Chen XM,et al.The neuroprotective effects of isoflurane preconditioning in a murine transient global cerebral ischemiareperfusion model:the role of the Notch signaling pathway[J].Neuromolecular Med,2014,16:191-204.
[10]Hou Y,Xie G,Miao F,et al.Pterostilbene attenuates lipopolysaccharide-induced learning and memory impairment possibly via inhibiting microglia activation and protecting neuronal injury in mice[J].Prog Neuropsychopharmacol Biol Psychiatry,2014,54:92-102.
[11]Hwang JH,Kim YH,Noh JR,et al.The protective role of NAD(P)H:quinone oxidoreductase 1 on acetaminophen-induced liver injury is associated with prevention of adenosine triphosphate depletion and improvement of mitochondrial dysfunction[J].Arch Toxicol,2015,2159-2166.
[12]Ulziikhishig E,Lee KK,Hossain QS,et al.Inhibition of mitochondrial membrane bound-glutathione transferase by mitochondrial permeability transition inhibitors including cyclosporin A[J].Life Sci,2010,86:726-732.
[13]Chen G,Fang Q,Zhang J,et al.Role of the Nrf2-ARE pathway in early brain injury after experimental subarachnoid hemorrhage[J].J Neurosci Res,2011,89:515-523.
[14]Takizawa S,Hirabayashi H,Matsushima K,et al.Induction of heme oxygenase protein protects neurons in cortex and striatum,but not in hippocampus,against transient forebrain ischemia[J].J Cereb Blood Flow Metab,1998,18:559-569.
[15]Panahian N,Yoshiura M,Maines MD.Overexpression of heme oxygenase-1 is neuroprotective in a model of permanent middle cerebral artery occlusion in transgenic mice[J].J Neurochem,1999,72:1187-1203.
[16]Moreira TJ,Cebere A,Cebers G,et al.Reduced HO-1 protein expression is associated with more severe neurodegeneration after transient ischemia induced by cortical compression in diabetic Goto-Kakizaki rats[J].J Cereb Blood Flow Metab,2007,27:1710-1723.
[17]Bhakkiyalakshmi E,Shalini D,Sekar TV,et al.Therapeutic potential of pterostilbene against pancreatic beta-cell apoptosis mediated through Nrf2[J].Br J Pharmacol,2014,171:1747-1757.
[18]Chiou YS,Tsai ML,Nagabhushanam K,et al.Pterostilbene is more potent than resveratrol in preventing azoxymethane(AOM)-induced colon tumorigenesis via activation of the NF-E2-related factor2(Nrf2)-mediated antioxidant signaling pathway[J].J Agric Food Chem,2011,59:2725-2733.
[19]Zhang L,Zhou G,Song W,et al.Pterostilbene protects vascular endothelial cells against oxidized low-density lipoprotein-induced apoptosis in vitro and in vivo[J].Apoptosis,2012,17:25-36.
[20]Cui Y,Zhang H,Ji M,et al.Hydrogen-rich saline attenuates neuronal ischemia-reperfusion injury by protecting mitochondrial function in rats[J].J Surg Res,2014,192:564-572.