目的 研究并探讨枸杞多糖对原代培养新生大鼠海马神经元氧糖剥夺再灌注(oxygen-glucose deprivation/reperfusion,OGD/RP)损伤的保护作用。方法 以原代培养的新生大鼠海马神经元为研究对象建立氧糖剥夺再灌注损伤模型。MTT法测定神经细胞的存活率,分光光度计检测乳酸脱羟酶(LDH)漏出率、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)含量,激光共聚焦显微镜(CLSM)测定海马神经元细胞内Ca2+浓度和线粒体膜电位水平(MMP)。结果 与氧糖剥夺再灌注损伤组比较,枸杞多糖(10,20,40 mg·L-1)组可显著提高细胞存活率(P＜0.01),降低乳酸脱氢酶漏出率(P＜0.05,P＜0.01),升高超氧化物歧化酶和谷胱甘肽过氧化物酶活性(P＜0.01),降低丙二醛含量(P＜0.05,P＜0.01),抑制Ca2+浓度升高(P＜0.05,P＜0.01),枸杞多糖(20,40 mg·L-1)组能显著降低线粒体膜电位水平(P＜0.01)。结论 枸杞多糖对海马神经元氧糖剥夺再灌注损伤具有明显保护作用。
Abstract
To investigate the protective effects of Lycium barbarum polysaccharide(LBP) against injury from oxygen-glucose deprivation/reperfusion (OGD/RP)in primary cultured rat hippocampal neurons. METHODS Cultured hippocampal neurons were exposed to oxygen-glucose deprivation (OGD)for 2 h followed by a 24 h reperfusion. The MTT assay and the lactate dehydrogenase (LDH)release were used to evaluate the protective effects of LBP. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA)were determined by spectrophotometry using commercial kits. Mitochondrial membrane potential (MMP)and the intracellular free calcium concentration (i)in hippocampal neurons were measured using confocal laser scanning microscope (CLSM). RESULTS Treatment with LBP (10-40 mg稬-1)significantly attenuated neuronal damage and inhibited LDH release in a dose-dependent manner. Furthermore, LBP enhanced activities of SOD and GSH-Px but it decreased their MDA content, inhibited i elevation and decrease of MMP in ischemia-reperfusion treated hippocampal neurons. CONCLUSION These findings suggested that LBP may be a potential neuroprotective agent for cerebral ischemia-reperfusion injury.
关键词
枸杞多糖 /
海马神经元 /
氧糖剥夺 /
再灌注损伤 /
神经保护
{{custom_keyword}} /
Key words
Lycium barbarum polysaccharide(LBP) /
hippocampal neurons /
oxygen-glucose deprivation /
reperfusion /
neuroprotection
{{custom_keyword}} /
中图分类号：
R965
{{custom_clc.code}}
({{custom_clc.text}})
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] GENOVESE T, MAZZON E, PATERMITI I, et al. Neuroprotective effects of olprinone after cerebral ischemia/reperfusion injury in rats. Neuroscience Letters, 2011, 503(2):93-99. [2] RYOU M G, LIU R, REN M, et al. Pyruvate protects the brain against ischemia reperfusion injury by activating the erythropoietin signaling pathway. Stroke, 2012, 43(4):1101-1107. [3] DU G, LIU L, FANG J. Experimental study on the enhancement of murine splenic lymphocyte proliferation by Lycium barbarum glycopeptide. J Huazhong Univ Sci Technolog Med Sci(华中科技大学医学院学报), 2004, 24(5):518-520, 527. [4] GAN L, ZHANG S H, LIU Q, et al. A polysaccharide-protein complex from Lycium barbarum upregulates cytokine expression in human peripheral blood mono nucle-ar cells. Eur J Pharmacol, 2003, 471(3):217-222. [5] LI X M, MA Y L, LIU X J. Effect of the Lycium barbarum polysaccharides on age-related oxidative stress in aged mice. J Ethnopharmacol, 2007, 111(3):504-511. [6] YU MS, LAI C S, HO Y S, et al. Characterization of the effects of anti-aging medicine Fructus lycii on beta-amyloid peptide neurotoxicity. Int J Mol Med, 2007, 20(2):261-268. [7] HA K T, YOON S J, CHOI D Y, et al. Protective effect of ycium chinense fruit on carbon tetrachloride-induced hepatotoxicity. J Ethnopharmacol, 2005, 96(3):529-535. [8] LUO Q, CAI Y, YAN J,et al. Hypoglycemic and hypolipidemic ffects and antioxidant activity of fruit extracts from Lycium barbarum. Life Sci, 2004, 76(2):137-149. [9] GONG H Y, LI J Y, LIANG L, et al. Protective effect of polysaccharides of Lycium barbarum on neuron injured by hypoxia . Acta Acad Med CPAPF(武警医学院学报), 2009, 18(12):1002-1004. ZHAO J, WU Y, SUN M, et al. Protective effects and mechanisms of OSR on primary cultured hippocampus neurons subjected to anoxic injury in neonatal rat . China J Chin Mater Med(中国中药杂志), 2012, 37(1):94-98. REERSM, SMITH T W, CHEN L B. 1991. J-aggregate formation of a carbocyanine as a quantitative fluorescent indicator of membrane potential. Biochemistry, 30 (18):4480-4486. ZHU X J, SHI Y, PENG J, et al. The effects of BAFF and BAFF-R-Fc fusion protein in immune thrombocytopenia. Blood, 2009, 114(26):5362-5367. PASTIL, CARMIGNOTO G, POZZAN T, et al. Cellularcalcium handling in brain slices from calbindin D28k-defcient mice. Neuroreport, 1999, 10(1):2367-2372. REN Z C, GUO J X, YANG Y C, et al. Expression of cyclooxygenase in the cerebral cortex and protective effect of tetramethylpyrazine after cerebral ischemic-reperfusion. Chin J Anatomy, 2008, 31(6):831-835. SCHALLER B, GRAF R. Cerebral ischemia and reperfusion: The pathophysiologic concept as a basis for clinical therapy. J Cereb Blood Flow Metab, 2004, 24(4):351-371. RAMIREZ M R, MURARO F, ZYLBERSZTEJN D S, et al. Neonatal hypoxia-ischemia reduces ganglioside, phospholipid and cholesterol contents in the rat hippocampus. Neurosci Res, 2003, 46(3):339-347. MUNNS S E, MELONI B P, KNUCKEY N W, et al. Primary cortical neuronal cultures reduce cellular energy utilization during anoxic energy deprivation. J Neurochem, 2003, 87(3):764-772. REN Y H, LIU X N, ZHOU B Y, et al. Neuroprotective effect of paeoniflorin on anoxically cultured rat hippoeampal neurons. J Four Mil Med Univ(第四军医大学学报), 2006, 27(16):1480-1482. ZHANG X D, LI W H, WANG Q, et al. Protective effects of ginsenoside Rg1, Re on cultured hippoc ampal and cortex neurons injury induced by β-amyloid protein . Acad J Sec Mil Med Univ(第二军医大学学报), 2002, 21(10):941-943. WANG Q, GONG Q, SHI J, et al. Neuroprotective effects of Dendrobium alkaloids on rat cortical neurons injured by oxygen-glucose deprivation and reperfusion. Phytomedicine, 2000, 17(2):108-115. DAVID J M, STEN O. The role of calcium in the regulation of apoptosis. J Leukoc Biol, 1996, 59(1):775-783. AABDALLAH D M, EID N I. Possible neuroprotective effects of lecithin and alpha-tocopherol alone or in combination against ischemia/reperfusion insult in rat brain. J Biochem Mol Toxic, 2004, 18(5):273-278. NITA D A, NITA V, SPULBER S, et al. Oxidative damage following cerebral ischemia depends on reperfusion-a biochemical study in rat. J Cel Mol Med, 2001, 5(2):163-170.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
基金
国家重点基础研究发展计划(973计划)(2010CB735603)
{{custom_fund}}
PDF(814 KB)
