长白山核桃五肽对过氧化氢诱导PC12细胞的保护作用及机理研究

摘要/Abstract

摘要： 本文以长白山核桃料五肽Leu-Pro-Leu-Leu-Arg(LPLLR)为研究对象，利用细胞试验和NF-κB/iNOS/NO信号通路探讨对过氧化氢诱导PC12细胞保护作用的机理。结果显示，LPLLR对PC12细胞无毒副作用，能显著提高受氧化损伤PC12细胞的存活率，降低细胞中活性氧簇（reactive oxygen species，ROS）（P＜0.01）和丙二醛 (Malondialdehyde，MDA) （P＜0.05）含量，提高过氧化物歧化酶（Superoxide Dismutase，SOD）（P＜0.01）、过氧化氢酶（Catalase，CAT）（P＜0.05）、谷胱甘肽过氧化物酶(Glutathione peroxidase，GSH-Px)（P＜0.01）活性和乙酰胆碱（Acetylcholine，ACh）（P＜0.05）含量。通过ELISA法和Western blotting检测NO分泌量和SOD2、NF-κB p65、iNOS蛋白表达水平，LPLLR能降低NO (P＜0.01)分泌量和上调 SOD2蛋白表达水平（P＜0.01），下调NF-κB p65（P＜0.05）和iNOS（P＜0.05）蛋白表达水平。表明LPLLR对氧化应激损伤的PC12细胞具有一定的保护作用。

关键词: 核桃水解肽, 过氧化氢, PC12细胞, 氧化应激

Abstract: The mechanism of the pentapeptide Leu-Pro-Leu-Leu-Arg (LPLLR) which was from Changbai Mountain Walnut on the protection of PC12 cells induced by hydrogen peroxide was investigated by cell experiments and NF-κB/iNOS/NO signaling pathway. The results showed that LPLLR had no cytotoxicity and could significantly increase the survival rate of PC12 cells which were damaged by oxidative injury. LPLLR could decrease the production of ROS (P＜0.01) and MDA (P＜0.05), and increase the production of SOD（P＜0.01), CAT (P＜0.05), GSH-Px（P＜0.01）and ACh (P＜0.05) on PC12 cells induced by hydrogen peroxide. The production of NO and the protein expression of SOD2, NF-κB p65, iNOS were determined by ELISA and Western blotting. The results confirmed that LPLLR could up-regulate the expression of SOD2 (P＜0.05) and down-regulate the expression of NF-κB p65 (P＜0.05) and iNOS (P＜0.05), decreasing the secretion of NO (P＜0.01). It indicates that LPLLR has a protective effect on PC12 cells injured by hydrogen peroxide.

Key words: walnut peptides, hydrogen peroxide, PC12 cells, oxidative stress

中图分类号:

TS209

郭勇秦汉雄魏贞方丽闵伟红. 长白山核桃五肽对过氧化氢诱导PC12细胞的保护作用及机理研究[J]. 食品科学, 0, (): 0-0.

参考文献

[1]崔玲玉, 闵伟红, 刘威, 等.长白山核桃楸分离蛋白缓解小鼠体力疲劳的作用[J].食品科学, 2016, 37(9):203-207 [2]刘威, 闵伟红, 刘春雷, 等.核桃清蛋白抗氧化肽的制备及其活性研究[J].食品研究与开发, 2017, 38(17):40-44 [3]赵谋明, 任娇艳, 等.食源性生物活性肽结构特征与生理活性的研究现状与趋势[J].中国食品学报, 2011, 11(9):69-81 [4]ELLERT-MIKLASZEWSKA A, POLESZAK K, KAMINSKA B.Short peptides interfering with signaling pathways as new therapeutic tools for cancer treatment[J].Future Medicinal Chemistry, 2017, 9(2):199-221 [5]JIANG F, GUAN H, LIU D, et al.Flavonoids from sea buckthorn inhibit the lipopolysaccharide-induced inflammatory response in RAW2647 macrophages through the MAPK and NF-κB pathways[J].Food & Function, 2017, 8(3):1313-1322 [6] LIU C, FANG L, MIN W, et al.Exploration of the molecular interactions between angiotensin-I-converting enzyme (ACE) and the inhibitory peptides derived from hazelnut (Corylus heterophylla Fisch.)[J]. Food Chemistry, 2018:471-480. DOI: 10.1016/j.foodchem.2017.10.095.[J].Food Chemistry, 2018, 245(1):471-480 [7]KIM Y, KIM I, CHOI J, et al.The anti-obesity effects of a tuna peptide on 3T3-L1 adipocytes are mediated by the inhibition of the expression of lipogenic and adipogenic genes and by the activation of the Wntβ-catenin signaling pathway[J].International Journal of Molecular Medicine, 2015, 36(2):327-334 [8] CHEN H, ZHAO M, LIN L, et al.Identification of antioxidative peptides from defatted walnut meal hydrolysate with potential for improving learning and memory.[J]. Food Research International, 2015, 78:216-223. DOI: 10.1016/j.foodres.2015.10.008.[J]. Food Research International, 2015, 78(1):216-223 [9] GIUDETTI A M, SALZET M, CASSANO T.Oxidative Stress in Aging Brain: Nutritional and Pharmacological Interventions for Neurodegenerative Disorders[J]. Oxidative Medicine & Cellular Longevity, 2018, 2018:1-2. DOI: 10.1155/2018/3416028.[J].Oxidative Medicine & Cellular Longevity, 2018, 2018(1):1-2 [10]STAUFFER J, PANDA B, ILMONEN P.Telomere length,sibling competition and development of antioxidant defense in wild house mice[J].Mechanisms of Ageing & Development, 2017, 07(169):45-52 [11]ALI T, BADSHAH H, KIM T H, et al.Melatonin attenuates D-galactose-induced memory impairment,neuroinflammation and neurodegeneration via RAGENF-κBJNK signaling pathway in aging mouse model[J].Journal of Pineal Research, 2015, 58(1):71-85 [12]POLCE S A, BURKE C, FRAN?A L M, et al.Ellagic Acid Alleviates Hepatic Oxidative Stress and Insulin Resistance in Diabetic Female Rats[J].Nutrients, 2018, 10(5):531-546 [13] GIRALDO E, LLORET A, FUCHSBERGER T, et al.Aβ and tau toxicities in Alzheimer’s are linked via oxidative stress-induced p38 activation: Protective role of vitamin E[J]. Redox Biology, 2014, 2:873-877. DOI: 10.1016/j.redox.2014.03.002.[J].Redox Biology, 2014, 2(1):873-877 [14]REYNOLDS G D.Aldosterone-induced oxidative stress and inflammation in the brain are mediated by the endothelial cell mineralocorticoid receptor[J].Brain Research, 2016, 1637(88):146-153 [15]BRIEGER K, SCHIAVONE S, JR M F, et al.Reactive oxygen species: from health to disease[J].Swiss Medical Weekly, 2012, 142(9842):13659-13673 [16]LI Y, ZHANG W, CHEN C, et al.Inotodiol protects PC12 cells against injury induced by oxygen and glucose deprivationrestoration through inhibiting oxidative stress and apoptosis[J].Journal of Applied Biomedicine, 2017, 16(2):126-132 [17]CHO C H, KIM E A, KIM J, et al.N-Adamantyl-4-methylthiazol-2-amine suppresses amyloid β-induced neuronal oxidative damage in cortical neurons[J].Free Radical Research, 2016, 50(6):678-690 [18]FEUERSTEIN D, BACKES H, GRAMER M, et al.Regulation of cerebral metabolism during cortical spreading depression[J].Journal of Cerebral Blood Flow & Metabolism Official Journal of the International Society of Cerebral Blood Flow & Metabolism, 2015, 36(11):1965-1977 [19] 王祖浩, 郭勇, 周美含, 等.榛仁五肽对HUVEC细胞氧化应激损伤的保护作用[J]. 现代食品科技, 2018(3). DOI：10.13982/j.mfst.1673-9078.2018.03.001.[J].现代食品科技, 2018, 3(1):1-7 [20]KAWASHIMA K, ITOH H, MIYOSHI M, et al.Antioxidant properties of branched-chain amino acid derivatives[J].Chemical & Pharmaceutical Bulletin, 1979, 27(8):1912-1916 [21]CHEISON S C, WANG Z, XU S Y.Preparation of whey protein hydrolysates using a single- and two-stage enzymatic membrane reactor and their immunological and antioxidant properties: characterization by multivariate data analysis[J].Journal of Agricultural & Food Chemistry, 2007, 55(10):3896-3904 [22]PADURARIU M, CIOBICA A, HRITCU L, et al.Changes of some oxidative stress markers in the serum of patients with mild cognitive impairment and Alzheimer's disease[J].Neuroscience Letters, 2010, 469(1):6-10 [23]ZHU Y, HOELL P, AHLEMEYER B, et al.Implication of PTEN in production of reactive oxygen species and neuronal death in in vitro models of stroke and Parkinson's disease[J].Neurochemistry International, 2007, 50(3):507-516 [24]FERRER I, PEREZ E, DALFó E, et al.Abnormal levels of prohibitin and ATP synthase in the substantia nigra and frontal cortex in Parkinson's disease[J].Neuroscience Letters, 2007, 415(3):205-209 [25] JIA J, ZHANG L, SHI X, et al.SOD2 Mediates Amifostine-Induced Protection against Glutamate in PC12 Cells[J]. Oxidative Medicine and Cellular Longevity, 2016(6):1-11. DOI: 10.1155/2016/4202437.[J].Oxidative Medicine and Cellular Longevity, 2016, 6(1):1-11 [26]HERSCOVITCH M, COMB W, ENNIS T, et al.Intermolecular disulfide bond formation in the NEMO dimer requires Cys54 and Cys347[J].Biochemical & Biophysical Research Communications, 2008, 367(1):103-108