Please wait a minute...
浙江大学学报（医学版）
浙江大学学报（医学版）  2015, Vol. 44 Issue (1): 24-29    DOI: 10.3785/j.issn.1008-9292.2015.01.004
专题报道     
葡萄籽原花青素对氯化钴诱导的视网膜神经节细胞RGC-5缺氧损伤的保护作用
林卡娜1,2, 林美丽1, 魏尔清2
1. 浙江大学医学院附属第二医院药剂科, 浙江 杭州 310009;
2. 浙江大学医学院药理学系, 浙江 杭州 310058
Protective effect of grape seed proanthocyanidin on cultured RGC-5 cells against CoCl2-induced hypoxic injury
LIN Ka-na1,2, LIN Mei-li1, WEI Er-qing2
1. Department of Pharmacy, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China;
2. Department of Pharmacology, Zhejiang University School of Medicine, Hangzhou 310058, China
全文: PDF(1827 KB)  
摘要: 

目的:探讨体外葡萄籽原花青素对氯化钴致视网膜神经节细胞RGC-5缺氧损伤模型的保护作用.方法:采用氯化钴(400 μmol/L)诱导RGC-5细胞化学缺氧损伤,观察葡萄籽原花青素(40 μg/mL)对缺氧的抗损伤作用.以MTT法检测细胞存活率;Hoechst 33342荧光染色法观察细胞凋亡情况;以H2DCFDA荧光法检测细胞内活性氧的变化;实时定量PCR检测细胞Bcl-2、半胱酰胺天冬氨酸蛋白酶(caspase) 9和caspase 3 mRNA水平.结果:与缺氧对照组比较,葡萄籽原花青素预处理可提高细胞存活率(P <0.001)、减少细胞凋亡(P <0.001),并减少细胞内的活性氧(P <0.001);葡萄籽原花青素还显著升高Bcl-2基因的表达(P <0.001),显著降低caspase 9和caspase 3基因的表达(P <0.001).结论:葡萄籽原花青素对氯化钴诱导的视网膜神经节RGC-5细胞缺氧损伤起保护作用,其保护作用可能与细胞内活性氧减少、Bcl-2基因表达上调及casepase 9和casepase 3基因表达下调有关.
关键词 ： 葡萄/化学,  原花青素类/药理学,  钴/药理学,  视网膜神经节细胞/药物作用,  缺氧/治疗    
Abstract：

Objective: To investigate the protective effects of grape seed proanthocyanidin extracts (GSPE) against CoCl2-induced hypoxic injury in cultured RGC-5 cells. Methods: CoCl2 (400 μmol/L) was used to induce hypoxic injury in cultured RGC-5 cells; the cells were pretreated with 0, 100, 200, 400 and 800μmol/L GSPE for 24h. The cell viability was assayed by MTT; the apoptosis was detected by Hoechst 33342 staining; the intracellular reactive oxygen species (ROS) was measured by H2DCFDA oxidative reaction. The mRNA expression of Bcl-2, caspase 9 and caspase 3 was determined by real-time PCR. Results: Compared to hypoxic control group, pretreatment with GSPE significantly increased viability of RGC-5 cells (P <0.001), reduced cell apoptosis (P <0.001) and intracellular ROS (P <0.001). In addition, GSPE significantly increased the mRNA expression of Bcl-2 (P <0.001) and decreased mRNA expression of caspase 9 (P <0.001) and caspase 3 (P <0.001) compared to hypoxic control group. Conclusion: GSPE may have a protective effect against CoCl2-induced hypoxic injury in cultured RGC-5 cells. The decrease of intercellular ROS, up-regulation of Bcl-2 and down-regulation of caspase 9 and caspase 3 may be involved in the mechanism of the protective effect of GSPE.
Key words： VITIS VINIFERA/chemistry    Proanthocyanidins/pharmacology    Cobalt/pharmacology    Retinal ganglion cells/drug effects    Anoxia/therapy
收稿日期: 2014-08-20
CLC:  R96  
基金资助:

浙江省教育厅科学研究项目(Y201328427);浙江省中西医结合学会科研基金(2012LY013).
通讯作者: 魏尔清(1952-),男,博士,教授,博士生导师,从事神经药理学研究;E-mail: weieq2006@zju.edu.cn     E-mail: weieq2006@zju.edu.cn
作者简介: 林卡娜(1986-),女,硕士,初级药师,从事神经药理学研究;E-mail: linkana@163.com
服务　　
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章　　
林卡娜
林美丽
魏尔清

引用本文:

林卡娜, 林美丽, 魏尔清. 葡萄籽原花青素对氯化钴诱导的视网膜神经节细胞RGC-5缺氧损伤的保护作用[J]. 浙江大学学报（医学版）, 2015, 44(1): 24-29.
LIN Ka-na, LIN Mei-li, WEI Er-qing. Protective effect of grape seed proanthocyanidin on cultured RGC-5 cells against CoCl2-induced hypoxic injury. Journal of ZheJiang University(Medical Science), 2015, 44(1): 24-29.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.1008-9292.2015.01.004      或      http://www.zjujournals.com/xueshu/med/CN/Y2015/V44/I1/24

[1] SERNAGOR E, EGLEN S J, WONG R O. Development of retinal ganglion cell structure and function [J]. Prog Retin Eye Res, 2001, 20(2): 139-174.
[2] KRISHNAMOORTHY R R,AGARWAL P, PRASANNA G, et al. Characterization of a transformed rat retinal ganglion cell line [J]. Brain Res Mol Brain Res, 2001, 86(1-2):1-12.
[3] BAGCHI D, BAGCHI M, STOHS S J, et al. Free radicals and grape seed proanthocyanidin extract: importance in human health and disease prevention [J]. Toxicology, 2000, 148(2-3): 187-197.
[4] COS P,DE BRUYNE T, HERMANS N, et al. Proanthocyanidins in health care: current and new trends [J]. Curr Med Chem, 2004, 11(10): 1345-1359.
[5] BAGCHI DGARG A, KROHN R L, et al. Oxygen free radical scavenging abilities of vitamins C and E, and a grape seed proanthocyanidin extract in vitro [J]. Res Commun Mol Pathol Pharmacol, 1997, 95(2): 179-189.
[6] 黄晓瑾, 毛峻琴. 葡萄籽油原花青素抗大鼠缺血再灌注损伤的研究[J]. 中国药师, 2005,8(7):541-542. HUANG Xiao-jin,MAO Jun-qin. Protective effect of procyanidins on cerebral ischemia-reperfusion injury in rats [J]. China Pharmacist, 2005, 8(7): 541-542.(in Chinese)
[7] 贺 玲. 原花青素对大鼠视网膜缺血再灌注损伤后视神经的保护作用[J]. 新乡医学院学报,2012,29(1):26-28. HE Ling. Protective effect of proanthocyanidin on optic nerversafter experimental retinal ischemia-reperfusion injury in rats [J]. Journal of Xinxiang Medical College, 2012, 29(1): 26-28. (in Chinese)
[8] 权晶晶. 二氯化钴在体外细胞缺氧研究中的应用[J]. 国际口腔医学杂志, 2009,6(4):455-458. QUAN Jing-jing. Application of cobalt chloride in cell hypoxia in vitro research [J]. International Journal of Stomatology, 2009,6(4):455-458.(in Chinese)
[9] WU F J, XUE Y, TANG Q J, et al. Protective effects of cerebrosides from sea cucumber and starfish on the oxidative damage in PC12 cells [J]. J Oleo Sci, 2013, 62(9): 717-727.
[10] AMES A 3RD. Energy requirements of CNS cells as related to their function and to their vulnerability to ischemia: a commentary based on studies on retina [J]. Can J Physiol Pharmacol, 1992, 70 Suppl: S158-S164.
[11] MOZAFFARIEH M, GRIESHABER M C, ORGUL S, et al. The potential value of natural antioxidative treatment in glaucoma [J]. Surv Ophthalmol, 2008, 53(5):479-505.
[12] 原慧萍, 马春阳, 周欣荣, 等. 原花青素对微波诱导视网膜神经节细胞凋亡的拮抗作用[J]. 中国病理生理杂志, 2008,24(4):812-814. YUAN Hui-ping,MA Chun-yang,ZHOU Xin-rong,et al. Antagonism effect of procyanidins on microwave induced cell apoptosis in retinal ganglion cells[J]. Chinese journal of pathophysiology, 2008, 24(4):812-814.(in Chinese)
[13] KANNAN K, JAIN S K. Oxidative stress and apoptosis [J]. Pathophysiology, 2000, 7(3): 153-163.
[14] YANG E, KORSMEYER S J. Molecular thanatopsis: a discourse on the BCL2 family and cell death [J]. Blood, 1996, 88(2):386-401.
[15] DU L, LI Z J, XU J, et al. The anti-tumor activities of cerebrosides derived from sea cucumber Acaudina molpadioides and starfish Asterias amurensis in vitro and in vivo [J]. J Oleo Sci, 2012, 61(6):321-330.
[16] XIE Z, MOIR R D, ROMANO D M, et al. Hypocapnia induces caspase-3 activation and increases Abeta production [J]. Neurodegener Dis, 2004, 1(1):29-37.
[17] MARTINOU J C, YOULE R J. Mitochondria in apoptosis: Bcl-2 family members and mitochondrial dynamics [J]. Dev Cell, 2011, 21(1):92-101.
[18] GROSS A, MCDONNELL J M, KORSMEYER S J. BCL-2 family members and the mitochondria in apoptosis [J]. Genes Dev, 1999, 13(15):1899-1911.
[19] ROY A M, BALIGA M S, ELMETS C A, et al. Grape seed proanthocyanidins induce apoptosis through p53, Bax, and caspase 3 pathways [J]. Neoplasia, 2005, 7(1):24-36.
[20] LIU Z, CHEN J, HUANG W, et al. Ginsenoside Rb1 protects rat retinal ganglion cells against hypoxia and oxidative stress [J]. Mol Med Rep, 2013, 8(5):1397-1403.
[1] 张力三,陈冠峰,陈洁芳,何旭东,胡兴越. 组胺改善癫痫所致记忆形成障碍的实验研究[J]. 浙江大学学报（医学版）, 2017, 46(1): 1-6.
[2] 高思倩,沈咏梅,耿福能,李艳华,高建青. 糖尿病溃疡动物模型的建立及相关治疗研究进展[J]. 浙江大学学报（医学版）, 2017, 46(1): 97-105.
[3] 汪锡婷,唐藕凤,叶夷露,郑鸣之,胡珏,陈忠,钟恺. 藏红花素对小鼠电点燃癫痫的影响[J]. 浙江大学学报（医学版）, 2017, 46(1): 7-14.
[4] 封盛 等. 糖皮质激素受体信号通路在膀胱癌治疗中的作用研究进展[J]. 浙江大学学报（医学版）, 2016, 45(6): 655-660.
[5] 刘军 等. 机体炎症因子和氧化应激标志物介导姜黄素抑制骨性关节炎的作用机制[J]. 浙江大学学报（医学版）, 2016, 45(5): 461-468.
[6] 林先刚 等. 姜黄素对慢性阻塞性肺疾病大鼠肺动脉平滑肌细胞的作用及其机制研究[J]. 浙江大学学报（医学版）, 2016, 45(5): 469-476.
[7] 王结胜 等. 小檗碱抑制肠道二肽基肽酶-Ⅳ的降糖机制研究[J]. 浙江大学学报（医学版）, 2016, 45(5): 486-492.
[8] 肖卫强 等. 不同模式口鼻式吸烟诱导急性肺损伤的实验研究[J]. 浙江大学学报（医学版）, 2016, 45(5): 522-529.
[9] 孟楠楠 等. 甲基丁香酚对变应性鼻炎大鼠鼻黏膜组织中黏蛋白5AC的影响[J]. 浙江大学学报（医学版）, 2016, 45(5): 477-485.
[10] 刘军 等. JAK2/STAT3信号通路介导薯蓣皂苷元对骨性关节炎软骨细胞代谢的影响[J]. 浙江大学学报（医学版）, 2016, 45(5): 453-460.
[11] 白石 等. 雷公藤甲素缓解局灶性脑组织缺血再灌注损伤诱发的炎症及其相关凋亡的研究[J]. 浙江大学学报（医学版）, 2016, 45(5): 493-500.
[12] 曹鹏 等. 双氢青蒿素抗肿瘤分子生物学机制研究进展[J]. 浙江大学学报（医学版）, 2016, 45(5): 501-507.
[13] 何斌 等. 贝伐珠单克隆抗体在难治性子宫颈癌中的应用进展[J]. 浙江大学学报（医学版）, 2016, 45(4): 395-402.
[14] 陶泉玮等. 聚腺苷二磷酸核糖聚合酶抑制剂联合卡铂对人乳腺癌细胞凋亡的影响[J]. 浙江大学学报（医学版）, 2015, 44(5): 506-510.
[15] 陈子博等. 舒尼替尼通过调控转化生长因子β介导的上皮-间质转化抑制卵巢癌细胞转移[J]. 浙江大学学报（医学版）, 2015, 44(5): 479-485.