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Journal of ZheJiang University(Medical Science)  2015, Vol. 44 Issue (1): 24-29    DOI: 10.3785/j.issn.1008-9292.2015.01.004
    
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
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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 wordsVITIS VINIFERA/chemistry      Proanthocyanidins/pharmacology      Cobalt/pharmacology      Retinal ganglion cells/drug effects      Anoxia/therapy     
Received: 20 August 2014     
CLC:  R96  
Cite this article:

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.

URL:

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


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

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 

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