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目的探讨表没食子儿茶素没食子酸酯(EGCG)对丙烯酰胺(ACR)致小脑颗粒神经元(CGNs)损伤的保护作用。方法采用体外细胞培养的方法,建立ACR损伤CGNs的体外模型。经不同浓度EGCG(5、10、25、50、100 mol/L)预处理24h后,加入ACR(5 mmol/L)染毒,24 h后用四甲基偶氮唑盐(MTT)比色法检测细胞存活率,倒置显微镜下观察细胞形态学变化,测定细胞内超氧化物歧化酶(SOD)活性及丙二醛(MDA)的含量,Hochest 33342染色观察凋亡时细胞核形态的变化,末端脱氧核苷酸转移酶介导的原位缺口末端标记(TUNEL)法检测细胞凋亡指数。结果同ACR损伤组比较,终浓度为10、25、50 mol/L的EGCG能减轻ACR引起的CGNs损伤,可明显提高细胞的存活率(P<0.05),SOD活性增高,MDA含量降低(P<0.05),Hoechst33342染色和TUNEL荧光标记显示,细胞核固缩、致密浓染现象较ACR损伤模型组改善明显,凋亡指数下降(P<0.05),且呈现剂量依赖趋势,而终浓度为5、100 mol/L EGCG组较正常组无明显变化。结论 EGCG在一定剂量范围内对ACR氧化损伤的CGNs有防护作用。[营养学报,2012,34(4):362-367]
Objective To investigate the protective effect of epigallocatechin gallate (EGCG) on damage of cerebellar granule neurons (CGNs) induced by acrylamide (ACR) in rats. Methods The in vitro cell culture method was used to establish an in vitro model of ACN-injured CGNs. After being pretreated with different concentrations of EGCG (5, 10, 25, 50, 100 mol / L) for 24 h, ACR (5 mmol / L) was added and the cells were stained with MTT Cell viability was measured by inverted microscope. Morphological changes of cells were observed under inverted microscope. The activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA) were measured. Hochest 33342 staining was used to observe the morphological changes of nuclei during apoptosis. Deoxyribonucleotide transferase mediated in situ nick end labeling (TUNEL) method was used to detect apoptotic index. Results Compared with ACR injury group, EGCG at the final concentration of 10, 25 and 50 mol / L could reduce the damage of CGNs induced by ACR, and significantly improve the cell survival rate (P <0.05), SOD activity and MDA content <0.05). Hoechst33342 staining and TUNEL fluorescent labeling showed that the phenomenon of nuclear condensation and dense staining significantly improved compared with ACR injury model (P <0.05), and the apoptotic index decreased in a dose-dependent manner. The final concentration was 5, 100 mol / L EGCG group than the normal group no significant change. Conclusion EGCG has a protective effect on ACN oxidative damage of CGNs in a certain dose range. [Journal of Nutrition, 2012,34 (4): 362-367]