食品科学 ›› 2018, Vol. 39 ›› Issue (3): 222-228.doi: 10.7506/spkx1002-6630-201803034

• 营养卫生 • 上一篇    下一篇

白扁豆多糖对神经细胞缺氧性凋亡的保护机制

张贤益1,李文娟1,钟 亮1,潘 猛1,汤小芳1,姚于飞2,*   

  1. 1.南昌大学 食品科学与技术国家重点实验室,江西 南昌 330047;2.中国人民解放军第九四医院,江西 南昌 330000
  • 出版日期:2018-02-15 发布日期:2018-01-30
  • 基金资助:
    南京军区面上项目(15MS086);江西省自然科学基金重点项目(20171ACB21016)

Protective Mechanism of Polysaccharides from Dolichos Bean Seeds (Dolichos lablab L.) on Hypoxia-Induced Neuronal Apoptosis

ZHANG Xianyi1, LI Wenjuan1, ZHONG Liang1, PAN Meng1,TANG Xiaofang1, YAO Yufei2,*   

  1. 1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China; 2. Chinese People’s Liberation Army No. 94 Hospital, Nanchang 330000, China
  • Online:2018-02-15 Published:2018-01-30

摘要: 目的:模拟体内环境,研究白扁豆多糖(dolichos bean seed polysaccharide,DBSP)对胎鼠大脑 皮层神经细胞缺氧性凋亡的保护机制。方法:原代培养胎鼠大脑皮层神经细胞;建立缺氧/复氧(anoxia/ reoxygenation,A/R)损伤模型;实验分4 组:对照组、A/R组、白扁豆多糖预处理组(DBSP+A/R组)、阻断剂组 (LY294002+DBSP+A/R组);用比色法检测细胞活性与乳酸脱氢酶的释放水平;流式细胞仪分析测定Ca2+ 相对浓度、活性氧、线粒体膜电位和细胞凋亡的变化;Western blot法检测总蛋白激酶B(total protein kinase B, T-Akt)和磷酸化蛋白激酶B(phosphorylated protein kinase B,p-Akt)蛋白表达。结果:从细胞活性来看,A/R组比 对照组明显降低(P<0.01),表明A/R对神经细胞产生了损伤。同时,与A/R组相比,DBSP+A/R组细胞活性显著 上升、细胞凋亡明显减少(P<0.01),乳酸脱氢酶的释放水平下降,表明DBSP对受A/R损伤的神经细胞产生保护 作用,进一步结果显示,与A/R组相比,DBSP+A/R组中Ca2+相对浓度和活性氧生成显著降低、显著增加线粒体膜 电位和p-Akt蛋白表达(P<0.01)。而与DBSP+A/R组相比,LY294002+DBSP+A/R组中p-Akt蛋白表达显著减少 (P<0.01),同时细胞活性与线粒体膜电位下降,且LDH的释放水平、Ca2+相对浓度、活性氧、细胞凋亡明显增加 (P<0.05),表明DBSP对神经细胞的保护作用受到PI3K-Akt信号通路抑制剂LY294002的抑制作用而被削弱。结 论:DBSP可通过PI3K-Akt信号转导通路抑制神经细胞的缺氧性凋亡。

关键词: 白扁豆多糖, 神经细胞, 缺氧/复氧, 凋亡作用, PI3K-Akt信号通路

Abstract: Objective: The aim of this study was to explore the protective effect of dolichos bean seed polysaccharides (DBSP) against anoxia/reoxygenation (A/R) injury and its potential mechanism of action in neurons. Methods: Primary cultured cerebral cortical neurons were randomly divided into four groups including control group, A/R group, DBSP + A/R group and DBSP + LY294002 + A/R group. Cell survival rate and lactate dehydrogenase (LDH) release were determined. Apoptosis, mitochondrial membrane potential and Ca2+ concentration were determined by flow cytometry analysis. The expression of T-Akt and p-Akt protein was evaluated by Western blot. Results: Compared with the control group, cell survival rate of the A/R group was decreased (P < 0.01) and LDH release, Ca2+ concentration and apoptotic rate were significantly increased (P < 0.01). Compared with the A/R group, cell survival rate of the DBSP + A/R group was increased, and LDH release, Ca2+ concentration and apoptotic rate were significantly decreased (P < 0.01), suggesting that DBSP could protect neurons against A/R injury. Mitochondrial membrane potential and the expression of p-Akt protein increased significantly in the DBSP + A/R group when compared with the A/R group (P < 0.01). The expression of p-Akt protein after treatment with LY294002 (a potent inhibitor of the PI3K-Akt signaling pathway) + DBSP + A/R was attenuated in neurons when compared with A/R + DBSP treatment (P < 0.05). Further experimental results showed that the treatment of cells with LY294002 completely blocked the expression of p-Akt protein, and significantly attenuated the protective effect of DBSP on neurons subjected to A/R. Conclusion: The protective mechanism of polysaccharides from dolichos bean seeds on neuronal cells is closely related to the PI3K-Akt signaling pathway.

Key words: dolichos bean seed polysaccharide, neuronal cells, anoxia/reoxygenation, apoptotic effect, PI3K-Akt signaling pathway

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