解剖学报

›› 2016, Vol. 47 ›› Issue (1): 28-33.

• 神经生物学 • 上一篇    下一篇

龙胆苦苷对新生大鼠氧糖剥夺再灌注损伤海马神经元Caspase-3、Bax和Bcl-2表达的影响

王建中1,雷有杰2,蒋红英3,王福青1   

  1. 1. 漯河医学高等专科学校
    2. 周口职业技术学院医学院
    3. 许昌学院公共体育教学部
  • 收稿日期:2015-07-02 修回日期:2015-09-01 出版日期:2016-02-06 发布日期:2016-02-06
  • 通讯作者: 王建中 E-mail:wangjzjp@163.com

Effects of gentiopicroside on Caspase-3, Bax and Bcl-2 in neonatal rat hippocampal nurons following oxygen-glucose deprivation and reperfusion injury

  • Received:2015-07-02 Revised:2015-09-01 Online:2016-02-06 Published:2016-02-06

摘要： 目的 研究龙胆苦苷（Gentiopicroside，GP）对新生大鼠氧糖剥夺再灌注损伤海马神经元的抗凋亡保护作用及机制。方法 采用无糖培养基加缺氧法建立原代新生大鼠海马神经元氧糖剥夺再灌注损伤模型，同时给予不同浓度的龙胆苦苷进行干预。应用Hoechst 33342染色法检测神经细胞的凋亡，并计算凋亡率；化学比色法测定神经细胞乳酸脱氢酶（LDH）漏出率；RT-PCR和Western-blot技术检测凋亡相关因子Caspase-3、 Bax 和 Bcl-2 mRNA和蛋白的表达。结果 与正常组比较，模型组Hoechst 33342染色神经元凋亡率明显升高，LDH漏出率增加，Caspase-3、 Bax mRNA和蛋白的表达水平上调，Bcl-2 mRNA和蛋白的表达水平下降。与模型组比较，龙胆苦苷（40、20、10mg/L）可显著降低氧糖剥夺再灌注损伤神经元的凋亡率和LDH漏出率；龙胆苦苷（40mg/L）可明显抑制Caspase-3、 Bax mRNA和蛋白的表达，升高Bcl-2 mRNA和蛋白的表达。结论 龙胆苦苷对新生大鼠海马神经元氧糖剥夺再灌注损伤所诱发的神经细胞凋亡具有保护作用，其机制可能与上调Bcl-2表达，抑制Caspase-3、Bax表达有关。

关键词: 龙胆苦苷, 氧糖剥夺再灌注损伤, Caspase-3, Bax, Bcl-2

Abstract: Objective To study the anti-apoptosis effects of gentiopicroside in neonatal rat hippocampal neurons following oxygen-glucose deprivation and reperfusion injury and elucidated the related mechanisms. Methods The neonatal rat hippocampal neurons were pretreated with gentiopicroside 24h before exposed to oxygen-glucose deprivation and reperfusion injury were used as an in vitro model of ischemia and reperfusion. The neuron morphological change and apoptosis rate were evaluated by Hoechst 33342 staining. The nerve cell lactate dehydrogenase (LDH) leakage rate was detected by spectrophotometry. The expression levels of Caspase-3, Bax and Bcl-2 mRNA and protein were detected by RT-PCR and Western-blot assay respectively. Results Compared with the control group, oxygen-glucose deprivation and reperfusion injury model group could significantly enhance the apoptosis rate of nerve cells, increase the LDH leakage rate. Compared with the model group, pretreated with gentiopicroside (40, 20, 10mg/L) could significantly attenuated neuronal damage, with evidence of decreased neurons apoptosis rate and LDH leakage rate. Compare with the model group, pretreated with gentiopicroside (40 mg/L) could effectively down regulate the expression of Caspase-3 and Bax in mRNA and protein level, and up regulate the expression of Bcl-2 level. Conclusion Gentiopicroside preconditioning could prevent neurons from oxygen-glucose deprivation and reperfusion injury, and the mechanism may be mediated by up regulated Bcl-2 and down regulated Caspase-3 and Bax.

Key words: Gentiopicroside, oxygen-glucose deprivation and reperfusion injury, Caspase-3, Bax, Bcl-2