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  

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

作　　者：王建中[1] 雷有杰 蒋红英[3] 王福青[1] 

机构地区：[1]漯河医学高等专科学校解剖学教研室,河南漯河462002 [2]周口职业技术学院医学院人体教研室,河南周口466000 [3]许昌学院公共体育教学部,河南许昌461000

出　　处：《解剖学报》2016年第1期28-33,共6页Acta Anatomica Sinica

摘　　要：目的探讨龙胆苦苷(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 blotting技术检测凋亡相关因子Caspase-3、Bax和Bcl-2 m RNA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 m RNA和蛋白的表达水平上调,Bcl-2 m RNA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 m RNA和蛋白的表达,升高Bcl-2 m RNA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。Objective To study the anti-apoptosis effects of gentiopicroside in neonatal rat hippocampal neurons following oxygen-glucose deprivation and reperfusion injury. Methods The neonatal rat hippocampal neurons were pretreated with gentiopicroside 24hours before exposed to oxygen-glucose deprivation. 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 blotting assay. Results Compared with the control group, oxygen-glucose deprivation and reperfusion injury model group significantly enhanced the apoptosis rate of nerve cells, and increased the LDH leakage rate. Compared with the model group, the group pretreated with gentiopicroside （40, 20, 10mg/L） significantly attenuated neuronal damage, with evidence of decreased neurons apoptosis rate and LDH leakage rate. Compared with the model group, the group pretreated with gentiopicroside （40mg/L） effectively down regulated the expression of Caspase-3 and Bax in mRNA and protein level, and up regulated the expression of Bcl-2 level. Conclusion Gentiopicroside preconditioning can prevent neurons from oxygen-glucose deprivation and reperfusion injury, which may be mediated by up regulating Bcl-2 and down regulating Caspase-3 and Bax.

关 键 词：龙胆苦苷 氧糖剥夺再灌注损伤 反转录-聚合酶链反应 大鼠 

分 类 号：R743[医药卫生—神经病学与精神病学] R966[医药卫生—临床医学]