17β-雌二醇共轭衍生物对大鼠脑缺血再灌注神经损伤的保护作用及机制

发布时间：2018-07-09 09:21

本文选题：17β-雌二醇共轭衍生物 + 核外雌激素受体　；参考：《南华大学》2011年硕士论文

【摘要】：目的：本研究以SD大鼠缺血再灌注神经损伤为模型，探讨17β-雌二醇共轭衍生物(E2-BSA)对脑缺血再灌注神经损伤的保护作用及其机制。方法：通过建立SD大鼠脑缺血再灌注损伤模型，将实验动物分为假手术组(SH)、模型组(IRI)、不同浓度E2-BSA治疗组、10μM E2-BSA+核内雌激素受体抑制剂ICI182780组(ICI)和10μM E2-BSA加PI3K抑制剂LY294002组（LY）。首先通过HE染色观察不同浓度E2-BSA对大鼠缺血再灌注神经损伤的保护作用，筛选出最佳的E2-BSA预保护浓度；然后进一步通过DNA ladder、RT-PCR和western blot观察E2-BSA抗SD大鼠缺血再灌注致神经损伤与核外雌激素受体及PI3K/Akt信号途径的关系，并检测PI3K/Akt下游信号分子Caspase3的活性变化。结果：E2-BSA对大鼠脑缺血再灌注损伤神经细胞起保护作用，，并且较其它浓度，10μM E2-BSA处理后HE染色示大脑皮层神经细胞和海马区神经细胞损伤较轻。westernblot结果显示，脑缺血再灌注损伤可明显抑制神经细胞Akt的活性； E2-BSA可上调脑缺血再灌注损伤后神经细胞Akt的活性， PI3K抑制剂LY294002可抑制E2-BSA对Akt活性的上调作用。DNAladder结果显示，脑缺血再灌注损伤可明显诱导神经细胞凋亡，给以E2-BSA处理后神经细胞凋亡明显减轻，核内雌激素受体抑制剂不能逆转E2-BSA的抗凋亡作用，但可被PI3K抑制剂LY294002所逆转。RT-PCR和westernblot结果显示，脑缺血再灌注损伤可明显促进神经细胞caspase3表达；给以E2-BSA处理后，脑缺血再灌注损伤诱导的caspase3高表达明显减轻，核内雌激素受体抑制剂不能逆转E2-BSA对caspase3高表达的抑制作用，但可被PI3K抑制剂LY294002所逆转。结论：E2-BSA侧脑室注射可有效地起到对脑缺血再灌注损伤大鼠神经细胞的保护作用。E2-BSA的保护机制与核内ER无关，通过核外ER迅速起作用。E2-BSA通过P13K/Akt途径来起到对脑缺血再灌注损伤神经细胞的保护作用。E2-BSA能够迅速通过P13K/Akt途径来抑制下游分子Caspase3的活性来起到对脑缺血再灌注损伤神经细胞的保护作用。
[Abstract]:Aim: to investigate the protective effect and mechanism of 17 尾 -estradiol conjugate derivative (E2-BSA) on cerebral ischemia-reperfusion nerve injury in SD rats. Methods: SD rats were divided into sham-operated group (SH), model group (IRI), 10 渭 M E2-BSA treatment group (ICI182780) and 10 渭 M E2-BSA plus PI3K inhibitor LY294002 (LY). Firstly, the protective effects of different concentrations of E2-BSA on ischemia-reperfusion nerve injury were observed by HE staining, and the best pre-protective concentration of E2-BSA was selected. Then the relationship between E2-BSA and extracellular estrogen receptor and PI3K / Akt signaling pathway was observed by RT-PCR and western blot in SD rats, and the activity of Caspase3 downstream signal molecule of PI3K / Akt was detected. Results compared with other concentrations of 10 渭 M E2-BSA, the brain cortex neurons and hippocampal nerve cells were slightly damaged by HE staining. E2-BSA could up-regulate the activity of Akt in neurons after cerebral ischemia-reperfusion injury, and PI3K inhibitor LY294002 could inhibit the up-regulation of Akt activity by E2-BSA. Cerebral ischemia-reperfusion injury could induce apoptosis of nerve cells, and the apoptosis of neurons was alleviated after treatment with E2-BSA. The inhibitor of estrogen receptor in nucleus could not reverse the anti-apoptosis effect of E2-BSA. However, reverse transcription-polymerase chain reaction (RT-PCR) and westernblot were reversed by LY294002, a PI3K inhibitor. The results of RT-PCR and westernblot showed that cerebral ischemia-reperfusion injury could significantly promote the expression of caspase3 in neurons, and the overexpression of caspase3 induced by cerebral ischemia-reperfusion injury was significantly reduced after treatment with E2-BSA. E2-BSA could not reverse the high expression of caspase3, but could be reversed by PI3K inhibitor LY294002. Conclusion the ICV injection of% E2-BSA can effectively protect neurons from cerebral ischemia-reperfusion injury. The protective mechanism of E2-BSA is not related to ER in nucleus. E2-BSA can protect neurons from cerebral ischemia-reperfusion injury through P13K / Akt pathway through E2-BSA. E2-BSA can inhibit the activity of Caspase3 by P13K / Akt pathway to induce cerebral ischemia-reperfusion injury. The protective effect of nerve cells.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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