白藜芦醇对β-淀粉样蛋白(Aβ_(25～35))诱导的原代培养大鼠皮层神经元损伤的作用研究

The effects of resveratrol on primer cultured cortex neurons injury induced by β-amyloid protein

目的研究白藜芦醇(Res)对β-淀粉样蛋白(Aβ_(25～35))诱导的原代培养大鼠皮层神经元损伤的保护作用及可能的作用机制。方法用原代培养SD大鼠皮层神经元,分为实验组(a,b,c,d),治疗组(e,f,g,h)和空白对照组,实验组分别暴露浓度为5,10,20,40μmol/LAβ_(25～35);治疗组为神经元首先被给于5,10,20,40μmol/LRes作用24h,后再给于20μmol/LAβ_(25～35),孵育24h,倒置显微镜下观察神经元形态学的变化,用四唑盐(MTT)比色试验检测神经元存活和生长情况,应用比色法测定吸光值检测细胞培养上清液中丙二醛(MDA)、谷胱甘肽(GSH)的含量。结果Aβ_(25～35)在终浓度为20μmol/L时神经元形态有明显的变化,细胞立体感减弱,胞体内空泡形成,细胞突起内可见黑色颗粒状物;细胞的存活能力明显下降,与对照组相比较有显著性差异(P<0.05);Res在终浓度40μmol/L时,能明显提高细胞的存活能力。Aβ_(25～35)可以明显升高细胞培养上清液中MDA含量,降低GSH的含量;Res可使细胞培养上清液中MDA含量下降,GSH含量增加。结论Aβ_(25～35)诱导了原代培养皮层神经元变性、死亡,增加了神经元的脂质过氧化,Res可通过抑制脂质过氧化对皮层神经元起到保护作用。

Objective ： To study the effects and the probable mechanisms of resveratrol on primary cultured cortex neurons injury induced by β-amyloid protein. Methods： Morphological study of neuron cell by inverted microscopy, the rate of cell viability by MTT test; Themalondialdehydeacid （MDA）, glutathione （GSH） were measured by chromatometry. Results： After incubated with Aβ25-35 20/μmol/L for 24h, the morphology of neurons was significantly varied. Under inverted microscope. stereognosis of neurons died down, there were blank bulb in the neurons, dark particle were seen in the neuritis; the result of MTT test showed that the survival rate of neurons was obviously decreased. Res 40μmol/L increased the survival rate of neurons. Aβ25～35 dose-dependently increased the level of MDA, decreased the level of GSH. Res dose-dependently increased the level of GSH, decreased the level of MDA. Conclusion： Aβ can induce the death and degeneration of primer cultured cortex neurons, increase the level of lipid peroxidative, resveratrol can protect the neuron by reducing the level lipid peroxidative.