摘要
目的考察Aβ25-35对原代培养大鼠皮质神经元的损伤作用。方法将原代培养的大鼠皮质神经元分为对照组、谷氨酸(glutamate,Glu)组及3个浓度的Aβ25-35组,每组各6孔。观察神经元的形态改变,并测定细胞存活率及培养液中乳酸脱氢酶(LDH)的活力值。结果与对照组比较,Glu组可造成大鼠皮质神经元明显损伤,形态发生显著改变,细胞存活率为(53.1±5.5)%(P<0.01),并且培养液中LDH漏出显著增多,LDH的活力值为(39.9±2.9)U/gProt(P<0.01);Aβ25-35三个剂量组神经元存活率均显著降低,分别为(69.3±5.9)%(P<0.01)、(52.7±3.0)%(P<0.01)和(33.2±1.8)%(P<0.01),Aβ25-35低、中剂量组培养液中LDH漏出未见明显改变,LDH活力值分别为(23.5±1.4)U/gProt和(24.7±1.3)U/gProt(P>0.05),Aβ25-35高剂量组培养液中LDH漏出显著升高,LDH活力值为(38.8±2.0)U/gProt(P<0.01),与Glu组相当(P>0.05)。结论 Aβ25-35能剂量依赖性地损伤皮质神经元,1μmol/LAβ25-35剂量组处理24h后细胞存活率可降低50%左右,可用于Aβ25-35体外诱导阿尔茨海默病细胞模型的建立。
Objective To observe the damage effects of Aβ25-35 on primary rat cortical neurons. Methods Rat cortical neurons were cultured and divided into 5 groups： the normal group （control） , the Glu group, and 3 Aβ25-35 groups at ascending dosage, in each of which there were 6 wells of neurons. Morphological alterations of neurons were observed under inverted microscope after treatment, the neuron viability was detected, while LDH release was measured using LDH activity Kit. Results In contrast to controls, neurons in Glu group suffered significant damage with obvious morphological alterations, neuron viability rate was （53. 1 ± 5. 5 ）% , LDH release increased and LDH viability rate was （39. 9 ± 2.9）U/g Prot with difference of statistical significance （P 〈 0. 01 ）. Neuron viability was found to have decreased in all Aβ25-35 groups, they were （69.3±5.9）% （P〈0.01）, （52.7 ±3.0）% （P〈0.01）, and （33.2±1.8）% （P〈0.01） respectively. There were no significant changes of LDH release found in both low and medium Aβ25-35 groups, except in the high Aβ25-35 group. LDH viability rates were （23.5±1.4）U/g Prot and （24.7 ±1.3 ）U/g Prot （P 〉 0. 05 ） respectively. LDH activity in culture medium was （38.8 ±2.0） U/g Prot, which was higher than that in controls （P 〈0. 01 ）, but equivalent to that in Glu group （P 〉 0.05）. Conclusion Cultured rat cortical neurons can be impaired by Aβ25-35 in the dose relevant manner. Neuron viability decreasmay by about 50% after being treated with 1μmol/L Aβ25-35 for 24 h, which can be applied as an Alzheimerg disease cell model in vitro.
出处
《华北国防医药》
2010年第4期301-304,共4页
Medical Journal of Beijing Military Region
