盐酸多奈哌齐对D-半乳糖致原代培养神经元损伤的保护作用及其机制

Mechanism of Donepezil Hydrochloride on Cellular Injury of Primary Cultured Neurons Induced by D-galactose

目的:探讨盐酸多奈哌齐(安理申)防治D-半乳糖致原代培养神经元损伤的作用机制。方法:在原代培养第6d,以50mM D-半乳糖(D-gal)作用神经元72h造成自由基损伤模型,用10％盐酸多奈哌齐药物血清防治损伤。倒置显微镜下观察神经元的生长发育形态,MTT法在酶标仪上检测神经元代谢率,PI染色在流式细胞仪上检测神经元凋亡构成比,HE染色观察神经元形态、形态计量神经突起密度,RT-PCR检测糖醛还原酶信使RNA(AR-mRNA)的含量。t检验,比较正常对照神经元、受D-gal攻击神经元和受D-gal攻击同时得到盐酸多奈哌齐防治神经元之间的差别。结果与讨论:受D-gal攻击而受损伤的神经元,生长发育显著迟缓,盐酸多奈哌齐防治使其明显恢复;代谢率从0.762±0.030(n=33)降低到0.543±0.064(n=11,P<0.01),盐酸多奈哌齐防治使其回升到0.652±0.028(n=11,P<0.01);凋亡构成比从0.060±0.029(n=19)增高到0.356±0.215(n=19,P<0.01),盐酸多奈哌齐防治使其回降到0.154±0.130(n=19,P<0.01);出现变性坏死的形态变化,盐酸多奈哌齐防治使其好转,但神经突起浆质中出现均匀、细小的颗粒;神经突起密度从0.557±0 0422(n=10)降低到0.468±0.0330(n=10,P<0.01),盐酸多奈哌齐防治使其回升到0.481±0.0387(η=10,P>0.05);无AR-mRNA的表达。

Aim: To research the mechanism of donepezil hydrochloride for treating neural in primary neurons induced by D-galac-tose. Method: At day 6 in primary rat neuron culture. 50 mM D-galactose and 10% donepezil hydrochloride-contained serum were added into the medium for the last 72 h. The neural growth and development were observed ender an inverted microscope, the neural metabolism rate was calculated by MTT and immuno-enzyme assay, the neural apoptosis ratio was counted by PI stain and flow mi-crofluorimtry, the neurite density was measured by HE stain and microphotometry, the content of aldose reductase-mRNA was detested by RT-PCR. The difference between the control neurons without D-galactose by t-test, the injured neurons induced by 50 mM D-galactose growth and development in injurd neurons were rescued with a donepezil hydrochloride, the neural. Result and Discussion: The retard neural growth and development in injured neurons were rescued with donepezil hydrochloride; the neural metabolism rate of injured meurons was declined from 0. 762?