目的:本研究在成功建立AD体外模型基础上,探讨补肾化痰益智法治疗AD的作用机制。方法:运用Aβ1-42作用于PC-12细胞,制作AD神经元细胞模型,并用补肾化痰益智方含药血清进行干预,以吲哚美辛为对照。镜下观察细胞形态,MTT法检测PC-12细胞存活率,观察补肾化痰益智方对神经元细胞保护作用。RT-PCR法检测APP mRNA的表达,观察补肾化痰益智方抗Aβ神经毒性作用。结果:2.5 mmol·L-1聚集态Aβ1-42使细胞活力明显下降;RT-PCR法对APP mRNA表达进行半定量分析发现,经Aβ1-42诱导各组APP mRNA表达上调,补肾化痰益智法可明显降低其表达(P〈0.01)。结论:补肾化痰益智法对神经细胞具有较好的保护作用,能够抑制APP mRNA的表达,降低毒性作用。
This study successfully established in v itro model of Alzheimerˊs disease ( AD ) in order to investigate effects and mechanisms on AD by Bushen-Huatan-Yizhi Recipe (BSHTYZR, replenishing kidney-essence and removing phlegm to benefit wisdom ) . The PC-12 cells affected by Aβ1-42 incubated were produced for neuron cell model of AD . Then , they were intervened by the serum containing BSHTYZR , which were compared to the serum containing indomethacin . The cell morphology was observed . The neuronal survival was assessed by counting MTT assay . The expression of APP mRNA was detected with RT-PCR . And the protective effect of BSHTYZR on neuronal cells and neurotoxic effects of anti-Aβ was observed . The results showed that 2 . 5 mmol/L Aβ1-42 can significantly decrease the cell viability . The RT-PCR was used on semi-quantitative analysis for expression of APPmRNA. It showed that expression of APP mRNA was increased in each group, and the BSHTYZR group can reduce the expression significantly ( P 〈 0 . 01 ) . It was concluded that the BSHTYZR method can protect nerve cells , suppress the expression of APP mRNA , and reduce the toxic effects .