清脑益智方含药脑脊液对缺氧复氧皮层神经元细胞重塑相关指标的影响

目的:探究清脑益智方通过促神经元突触重塑治疗血管性痴呆的机制.方法:原代培养皮层神经元,将神经元细胞按照1×106/mL的密度接种于96孔细胞培养板,将细胞分为正常组、缺氧复氧组、正常脑脊液组(正常CSF 10％的元血清培养液)、清脑益智方15.7 g.kg-1给药,连续3d制备的含药脑脊液高剂量组(清脑益智方-CSF 10％的元血清培养液)、清脑益智方含药脑脊液低剂量组(清脑益智方-CSF 5％的元血清培养液)、恩必普组含75 mg'kg-1 ig3 d制备的脑脊液5％的元血清培养液.除正常细胞组外,其余各组均在添加相应干预措施后进行缺氧24 h复氧24 h处理.采用ELISA法检测细胞上清液中突触素(synapsin,SYN)、生长相关蛋白-43(growth associated protein-43,GAP-43)、微管相关蛋白2(microtubule associated protein 2,MAP-2)等突触重塑标志物的表达.结果:清脑益智方含药脑脊液能够促进缺氧复氧皮层神经元突触重塑相关指标SYN,GAP-43,MAP-2的表达,并且清脑益智方含药脑脊液高剂量组与低剂量组的上述3个指标表达水平均明显高于缺氧复氧组,且具有显著的统计学差异(P＜0.01).结论:清脑益智方可通过SYN,MAP-2,GAP-43等突触重塑标志物的表达而促进神经元重塑.

Effect of Qingnao Yizhi Decoction on Synaptic Remodeling Related Indexes

Objective: To investigate the possible mechanism of Qingnao Yizhi(QNYZ)decoction on vascular dementia(VD) through testing the effect of vcerebrospinal fluid which contained Qingnao Yizhi decoction on synaptic remodeling so as to provide more scientific evidence for the clinical medication on VD patients. Method: The primary cultured rat cortical neurons were vaccinated in 96-well cell culture plates and the vaccination concentration was 1×10⁶/mL.And the cells were divided into control group,H/R group,cerebrospinal fluid(CSF) group,high dose CSF containing QNYZ group(QNYZ-H),low dose CSF containing QNYZ group(QNYZ-L) and CSF containing butylphthalide(NBP) group(except the control group,the other groups were firstly dealt with hypoxia for 24 hours and then received reoxygenation for 24 hours). The concentration of synaptic remodeling markers such as synapsin,microtubule associated protein 2 and growth associated protein-43 of the supernatant were assayed by the ELISA way. Result: The concentration of synaptic remodeling markers such as synapsin(SYN),microtuble associated protein 2(MAP-2) and growth associated protein-43(GAP-43) of the supernatant of the QN-H,QN-L groups increased,and the three indices'level are obviously higher than these in the H/R group(P<0.01). Conclusion: Qingnao Yizhi decoction could improve the synaptic remodeling of the H/R CNC through regulating the expression of SYN,MAP-2 and GAP-43.