复智散对神经细胞胞体面积和轴突长度变化的影响(英文)

Effect of fuzhisan on the area of neurosome and the length of axon

背景:自制中药复智散经已往的实验证实可延缓大鼠的自然老化过程提示该药可能具有对抗衰老作用。目的:观察复智散培养神经细胞最佳有效浓度对神经母细胞瘤株SHSY5Y细胞形态学改变的影响。设计:重复测量设计。材料:实验于2002-06/2003-04在首都医科大学宣武医院北京脑老化重点实验室完成。自制中药复智散(菖蒲、远至等6味中药组成)由哈尔滨医科大学第一临床医学院神经科王德生教授和上海东方医院神经科徐晓云教授共同组方,淀粉样β蛋白25-35片段,SH-SY5Y神经母细胞瘤株。方法:用不同浓度的复智散孵育神经母细胞瘤株SH-SY5Y细胞,利用噻唑蓝比色法测定细胞存活率,制定量-效关系曲线,寻找最佳药物浓度;6孔板培养细胞分为正常对照组、淀粉样β蛋白25-3525μmol/L组、淀粉样β蛋白25-3525μmol/L+复智散45×10-3g/L组和复智散45×10-3g/L组,孵育24h,观察细胞形态学改变,测定神经细胞的胞体面积和轴突长度。主要观察指标:①各组SH-SY5Y细胞噻唑蓝代谢率的变化。②各组神经细胞胞体面积及轴突长度。结果:①复智散可增进SH-SY5Y细胞的存活,最佳有效浓度为45×10-3g/L。②SH-SY5Y细胞胞体面积和轴突长度:淀粉样β蛋白25-3525μmol/L组明显低于正常对照组犤(505.5±122.36),(599.8±141.25)μm2;(26.0±13.97),(36.5±15.58)μm,(t=3.903,3.447,P=0.000)犦。复智散45×10-3g/L组与淀粉样β蛋白25-3525μmol/L+复智散45×10-3g/L组均明显高于淀粉样β蛋白25-3525μmol/L组犤(918.3±178.34),(896.6±257.14),(505.5±122.36)μm2;(96.8±43.31),(88.3±30.23),(26.0±13.97)μm,(t=10.922,14.172,P=0.000)犦。结论:在淀粉样β蛋白25-35损伤条件下复智散依旧有促进培养神经细胞存活的作用,表现在增进细胞胞体面积的增长和轴突的延伸方面。

BACKGROUND： It has been verified in the experiments over the past that the self-prepared Chinese herb, fu, zhisan can retard natural aging in rats, suggesting that such drug acts on anti-aging. OBJECTIVE： To observe the effect of optimum effective concentration of nerve cell cultured with fuzhisan on morphological alternation of neuroblastoma SH-SY5Y cell. DESIGN： Repeated measurement. MATERIALS： The experiment was performed in Beijing Key Laboratory Room of Cerebral Aging of Xuanwu Hospital Affiliated to Capital University of Medical Sciences from June 2002 to April 2003. Self-prepared Chinese herb, fu.zhisan [composed of 6 herbs, such as shichangbu （Rhizoma Acori Graminei）, yuan.zhi （Radix Polygalae）, etc.I was co-developed by Prof. Wang De-shen from Department of Neurology of First Clinical Medical College of Harbin Medical University and Prof. Xu Xiao-yun from Department of Neurology of Shanghai Odental Hospital. In addition, amyloid β- protein 25-35 segment and SH-SY5Y neuroblastoma were provided. METHODS： Fuzhisan of various concentrations were used for incubation of neuroblastoma SH-SY5Y cell. Thiazolyl blue （MTT）, colorimetric method was used to determine the cell survival rate. Dose-effect relationship curve was drawn up to search optimum drug concentration. The cells cultured with 6-pore plate were divided into normal control, amyloid 13-protein 25 -35 25μmol/L group, amyloid β-protein 25-35 25 μmol/L ± fuzhisan 45×10^-3 g,/L groups andfuzhisan 45×10^-3 g,/L group. They were incubated for 24 hours to observe cell morphological alternation and determine neurnsome area and axon length. MAIN OUTCOME MEASURES： ① Changes in MTT metabolic rate of SH-SY5Y cell in every group. ② Neurosome area and axonal length of every group. RESULTS： ① Fuzhisaa enhanced the survival of SH-SY5Y cell at the optimum concentration of 45×10^-3g/L. ② Area of neurosome and axonal length of every group： Those in amyloid β-protein 25-35 25 μmol/L group were decreased remarkably than the normal control [（505,5 ±122.36）, （599.8±141.25） μm2; （26.0±13.97）, （36.5±15.58）μm, （t=3.903, 3.447, P=0.000）]. Those infuzhiscm 45×10^-3g/L group and amyloid β-pro- tein 25-35 25 μmol/L ＋ fuzhisan 45×10^-3 g/L group were increased remarkably than amyloid β-protein 25-35 25 μxmol/L group [（918.3±178.34）, （896.6±257.14）, （505.5±122.36） μxm^2; （96.8±43.31）, （88.3±30.23）, （26.0±13.97） μxm, （t=10.922, 14.172, P=0.000）]. CONCLUSION： With injury, of amyloid β-protein 25-35, fuzhisan still enhances the survival of cuhured nerve cell, manifested as promoting the increase of neurosome area and axonal extension,