CAJ | 학술논문

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SH-SY5Y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25-35 25μmol/L组、淀粉样β蛋白25-35 25μmol/L+复智散45×10-3g/L组和复智散45×10-3g/L组,孵育24 h,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45×10-1g/L.②SH-SY5Y细胞胞体面积和轴突长度:淀粉样β蛋白25-35 25 μmol/L组明显低于正常对照组[(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)].复智散45×10-3 g/L组与淀粉样β蛋白25-35 25 μmol/L+复智散45×1003g/L组均明显高于淀粉样β蛋白25-35 25μmol/L组[(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)].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.
배경:자제중약복지산경이왕적실험증실가연완대서적자연노화과정,제시해약가능구유대항쇠로작용.목적:관찰복지산배양신경세포최가유효농도대신경모세포류주SH-SY5Y세포형태학개변적영향.설계:중복측량설계.재료:실험우2002-06/2003-04재수도의과대학선무의원북경뇌노화중점실험실완성.자제중약복지산(창포、원지등6미중약조성)유합이빈의과대학제일림상의학원신경과왕덕생교수화상해동방의원신경과서효운교수공동조방,정분양β단백25-35편단,SH-SY5Y신경모세포류주.방법:용불동농도적복지산부육신경모세포류주SH-SY5Y세포,이용새서람비색법측정세포존활솔,제정량-효관계곡선,심조최가약물농도;6공판배양세포분위정상대조조、정분양β단백25-35 25μmol/L조、정분양β단백25-35 25μmol/L+복지산45×10-3g/L조화복지산45×10-3g/L조,부육24 h,관찰세포형태학개변,측정신경세포적포체면적화축돌장도.주요관찰지표:①각조SH-SY5Y세포새서람대사솔적변화.②각조신경세포포체면적급축돌장도.결과:①복지산가증진SH-SY5Y세포적존활,최가유효농도위45×10-1g/L.②SH-SY5Y세포포체면적화축돌장도:정분양β단백25-35 25 μmol/L조명현저우정상대조조[(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)].복지산45×10-3 g/L조여정분양β단백25-35 25 μmol/L+복지산45×1003g/L조균명현고우정분양β단백25-35 25μmol/L조[(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)].결론:재정분양β단백25-35손상조건하복지산의구유촉진배양신경세포존활적작용,표현재증진세포포체면적적증장화축돌적연신방면.
BACKGROUND: It has been verified in the experiments over the past that the self-prepared Chinese herb, fuzhisan 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, fuzhisan [composed of 6 herbs, such as shichangbu (Rhizoma Acori Graminei), yuanzhi (Radix Polygalae), etc.] 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 Oriental Hospital. In addition, amyloid βprotein 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 β-protein 25-35 25 μmol/L group, amyloid β-protein 25-35 25 μmol/L + fuzhisan 45×10-3 g/L groups and fuzhisan 45×10-3 g/L group. They were incubated for 24 hours to observe cell morphological alternation and determine neurosome area and axon length.ery group.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 in fuzhisan 45×10-3g/L group and amyloid β-protein 25-35 25 μmol/L + fuzhisan 45×10-3 g/L group were increased remarkably than amyloid β-protein 25-35 25 μmol/L group [(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)].CONCLUSION: With injury of amyloid β-protein 25-35, fuzhisan still enhances the survival of cultured nerve cell, manifested as promoting the increase of neurosome area and axonal extension.