CAJ | 학술논문

目的：探讨茶氨酸对神经干细胞增殖和分化的影响。方法采用机械吹打结合200目钢网滤过的方法，从12．5 d 的昆明胎鼠大脑中分离 NSCs 并培养；用巢蛋白（nestin）免疫荧光染色鉴定 NSC；加入不同浓度茶氨酸（theanine），MTS 法检测不同时间点活性，选定茶氨酸最终加药浓度（100μmol／L）；将 NSCs 分为两组：对照组、加药组（茶氨酸浓度为100μmol／L）；培养后进行 BrdU 染色和微管相关蛋白2（MAP2）、胶质纤维酸性蛋白（GFAP）免疫荧光染色；利用 Q-PCR 以及 Westen blotting 检测 MAP2和 GFAP 表达。结果100μmol／L 茶氨酸处理组MTS 检测的吸光度值显著增高（P ＜0．05）；加药组 BrdU 阳性细胞比率、MAP2阳性细胞比率明显高于对照组（P ＜0．05），两组 GFAP 阳性细胞比率无统计学差异；加药组 MAP2表达明显高于对照组（P ＜0．05），两组 GFAP表达无统计学差异。结论茶氨酸可促进 NSCs 增殖及其向神经元方向分化。
목적：탐토다안산대신경간세포증식화분화적영향。방법채용궤계취타결합200목강망려과적방법，종12．5 d 적곤명태서대뇌중분리 NSCs 병배양；용소단백（nestin）면역형광염색감정 NSC；가입불동농도다안산（theanine），MTS 법검측불동시간점활성，선정다안산최종가약농도（100μmol／L）；장 NSCs 분위량조：대조조、가약조（다안산농도위100μmol／L）；배양후진행 BrdU 염색화미관상관단백2（MAP2）、효질섬유산성단백（GFAP）면역형광염색；이용 Q-PCR 이급 Westen blotting 검측 MAP2화 GFAP 표체。결과100μmol／L 다안산처리조MTS 검측적흡광도치현저증고（P ＜0．05）；가약조 BrdU 양성세포비솔、MAP2양성세포비솔명현고우대조조（P ＜0．05），량조 GFAP 양성세포비솔무통계학차이；가약조 MAP2표체명현고우대조조（P ＜0．05），량조 GFAP표체무통계학차이。결론다안산가촉진 NSCs 증식급기향신경원방향분화。
Objective To explore the effects of theanine on the proliferation and differentiation of neural stem cells (NSCs).Methods NSCs were isolated from Kunming mouse embryonic at E12.5 by mechanical blowing combined with stencils of 200 mesh.Isolated NSCs were then identified with nestin immunofluorescent staining technique.Differ-ent concentrations of theanine were added into nutrient fluid,cell activity at different time was detected with MTS,and the final concentration of theanine (100 μmol/L)was determined.Then NSCs were divided into two groups:control group,theanine (100 μmol/L)treated group.The proliferation and differentiation of NSCs were determined with BrdU labeling and immunofluorescent staining of microtubule associated protein 2 (MAP2)and glial fibrillary acidic protein (GFAP).The Q-PCR and Western blotting were used to detect the expression of MAP2 and GFAP.Results The optical density of theanine (100 μmol/L)treated group was significantly increased in MTS (P <0.05).The ratio of BrdU-positive cells and MAP2-positive cells against total NSCs treated with theanine was significantly increased in com-parison with that in control group (P <0.05),but there was no noticeable difference in GFAP immunofluorescence between the two groups.The expression of MAP2 treated with theanine was significantly increased compared with that in the control group,there was no noticeable difference in GFAP expression between the two groups.Conclusion Theanine (100 μmol/L)can promote the proliferation of NSCs and its neuronal differentiation.