中华物理医学与康复杂志
中華物理醫學與康複雜誌
중화물리의학여강복잡지
CHINESE JOURNAL OF PHYSICAL MEDICINE AND REHABILITATION
2015年
5期
327-331
,共5页
柳华%韩肖华%陈红%黄晓琳
柳華%韓肖華%陳紅%黃曉琳
류화%한초화%진홍%황효림
重复磁刺激%大鼠%神经干细胞%细胞增殖
重複磁刺激%大鼠%神經榦細胞%細胞增殖
중복자자격%대서%신경간세포%세포증식
Repetitive magnetic stimulation%Rats%Neural stem cells%Cell proliferation
目的 探讨重复磁刺激(rMS)对体外培养大鼠神经干细胞(NSCs)增殖的作用机制.方法 取新生3天内的Sprague-Dawley(SD)大鼠乳鼠双侧海马组织培养NSCs,通过cck-8试剂盒检测第2代NSCs的OD值绘制生长曲线图.再将第2代NSCs分为空白对照组和rMS组,rMS组刺激参数为频率10 Hz,50％最大输出强度,每天200个脉冲,连续刺激3d.于rMS组最后1次干预1h后收集2组细胞,采用cck-8试剂盒检测其细胞增殖效应,同时用免疫印迹法检测c-fos蛋白和环磷酸腺苷反应元件结合蛋白(p-CREB)的表达量.结果 第2代神经球经巢蛋白(nestin)免疫荧光染色证实为NSCs,生长曲线提示培养第3天时NSCs活性最佳.rMS干预后,rMS组cck-8的OD值为(0.309±0.043),与空白对照组的(0.256±0.043)比较,差异有统计学意义(P ＜0.05);rMS干预后,rMS组的c-fos和p-CREB蛋白相对表达量分别与空白对照组比较,差异均有统计学意义(P＜0.01).结论 频率10 Hz的rMS可促进NSCs的增殖,其作用机制可能与p-CREB和c-fos蛋白表达的增加有关.
目的 探討重複磁刺激(rMS)對體外培養大鼠神經榦細胞(NSCs)增殖的作用機製.方法 取新生3天內的Sprague-Dawley(SD)大鼠乳鼠雙側海馬組織培養NSCs,通過cck-8試劑盒檢測第2代NSCs的OD值繪製生長麯線圖.再將第2代NSCs分為空白對照組和rMS組,rMS組刺激參數為頻率10 Hz,50％最大輸齣彊度,每天200箇脈遲,連續刺激3d.于rMS組最後1次榦預1h後收集2組細胞,採用cck-8試劑盒檢測其細胞增殖效應,同時用免疫印跡法檢測c-fos蛋白和環燐痠腺苷反應元件結閤蛋白(p-CREB)的錶達量.結果 第2代神經毬經巢蛋白(nestin)免疫熒光染色證實為NSCs,生長麯線提示培養第3天時NSCs活性最佳.rMS榦預後,rMS組cck-8的OD值為(0.309±0.043),與空白對照組的(0.256±0.043)比較,差異有統計學意義(P ＜0.05);rMS榦預後,rMS組的c-fos和p-CREB蛋白相對錶達量分彆與空白對照組比較,差異均有統計學意義(P＜0.01).結論 頻率10 Hz的rMS可促進NSCs的增殖,其作用機製可能與p-CREB和c-fos蛋白錶達的增加有關.
목적 탐토중복자자격(rMS)대체외배양대서신경간세포(NSCs)증식적작용궤제.방법 취신생3천내적Sprague-Dawley(SD)대서유서쌍측해마조직배양NSCs,통과cck-8시제합검측제2대NSCs적OD치회제생장곡선도.재장제2대NSCs분위공백대조조화rMS조,rMS조자격삼수위빈솔10 Hz,50％최대수출강도,매천200개맥충,련속자격3d.우rMS조최후1차간예1h후수집2조세포,채용cck-8시제합검측기세포증식효응,동시용면역인적법검측c-fos단백화배린산선감반응원건결합단백(p-CREB)적표체량.결과 제2대신경구경소단백(nestin)면역형광염색증실위NSCs,생장곡선제시배양제3천시NSCs활성최가.rMS간예후,rMS조cck-8적OD치위(0.309±0.043),여공백대조조적(0.256±0.043)비교,차이유통계학의의(P ＜0.05);rMS간예후,rMS조적c-fos화p-CREB단백상대표체량분별여공백대조조비교,차이균유통계학의의(P＜0.01).결론 빈솔10 Hz적rMS가촉진NSCs적증식,기작용궤제가능여p-CREB화c-fos단백표체적증가유관.
Objective To study the mechanism of rats' neural stem cells (NSCs) proliferation in vitro after repetitive magnetic stimulation (rMS).Metbods The bilateral hippocampus of neonatal Sprague-Dawley rats (＜3 d) was taken out to culture NSCs in vitro.The OD value was evaluated with Cell Counting Kit-8 (cck-8) and cell growth curve was generated.The NSCs cultured were divided into a control group and an rMS group.rMS (3 days,once per day) was applied on p2 NSCs at 10 Hz,50％ machine output and 200 pulses per day.One hour after the last rMS,the cck-8 was used to test the cell proliferation,and the western blotting was applied to detect the protein expression of c-fos and p-CREB.Results The nestin fluorescent staining of p2 neurospheres was proved to be neural stem cells.The growth curve indicated that their viability reached the peak on the third day.The OD value in the rMS group (0.309 ± 0.043) showed a significant difference (P ＜ 0.05) after rMS compared with the control group (0.256± 0.043).So did the c-fos and p-CREB protein expression between the two groups (P ＜ 0.01).Conclusion The rMS at 10 Hz can promote rats' NSCs proliferation in vitro,which may be related to the increased expression of p-CREB and c-fos after rMS.
