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
2014年
4期
246-249
,共4页
尹德龙%靳国青%程鹏%胡汉生%范震波%黄晖%郭风劲%陈安民
尹德龍%靳國青%程鵬%鬍漢生%範震波%黃暉%郭風勁%陳安民
윤덕룡%근국청%정붕%호한생%범진파%황휘%곽풍경%진안민
表皮干细胞%低强度电磁场%分化
錶皮榦細胞%低彊度電磁場%分化
표피간세포%저강도전자장%분화
Epidermis stem cells%Electromagnetic fields%Differentiation
目的 观察不同频率低强度电磁场对表皮干细胞(EpSCs)分化的影响,并从基因水平初步探讨其调控机制.方法 取SD大鼠乳鼠背部皮肤组织,通过体外分离培养获取第2代EpSCs,将其分为3个磁刺激组及对照组,各磁刺激组细胞分别给予15,50及75 Hz,5 mT电磁场干预,对照组细胞未给予电磁场刺激.于实验进行10 d后对各组细胞进行免疫荧光检测以了解细胞分化情况,同时采用RT-PCR技术检测各组细胞c-myc、β-连环蛋白(β-catenin) mRNA基因表达,应用免疫组化方法检测各组细胞角蛋白18 (CK18)表达情况.结果 不同频率低强度电磁场刺激均能促进EpSCs分化,各磁刺激组细胞分化相关角蛋白(即CK18)表达均为阳性,并以50 Hz磁刺激组CK18阳性表达尤为显著.各磁刺激组c-myc mRNA表达(15,50及75 Hz磁刺激组c-myc mRNA吸光度值分别为27324,37037及24790)均较对照组增强(对照组c-myc mRNA吸光度值为19037) (P ＜0.01),且以50 Hz磁刺激组上调幅度较显著(P＜0.05).β-catenin mRNA表达(15,50及75 Hz磁刺激组β-catenin mRNA吸光度值分别为23199,22385及15688)则较对照组(对照组β-catenin mRNA吸光度值为30591)减低(P＜0.05).结论 15,50及75 Hz,5 mT电磁场刺激均能促进EpSCs分化,并以50 Hz,5 mT电磁场对EpSCs的促分化作用较显著,其调控机制可能与增强细胞c-myc表达、抑制β-catenin表达有关.
目的 觀察不同頻率低彊度電磁場對錶皮榦細胞(EpSCs)分化的影響,併從基因水平初步探討其調控機製.方法 取SD大鼠乳鼠揹部皮膚組織,通過體外分離培養穫取第2代EpSCs,將其分為3箇磁刺激組及對照組,各磁刺激組細胞分彆給予15,50及75 Hz,5 mT電磁場榦預,對照組細胞未給予電磁場刺激.于實驗進行10 d後對各組細胞進行免疫熒光檢測以瞭解細胞分化情況,同時採用RT-PCR技術檢測各組細胞c-myc、β-連環蛋白(β-catenin) mRNA基因錶達,應用免疫組化方法檢測各組細胞角蛋白18 (CK18)錶達情況.結果 不同頻率低彊度電磁場刺激均能促進EpSCs分化,各磁刺激組細胞分化相關角蛋白(即CK18)錶達均為暘性,併以50 Hz磁刺激組CK18暘性錶達尤為顯著.各磁刺激組c-myc mRNA錶達(15,50及75 Hz磁刺激組c-myc mRNA吸光度值分彆為27324,37037及24790)均較對照組增彊(對照組c-myc mRNA吸光度值為19037) (P ＜0.01),且以50 Hz磁刺激組上調幅度較顯著(P＜0.05).β-catenin mRNA錶達(15,50及75 Hz磁刺激組β-catenin mRNA吸光度值分彆為23199,22385及15688)則較對照組(對照組β-catenin mRNA吸光度值為30591)減低(P＜0.05).結論 15,50及75 Hz,5 mT電磁場刺激均能促進EpSCs分化,併以50 Hz,5 mT電磁場對EpSCs的促分化作用較顯著,其調控機製可能與增彊細胞c-myc錶達、抑製β-catenin錶達有關.
목적 관찰불동빈솔저강도전자장대표피간세포(EpSCs)분화적영향,병종기인수평초보탐토기조공궤제.방법 취SD대서유서배부피부조직,통과체외분리배양획취제2대EpSCs,장기분위3개자자격조급대조조,각자자격조세포분별급여15,50급75 Hz,5 mT전자장간예,대조조세포미급여전자장자격.우실험진행10 d후대각조세포진행면역형광검측이료해세포분화정황,동시채용RT-PCR기술검측각조세포c-myc、β-련배단백(β-catenin) mRNA기인표체,응용면역조화방법검측각조세포각단백18 (CK18)표체정황.결과 불동빈솔저강도전자장자격균능촉진EpSCs분화,각자자격조세포분화상관각단백(즉CK18)표체균위양성,병이50 Hz자자격조CK18양성표체우위현저.각자자격조c-myc mRNA표체(15,50급75 Hz자자격조c-myc mRNA흡광도치분별위27324,37037급24790)균교대조조증강(대조조c-myc mRNA흡광도치위19037) (P ＜0.01),차이50 Hz자자격조상조폭도교현저(P＜0.05).β-catenin mRNA표체(15,50급75 Hz자자격조β-catenin mRNA흡광도치분별위23199,22385급15688)칙교대조조(대조조β-catenin mRNA흡광도치위30591)감저(P＜0.05).결론 15,50급75 Hz,5 mT전자장자격균능촉진EpSCs분화,병이50 Hz,5 mT전자장대EpSCs적촉분화작용교현저,기조공궤제가능여증강세포c-myc표체、억제β-catenin표체유관.
Objective To observe the effect of a low-intensity pulsed electromagnetic field (LIPEMF) on the differentiation of epidermal stem cells (EpSCs),and to explain the regulation of any observed effect on the gene level.Methods EpSCs were isolated from the back skin of neonatal Sprague-Dawley rats,and divided into three experimental groups and a control group.The three experimental aliquots received 5 mT magnetic field irradiation pulsed at 15 Hz,50 Hz or 75 Hz.After 10 days of continuous stimulation,immunofluorescence was used to observe any differentiation,RT-PCRs tested for c-myc and β-catenin mRNA,and immunohistochemical methods were applied to test for cytokeratin 18 (CK18).Results The EpSCs of the three stimulation groups all differentiated,and differentiation-related CK 18 showed significant expression in the 50 Hz group.Expression of c-myc mRNA increased significantly in all three experimental samples compared with the controls.Expression of β-catenin mRNA decreased significantly in all three samples compared with the controls.Conclusion LIPEMFs can induce the differentiation of EpSCs.Irradiation at 50 Hz and 5 mT is the most effective.The effect may be related to enhanced expression of c-myc and inhibited expression of β-catenin.
