解剖学报
解剖學報
해부학보
ACTA ANATOMICA SINICA
2014年
5期
627-632
,共6页
刘晓东%张夏南%郝宁%鞠前前%秦建兵%田美玲%金国华%吕广明
劉曉東%張夏南%郝寧%鞠前前%秦建兵%田美玲%金國華%呂廣明
류효동%장하남%학저%국전전%진건병%전미령%금국화%려엄명
海马%神经干细胞%雌二醇%增殖%雌激素受体%免疫荧光%大鼠
海馬%神經榦細胞%雌二醇%增殖%雌激素受體%免疫熒光%大鼠
해마%신경간세포%자이순%증식%자격소수체%면역형광%대서
Hippocampus%Neural stem cells%Estradiol%Proliferation%Estradiol receptor%Immunofluorescence%Rat
目的:观察雌二醇对大鼠海马神经干细胞增殖的影响及其作用机制。方法取20只孕17d的SD大鼠海马组织,进行神经干细胞的培养和增殖,添加不同浓度的雌二醇(E2)。通过5-溴脱氧尿嘧啶核苷(BrdU)免疫荧光、MTT方法检测神经干细胞的增殖;通过免疫荧光技术和巢蛋白( Nestin)双标观察神经干细胞球中雌激素受体ERα和ERβ的表达情况。结果 BrdU与MTT检测结果显示,随着雌二醇浓度从10-10 mol/L增加至10-8 mol/L,神经干细胞数量逐渐增加。雌二醇在10-8 mol/L时,细胞增殖数目最多而且细胞活力最好。随着雌二醇浓度进一步增加至10-6 mol/L,神经干细胞增殖能力逐渐下降。免疫荧光技术检测显示神经干细胞均表达ERα和ERβ两种受体。结论在一定浓度范围内,雌二醇能促进海马神经干细胞的增殖,并可能是通过ERα和ERβ介导促进神经干细胞增殖。
目的:觀察雌二醇對大鼠海馬神經榦細胞增殖的影響及其作用機製。方法取20隻孕17d的SD大鼠海馬組織,進行神經榦細胞的培養和增殖,添加不同濃度的雌二醇(E2)。通過5-溴脫氧尿嘧啶覈苷(BrdU)免疫熒光、MTT方法檢測神經榦細胞的增殖;通過免疫熒光技術和巢蛋白( Nestin)雙標觀察神經榦細胞毬中雌激素受體ERα和ERβ的錶達情況。結果 BrdU與MTT檢測結果顯示,隨著雌二醇濃度從10-10 mol/L增加至10-8 mol/L,神經榦細胞數量逐漸增加。雌二醇在10-8 mol/L時,細胞增殖數目最多而且細胞活力最好。隨著雌二醇濃度進一步增加至10-6 mol/L,神經榦細胞增殖能力逐漸下降。免疫熒光技術檢測顯示神經榦細胞均錶達ERα和ERβ兩種受體。結論在一定濃度範圍內,雌二醇能促進海馬神經榦細胞的增殖,併可能是通過ERα和ERβ介導促進神經榦細胞增殖。
목적:관찰자이순대대서해마신경간세포증식적영향급기작용궤제。방법취20지잉17d적SD대서해마조직,진행신경간세포적배양화증식,첨가불동농도적자이순(E2)。통과5-추탈양뇨밀정핵감(BrdU)면역형광、MTT방법검측신경간세포적증식;통과면역형광기술화소단백( Nestin)쌍표관찰신경간세포구중자격소수체ERα화ERβ적표체정황。결과 BrdU여MTT검측결과현시,수착자이순농도종10-10 mol/L증가지10-8 mol/L,신경간세포수량축점증가。자이순재10-8 mol/L시,세포증식수목최다이차세포활력최호。수착자이순농도진일보증가지10-6 mol/L,신경간세포증식능력축점하강。면역형광기술검측현시신경간세포균표체ERα화ERβ량충수체。결론재일정농도범위내,자이순능촉진해마신경간세포적증식,병가능시통과ERα화ERβ개도촉진신경간세포증식。
Objective The aim is to observe the role and mechanism of estradiol ( E2 ) on the proliferation of rat hippocampal neural stem cells ( NSCs ) .Methods Twenty hippocampi from embryonic 17-day ( E17 ) SD rats were dissociated and plated into culture flasks with NSCs specific medium containing different concentrations of estradiol .The proliferation and the vitality of NSCs were detected by immunofluorescence against BrdU and MTT assay .The expression of estrogen receptors ( ERαand ERβ) was measured by immunofluorescence staining combined with Nestin double labeling . Results BrdU and MTT assay results showed that the cell number increased when the concentration of estradiol increased from 10 -10 to 10 -8 mol/L.The number of cell proliferation and the viability of cells were best at the concentration of 10 -8 mol/L compared to the other groups .However, when the estradiol concentration was increased from 10-8 to 10 -6 mol/L, the cell proliferative capacity declined gradually .Double immunofluorescence labeling showed that the two types of estrogen receptors ( ERαand ERβ) were expressed in the cultured hippocampal NSCs .Conclusion Estradiol promotes the proliferation of hippocampal NSCs in a certain concentration range , and ERαand ERβmay be involved in the estradiol-induced proliferation .