天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2013年
8期
786-788
,共3页
崔猛%冯世庆%范宁建%贾军%周先虎
崔猛%馮世慶%範寧建%賈軍%週先虎
최맹%풍세경%범저건%가군%주선호
黄芩甙%信号传导%转录,遗传%转录因子%白血病%干细胞%神经系统%细胞分化
黃芩甙%信號傳導%轉錄,遺傳%轉錄因子%白血病%榦細胞%神經繫統%細胞分化
황금대%신호전도%전록,유전%전록인자%백혈병%간세포%신경계통%세포분화
Baicalin%signal transduction%transcription,genetic%transcription factors%leukemia%stem cells%ner-vous system%cell differentiation
目的观察黄芩苷促进神经干细胞（NSCs）体外向神经元分化过程中信号传导子与转录激活子(STAT)的磷酸化蛋白表达水平。方法从孕14～15 d的SD大鼠胚胎大脑皮质中分离NSCs，体外培养传代，实验用第3代NSCs。随机分为对照组，低、中、高黄芩苷组（分别含黄芩苷7.5、15、30μmol/L），白血病抑制因子（LIF）+碱性成纤维细胞生长因子（bFGF）组和黄芩苷+LIF+bFGF组。培养6 d后，细胞免疫荧光化学染色法观察各组中微管相关蛋白2(MAP-2)、胶质纤维酸性蛋白（GFAP）表达水平。培养2 h与6 d后，用Western blot方法观察各组中STAT3蛋白磷酸化水平。结果黄芩苷可使NSCs中MAP-2表达增加，GFAP表达减弱；LIF+bFGF可促进NSCs中GFAP表达增加；黄芩苷可抑制LIF+bFGF引起的NSCs中GFAP的表达增加。黄芩苷可下调NSCs中STAT3蛋白磷酸化水平；LIF+bFGF可上调NSCs中STAT3蛋白磷酸化水平；黄芩苷可抑制LIF+bFGF引起的NSCs中STAT3蛋白磷酸化水平的上调（P＜0.05）。结论黄芩苷可诱导NSCs向神经元分化并抑制其向星形胶质细胞分化，该作用可能经下调NSCs中STAT3的磷酸化水平来实现。
目的觀察黃芩苷促進神經榦細胞（NSCs）體外嚮神經元分化過程中信號傳導子與轉錄激活子(STAT)的燐痠化蛋白錶達水平。方法從孕14～15 d的SD大鼠胚胎大腦皮質中分離NSCs，體外培養傳代，實驗用第3代NSCs。隨機分為對照組，低、中、高黃芩苷組（分彆含黃芩苷7.5、15、30μmol/L），白血病抑製因子（LIF）+堿性成纖維細胞生長因子（bFGF）組和黃芩苷+LIF+bFGF組。培養6 d後，細胞免疫熒光化學染色法觀察各組中微管相關蛋白2(MAP-2)、膠質纖維痠性蛋白（GFAP）錶達水平。培養2 h與6 d後，用Western blot方法觀察各組中STAT3蛋白燐痠化水平。結果黃芩苷可使NSCs中MAP-2錶達增加，GFAP錶達減弱；LIF+bFGF可促進NSCs中GFAP錶達增加；黃芩苷可抑製LIF+bFGF引起的NSCs中GFAP的錶達增加。黃芩苷可下調NSCs中STAT3蛋白燐痠化水平；LIF+bFGF可上調NSCs中STAT3蛋白燐痠化水平；黃芩苷可抑製LIF+bFGF引起的NSCs中STAT3蛋白燐痠化水平的上調（P＜0.05）。結論黃芩苷可誘導NSCs嚮神經元分化併抑製其嚮星形膠質細胞分化，該作用可能經下調NSCs中STAT3的燐痠化水平來實現。
목적관찰황금감촉진신경간세포（NSCs）체외향신경원분화과정중신호전도자여전록격활자(STAT)적린산화단백표체수평。방법종잉14～15 d적SD대서배태대뇌피질중분리NSCs，체외배양전대，실험용제3대NSCs。수궤분위대조조，저、중、고황금감조（분별함황금감7.5、15、30μmol/L），백혈병억제인자（LIF）+감성성섬유세포생장인자（bFGF）조화황금감+LIF+bFGF조。배양6 d후，세포면역형광화학염색법관찰각조중미관상관단백2(MAP-2)、효질섬유산성단백（GFAP）표체수평。배양2 h여6 d후，용Western blot방법관찰각조중STAT3단백린산화수평。결과황금감가사NSCs중MAP-2표체증가，GFAP표체감약；LIF+bFGF가촉진NSCs중GFAP표체증가；황금감가억제LIF+bFGF인기적NSCs중GFAP적표체증가。황금감가하조NSCs중STAT3단백린산화수평；LIF+bFGF가상조NSCs중STAT3단백린산화수평；황금감가억제LIF+bFGF인기적NSCs중STAT3단백린산화수평적상조（P＜0.05）。결론황금감가유도NSCs향신경원분화병억제기향성형효질세포분화，해작용가능경하조NSCs중STAT3적린산화수평래실현。
Objective To observe the role of baicalin on the expression of phosphorylated protein of signal transduc-ers and activators of transcription signaling proteins (STATs) during the process that neural stem cells (NSCs) differentiating into neurons. Methods NSCs were isolated from the embryonic cerebral cortex of the 14-15-day pregnant SD rats, which were cultured and passaged in vitro. The 3rd generation of NSCs was used in the experiment. NSCs were randomized into nat-ural differentiation control group, three different doses of baicalin groups (7.5μmol/L, 15μmol/L and 30μmol/L), leukemia inhibitory factor (LIF)+basic fibroblast growth factor (bFGF) group and baicalin+LIF+bFGF group. After 6 d culture in vi-tro, the immunohistochemical method was used to observe the expressions of microtubule-associated protein 2(MAP-2) and glial fibrillary acidic protein (GFAP) in different groups. The expression levels of phosphorylation protein of STAT 3 in NSCs were detected by Western blotting method after 2 h and 6 d of culture. Results The expression of MAP-2 in NSCs was in-creased by baicalin, but the expression of GFAP in NSCs decreased. The expression of GFAP in NSCs was enhanced in LIF+bFGF group, which was inhibited by baicalin+LIF+bFGF. The phosphorylation level of STAT3 in NSCs was downregulat-ed by baicalin, but the phosphorylation level of STAT3 was upregulated in LIF+bFGF group. The upregulated phosphoryla-tion level of STAT3 was inhibited in baicalin+LIF+bFGF group(P<0.05). Conclusion Baicalin can induce NSCs to dif-ferentiate into neurons, which may be caused by the downregulation of the phosphorylation level of STAT3 in NSCs.
