大鼠腦中風(fēng)后移植GDNF基因修飾的神經(jīng)干細(xì)胞激活p44-42MAPK與AKT細(xì)胞信號(hào)通路.pdf

1、目的:研究膠質(zhì)源性神經(jīng)營(yíng)養(yǎng)因子(glial cell line-derived neuralfactor,GDNF)基因修飾的神經(jīng)干細(xì)胞(neural stem cells,NSCs)移植對(duì)大鼠腦缺血后神經(jīng)保護(hù)作用機(jī)制是否與促絲裂原活化蛋白激酶(mitogen—activated protein kinase,p44/42MAPK)及其特異性抑制劑絲裂原活化蛋白激酶磷酸酶-1(Mitogen-activated protein kina

2、sephosphatase-1,MKP-1)和磷酸肌醇3激酶/絲氨酸-蘇氨酸酶(phosphatidylinositol-3kinase/serine-theonine kinase,PI3K/AKT)信號(hào)通路激活有關(guān)。方法:建立大鼠暫時(shí)性大腦中動(dòng)脈梗塞模型(middlecerebral artery occlusion,MCAO),再灌注3天,分別移植神經(jīng)干細(xì)胞(neural stem cells,NSCs)、GDNF基因轉(zhuǎn)染的NSC

3、s(GDNF/NSCs)和給予生理鹽水(對(duì)照組,NS)。每組根據(jù)再灌注時(shí)間不同分為1w、2w、3w、5w、7w5個(gè)亞組,每個(gè)時(shí)間點(diǎn)7只大鼠:4只用來(lái)做凋亡細(xì)胞的TUNEL染色,檢測(cè)神經(jīng)細(xì)胞的凋亡數(shù)；3只用來(lái)做蛋白免疫印跡(Western—blotting),檢測(cè)MKP-1和MAPK、AKT的磷酸化程度(pMAPK/MAPK、pAKT/AKT)。結(jié)果:1.GDNF/NSCs組、NSCs組和NS組的神經(jīng)細(xì)胞凋亡的數(shù)目在1w、2w、3w、5w

4、、7w各時(shí)間點(diǎn)均逐漸降低；GDNF/NSCs組和NSCs組神經(jīng)細(xì)胞凋亡數(shù)目顯著低于NS組(P=0.000 P=0.01)；GDNF/NSCs組神經(jīng)細(xì)胞凋亡數(shù)目顯著低于NSCs組(P=0.005)。2.GDNF/NSCs組、NSCs組和NS組pMAPK/MAPK在1w、2w、3w、5w、7w各時(shí)間點(diǎn)均逐漸降低；在各時(shí)間點(diǎn),GDNF/NSCs組和NSCs組pMAPK/MAPK顯著高于NS組(P=0.000P=0.000)；GDNF/NSCs

5、組pMAPK/MAPK顯著高于NSCs組(P=0.000)。3.GDNF/NSCs組、NSCs組和NS組MKP-1于2w達(dá)到高峰后在3w、5w、7w逐漸降低；NS組NSCs組MKP-1明顯高于GDNF/NSCs組(P=0.000 P=0.001)；NS組MK-1明顯高于NSCs組(P=0.005)。4.GDNF/NSCs組、NSCs組和NS組pAKT/AKT在1w、2w、3w、5w、7w各時(shí)間點(diǎn)均逐漸降低；再灌注各時(shí)間點(diǎn)GDNF/NSC