脑溢安对大鼠缺氧神经干细胞保护作用

Protective effect of naoyian on neural stem cells in rats with anoxia

目的:观察脑溢安对大鼠神经干细胞缺氧损伤后天冬氨酸特异酶切的半胱氨酸蛋白酶-3(activatedcysteinylaspartatespecificprotease-3,Caspase-3)活化及细胞凋亡的影响,探讨该药抗脑损伤的作用机制。方法:用分离、培养的神经干细胞(neuralstemcellsNSC)造成缺氧模型,以正常大鼠血清、含脑溢安的大鼠血清对缺氧后的神经干细胞进行干预,用脑溢安血清、正常血清、无血清干预缺氧的神经干细胞,以及未受缺氧损伤的正常神经干细胞进行实验,检测神经干细胞缺氧损伤后Caspase-3的活化情况并用末端脱氧核苷酸转移酶介导dUTP缺口末端标记法犤terminal(TdT)-mediateddUTP-biotinnickendlabeling,TUNEL犦检测细胞凋亡情况。结果:正常血清缺氧神经干细胞组、无血清缺氧神经干细胞组有较多TUNEL阳性细胞,脑溢安血清缺氧神经干细胞组TUNEL阳性细胞数较少,而未缺氧的神经干细胞没有表达,脑溢安血清缺氧神经干细胞组与正常血清缺氧神经干细胞组、无血清缺氧神经干细胞组比较TUNEL细胞阳性细胞减少,差异有显著性意义(t=6.5826,6.6245,P<0.05)。脑溢安血清缺氧神经干细胞组与正常血清缺氧神经干细胞组及无血清缺氧神经干细胞组比较活化Caspase-3光密度比值减少,差异有显著性意义(t=4.2653,5.0131,P<0.05)。无血清。

AIM:To study the effect of naoyian on the expression of activated cysteinyl as partate specific protease-3(activated Caspase-3) of neural stem cells(NSCs) in fuced by anoxia and apoptosis and to explore its mechanism on brain protection. METHODS:NSCs taken from rats and cultured in solution were used to establish a noxia model. Normal rat serum and rat serum containing naoyian were used to inte rvene NSCs after anoxia.The expression of activated Capase-3 in anoxic NSCs and normal NSCs were detected while the apoptosis was examined with terminal(TdT)- mediated dUTP-biotin nick end labeling(TUNEL). RESULTS:TUNEL positive cells expressed more in the normal serum group with NSC s after anoxia and the serum-free group with NSCs after anoxia,while less in NY A serum group with NSCs after anoxia and none in the NSCs without anoxia.The TUN EL positive cells expressed less in naoyian serum group with NSCs after anoxia t han those in the normal serum group with NSCs after anoxia and the serum-free g roup with NSCs after anoxia, and the differences were significant(t=6.582 6,6.62 4 5,P< 0.05).The absorbance of the activited Caspase-3 in NYA serum group was l ower than that in the normal serum group and serum-free group separately. The d ifferences were significant(t=4.265 3,5.013 1,P< 0.05).There were no significant differences between the serum-free group and the normal serum group with NSCs after anoxia(t=0.519 0, 0.265 7,P >0.05). CONCLUSION:Serum containing naoyian can intervene NSC injured by anoxia,decrea se the activity of Caspase-3 and decrease the apoptosis of NSCs after anoxia,an d therefore protect brain.