低氧条件下PSB603干预对少突胶质前体细胞增殖和分化的影响

Effects of PSB603 intervention on proliferation and differentiation of oligodendrocyte precursor cells under hypoxia

目的:探讨在低氧条件下,腺苷受体A2b(A2bAR)拮抗剂PSB603对少突胶质前体细胞(OPC)增殖和分化的影响。方法:剥离新生2 d SD大鼠的脑皮层,并原代培养其OPC,随机分为对照组、PSB603组、2%O_2组和实验组,实验组在2%O_2干预的基础上再给予PSB603处理,4 d后用BrdU掺入技术和免疫细胞化学方法检测BrdU阳性细胞百分数和CNPase阳性细胞百分数的变化,并用qRT-PCR技术检测各组细胞P21cip1、P27kip1 mRNA含量的变化。结果:与对照组相比较,2%O_2组的BrdU阳性细胞百分数显著上升(P<0.01),P21、P27 mRNA含量均显著下调(P<0.01),各组间CNPase阳性细胞百分数无明显变化;实验组与2%O_2组间各实验差异均无统计学意义(P>0.05)。结论:低氧短时间干预能够促进OPC细胞的增殖,P21cip1和P27kip1参与其调控,A2b AR未改变低氧所诱导的OPC增殖。

Objective：To investigate the effect of adenosine receptor A2b （A2bAR） antagonist PSB603 on the proliferation and differentiation of oligodendrocyte precursor cells （OPC） under hypoxia.Methods：The cerebral cortex of neonatal SD rats was dissected and primarily cultured with OPC which were randomly divided into control group,PSB603 group,2% O2 group and experimental group.The experimental group was treated with PSB603 after 2% O2 intervention.The percentage of BrdU-positive cells and CNPase-positive cells were detected by BrdU incorporation and immunocytochemistry.The contents of P21cip1 and P27kip1 mRNA levels were detected by qRT-PCR.Results：Compared with the control group,the percentage of BrdU positive cells in 2% O2 group increased significantly （P 〈 0.01）,the mRNA levels of P21 and P27 decreased significantly （P 〈0.01）;The percentage of CNPase positive cells did not change significantly among each group;There was no significant difference of the percentage of BrdU positive cells or CNPase positive cells or the mRNA levels of P21 and P27 between the 2% O2 group and the control group （P 〉 0.05）.Conclusion：Hypoxia short-term intervention can promote the proliferation of OPC cells,which is involved into P21 cip1 and P27kip1.A2bAR does not change the OPC-induced proliferation induced by hypoxia.