缺氧调节基因/产物参与CoCl_2预处理对分化SH-SY5Y细胞缺氧损伤的保护

Roles of hypoxia regulated genes/protein in neuroprotection against hypoxia injury in differentiated SH-SY5Y cells preconditioned with CoCl_2

目的探讨缺氧调节基因/产物在CoCl2化学缺氧预处理抵抗神经型细胞缺氧损伤中的作用。方法分化的SH-SY5Y细胞分为对照组、化学缺氧预处理组(细胞先用50μmol/LCoCl2预处理3h,换液后常氧培养1h,然后在2%的低氧孵箱内缺氧28h)、缺氧组(2%的低氧孵箱内缺氧28h)。RT-PCR测VEGF、GLUT-1、EPO、LDH-A的mRNA表达。通过应用重组人VEGF纯品,进一步确定VEGF在化学缺氧预处理组中的保护作用。结果化学缺氧预处理组细胞GLUT-1、EPO mRNA表达显著高于缺氧组(P<0·05),VEGF mRNA表达显著高于缺氧组(P<0·01),LDH-A mRNA表达在两组之间没有显著性差异(P>0·05)。MTT细胞活力测定显示,重组人VEGF可模拟预处理组的保护作用。结论CoCl化学缺氧预处理很可能通过促进VEGF、GLUT-1、EPO等缺氧调节基因/产物表达,发挥其神经保护作用。

Objective To investigate the roles of hypoxia regulated genes/protein in chemical hypoxia preconditioning in differentiated SH-SY5Y cells. Methods Differentiated SH-SY5Y cells were randomly divided into control group, chemical hypoxic preconditioning group （50 μmol/L CoCl2 preconditioning for 3 h, normal culture for 1 h, then in 2% O2 for 28 h） and hypoxia group （ in 2% O2 for 28 h）. RT-PCR was used to examine the mRNA expressions of VEGF, GLUT-1, EPO, LDH-A. Further evaluation of the potential neuroprotective effect of VEGF was done by investigating the effect of recombinant human VEGF on subsequent hypoxia injury. Results The mRNA levels of GLUT-1, EPO increased in the preconditioning group as compared with those in the hypoxia group （P 〈 0. 05 ）, so did VEGF mRNA expression （P 〈 0. 01 ）, but the LDH-A mRNA expression had no apparent difference between the two groups. MTT assay showed the effect of precondition could be simulated by recombinant human VEGF. Conclusion The upregulated expression of VEGF, GLUT-1, EPO genes/ protein level might be involved in the neuroprotection of chemical hypoxia preconditioning.