Expression and role of Pim1 in cultured cortical neurons with oxygen-glucose deprivation/reoxygen injury

LIU Jun-Yan; WANG Ke-Xuan; HUANG Ling-Yi; WAN Bin; ZHAO Guo-Ying; ZHAO Feng-Yan

doi:10.7499/j.issn.1008-8830.1911045

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Chinese Journal of Contemporary Pediatrics ›› 2020, Vol. 22 ›› Issue (5) : 512-518. DOI: 10.7499/j.issn.1008-8830.1911045

EXPERIMENTAL RESEARCH

Expression and role of Pim1 in cultured cortical neurons with oxygen-glucose deprivation/reoxygen injury

LIU Jun-Yan¹, WANG Ke-Xuan¹, HUANG Ling-Yi², WAN Bin¹, ZHAO Guo-Ying¹, ZHAO Feng-Yan³

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Abstract

Objective To study the expression and effect of Pim1 in primary cortical neurons after hypoxic-ischemic injury. Methods Cortical neurons were isolated from 1-day-old C57BL/6 mice and cultured in neurobasal medium. On the 8th day of neuron culture, cells were subjected to oxygen-glucose deprivation/reoxygen (OGD/R) treatment to mimic in vivo hypoxic injury of neurons. Briefly, medium were changed to DMEM medium, and cells were cultured in 1% O₂ for 3 hours and then changed back to normal medium and conditions. Cells were collected at 0 hour, 6 hours, 12 hours and 24 hours after OGD/R. Primary neurons were transfected with Pim1 overexpression plasmid or mock plasmid, and then were exposed to normal conditions or OGD/R treatment. They were named as Pim1 group, control group, OGD/R group and OGD/R+Pim1 group respectively. Real-time PCR was used to detect Pim1 mRNA expression. Western blot was used to detect the protein expression of Pim1 and apoptotic related protein cleaved caspase 3 (CC3). TUNEL staining was used to detect cell apoptosis. Results Real-time PCR and Western blot results showed that Pim1 mRNA and protein were significantly decreased in neurons after OGD/R. They began to decrease at 0 hour after OGD/R, reached to the lowest at 12 hours after OGD/R, and remained at a lower level at 24 hours after OGD/R (P < 0.01). Overexpression of Pim1 significantly upregulated the protein level of Pim1. Under OGD/R conditions, the CC3 expression and the apoptosis rate in cells of the Pim1 group were significantly lower than in un-transfected cells (P < 0.01). Conclusions Hypoxic-ischemic injury may decrease Pim1 expression in neurons. Overexpressed Pim1 may inhibit apoptosis induced by OGD/R.

Key words

Brain injury / Apoptosis / Hypoxia ischemia / Pim1 / Neuron

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LIU Jun-Yan, WANG Ke-Xuan, HUANG Ling-Yi, WAN Bin, ZHAO Guo-Ying, ZHAO Feng-Yan. Expression and role of Pim1 in cultured cortical neurons with oxygen-glucose deprivation/reoxygen injury[J]. Chinese Journal of Contemporary Pediatrics. 2020, 22(5): 512-518 https://doi.org/10.7499/j.issn.1008-8830.1911045

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