Expression and role of Pim1 in cultured cortical neurons with oxygen-glucose deprivation/reoxygen injury.
- Author:
Jun-Yan LIU
1
;
Ke-Xuan WANG
;
Ling-Yi HUANG
;
Bin WAN
;
Guo-Ying ZHAO
;
Feng-Yan ZHAO
Author Information
1. Department of Neonatology, Binzhou Medical University Hospital, Binzhou, Shandong 256600, China. zhaofengyan621@163.com.
- Publication Type:Journal Article
- MeSH:
Animals;
Glucose;
Mice;
Mice, Inbred C57BL;
Neurons;
Oxygen;
Proto-Oncogene Proteins c-pim-1;
Rats, Sprague-Dawley
- From:
Chinese Journal of Contemporary Pediatrics
2020;22(5):512-518
- CountryChina
- Language:Chinese
-
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% O 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.
