Research Papers:
Exploring the model of PC12 apoptosis induced by OGSD/R through in vitro experiments
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Abstract
Yanqing Sun1,*, Wei Zhu1,*, Shengyuan Zhou1, Zhiwei Wang1, Xiongsheng Chen1 and Lianshun Jia1
1Department of Spine, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
*These authors are considereds as co-first authors
Correspondence to:
Xiongsheng Chen, email: [email protected]
Lianshun Jia, email: [email protected]
Keywords: oxygen-glucose-serum deprivation/restoration, PC12, apoptosis, ischemia reperfusion
Received: March 01, 2017 Accepted: August 06, 2017 Published: October 04, 2017
ABSTRACT
Aims: To explicit cell apoptosis trend in PC12 oxygen-glucose-serum deprivation/restoration (OGSD/R) model and provide experimental bases for neural cell simulation in ischemia reperfusion injury in vitro.
Methods: OGSD/R model was constructed using the passage PC12 cells in vitro. The profile of cell apoptosis was estimated by DAPI staining, Annexin V-FITC and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay, as well as the levels of apoptosis-related proteins, including procaspase-3 and caspase-12.
Results: PC12 apoptosis was induced by OGSD and aggravated after restoration. CCK8 assay indicated that cell activity reached minimum after 1h of oxygen-glucose-serum restoration (OGR). DAPI staining suggested that apoptosis was the most serious after 1h of OGR, causing apoptotic cell nucleus pyknosis, particle spot formation, and fracture of cells with serious apoptosis forming pieces, and nucleus disintegration. The percentage of apoptotic cells exhibited increased trend after restoration, and reached the highest at 1h of OGR. Moreover, the expression of procaspase-3 and caspase-12 were extremely enhanced after OGD, especially 1h after OGR.
Conclusions: PC12 apoptosis is induced by OGSD and aggravated after restoration. The apoptosis of PC12 reaches the highest at 1h after OGR, which may provide experimental bases for spinal cord ischemia reperfusion injury treatment.
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