|
KMID : 1034820180140010079
|
|
Molecular & Cellular Toxicology
2018 Volume.14 No. 1 p.79 ~ p.86
|
|
|
Calreticulin protects the HT22 hippocampal neurons from injury induced by hypoxia
|
|
Wang Zhaoping
Yang Mei
Xue Jian
Chong Yanjun
|
|
|
Abstract
|
|
|
|
The purpose of this study was to evaluate the role of calreticulin (CRT) in the hypoxia-induced injury of HT22 cells through over-expression and knockdown of CRT. Cells were transfected with pcDNA3.1, pcDNA3.1 carrying full length CRT (pcDNA-CRT), small interfering RNA targeting CRT (si-CRT) or its negative control (siNC). Then, cell viability and apoptosis were respectively detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed to evaluate mRNA expression level of CRT. Western blotting was performed to determine the protein expression levels of CRT and apoptosis related proteins. Results showed CRT was successfully over-expressed by transfection of pcDNA-CRT (P<0.05) and silenced by transfection of si-CRT (P<0.05). CRT over-expression could promote the HT22 cell proliferation (P<0.01 or P<0.001) and inhibit hypoxia-induced apoptosis (P<0.05). Western blotting illustrated up-regulation of CRT increased the expression of Bcl-2 (P<0.01) but decreased the expressions of cytochrome c (P<0.01), active caspase 3 (P<0.001) and active caspase 9 (P<0.05) under hypoxia treatment. The effect of CRT knockdown was opposite to CRT over-expression. We speculated that CRT might regulate the expression of apoptosis related factors such as cytochrome-c, caspase 3, caspase 9 and Bcl-2, protecting the HT22 cells from hypoxia-induced injury. These results will provide the theory basis for the clinical treatment of cerebral hypoxia-induced injury.
|
|
|
KEYWORD
|
|
|
Calreticulin, Cell hypoxia, Caspase, Bcl-2, Cytochrome c
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|
|