ABSTRACT
Objective:
To investigate the expression of
zinc finger protein 580 (ZNF580) in
oxygen-
glucose deprivation (OGD) model of SH-SY5Y
cell line and its overexpression on the
apoptosis of hypoxic-ischemic
neurons and the possible mechanism.
Methods:
The study was divided into two parts (1)
Human neuroblastoma SH-SY5Y
cell line was cultured and divided into the model group and
control group. The model group was incubated at 37 ℃ for 6 h in a three-gas
incubator of 95% N 2, 5% CO 2, and 0.1% O 2 to establish OGD model, and
proteins were extracted at 6, 12, and 24 h after OGD. The expression of ZNF580 was quantified by
Western blot. (2) Effects of ZNF580 overexpressed with
lentivirus transfection on the
apoptosis and cleaved
caspase-3 expression
Cells were collected from the
control group and model group 24 h after OGD. Overexpressed ZNF580
cells were constructed by
lentivirus transfection as the overexpression group and then treated with OGD.
Flow cytometry was used to detect the
apoptosis rate in the three groups and
Western blot was used to detect the expression of cleaved
caspase-3. Two independent sample t-test, one-way
variance analysis, and
LSD- t for pairwise comparison were used for
statistical analysis.
Results:
(1) ZNF580 expression was significantly increased at 6, 12, and 24 h after OGD compared with the
control group (1.36±0.05, 2.12±0.07, 1.69±0.05 vs 1.00,
LSD- t=9.20, 28.26, and 19.21, all P<0.001). (2)
Apoptosis rates of the control, model, and overexpression groups were (1.07±0.56)%, (21.51±1.65)%, and (3.42±0.93)%, respectively, and relative expression levels of cleaved
caspase-3 were 1.00, 2.47±0.59, and 1.70±0.25, respectively. Compared with the
control group,
apoptosis rate and cleaved
caspase-3 relative expression level were significantly increased in the model group (
LSD- t=21.98 and 8.17, both P=0.001), while the two figures were significantly decreased in the overexpression group when compared with the model group (
LSD- t=19.45, P=0.001;
LSD- t=4.28, P=0.005).
Conclusion:
Hypoxia and
ischemia could
lead to the overexpression of ZNF580, which may reduce the
apoptosis of hypoxic-ischemic
neurons by inhibiting the expression of cleaved
caspase-3 and affecting its enzymatic activation.