Ferrostatin-1 delay D-gal induced cardiomyocyte senescence by inhibiting ferroptosis

doi:10.11958/20220818

Abstract

Abstract:

Objective To investigate the existence of ferroptosis in cardiomyocyte senescence induced by D-galactose (D-gal) and whether ferroptosis inhibitor ferrostatin-1 (Fer-1) can delay cardiomyocyte senescence. Methods Cardiomyocyte injury was induced by D-gal (0, 5, 10, 20, 40, 80 and 100 g/L) to simulate cardiac aging model. After cells were treated with different concentrations of D-gal for 24 hours, the cell viability was detected by MTT method to determine the concentration of D-gal in the follow-up experiment. The cells were divided into the control group, the D-gal group and the Fer-1 group. The cell viability was detected by MTT method. The level of intracellular reactive oxygen species (ROS) was detected by DCFH-DA method. The intracellular glutathione (GSH) content was detected by microplate method. Intracellular malondialdehyde (MDA) content was detected by thiobarbituric acid method. The protein expression levels of SLC7A11, GPx4 and P53 were detected by Western blot assay. The activity of intracellular β-galactosidase (β-GAL) was detected by microassay. Results MTT assay showed that the cell viability decreased with the increase of D-gal concentration (P<0.05). The concentration of D-gal was 20 g/L in subsequent experiments. Compared with the D-gal group, the cell viability was increased in the Fer-1 group (P<0.05). Compared with the control group, ROS and MDA contents, P53 protein expression level and β -GAL activity were increased in the D-gal group, while GSH content, SLC7A11 and GPx4 protein expression levels were decreased (P<0.05). Compared with the D-gal group, ROS and MDA contents, P53 protein expression level and β -GAL activity were decreased in the Fer-1 group, while GSH, SLC7A11 and GPx4 protein expressions were increased (P<0.05). Conclusion Ferroptosis occurs during D-gal-induced cell senescence, and Fer-1 can inhibit iron death and delay myocardial senescence.

Key words: myocytes, cardiac, cell aging, galactose, ferroptosis, D-gal, Ferrostatin-1

CLC Number:

R363.21

Figures/Tables 4

References 29

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组别	ROS	GSH（μg/10⁶ cell）	MDA（nmol/mg prot）
Control组	15.24±1.65	86.01±5.73	0.58±0.05
D-gal组	39.90±4.83^a	63.06±10.98^a	1.46±0.15^a
Fer-1组	26.50±2.66^b	82.59±3.28^b	0.94±0.05^b
F	41.345^＊＊	8.408^＊	67.163^＊＊

组别	ROS	GSH（μg/10⁶ cell）	MDA（nmol/mg prot）
Control组	15.24±1.65	86.01±5.73	0.58±0.05
D-gal组	39.90±4.83^a	63.06±10.98^a	1.46±0.15^a
Fer-1组	26.50±2.66^b	82.59±3.28^b	0.94±0.05^b
F	41.345^＊＊	8.408^＊	67.163^＊＊

组别	SLC7A11	GPx4	P53
Control组	1.04±0.18	1.03±0.18	0.69±0.35
D-gal组	0.48±0.09^a	0.67±0.12^a	1.13±0.05^a
Fer-1组	0.79±0.12^b	0.98±0.07^b	0.88±0.04^b
F	12.706^＊＊	6.730^＊	92.479^＊＊

组别	SLC7A11	GPx4	P53
Control组	1.04±0.18	1.03±0.18	0.69±0.35
D-gal组	0.48±0.09^a	0.67±0.12^a	1.13±0.05^a
Fer-1组	0.79±0.12^b	0.98±0.07^b	0.88±0.04^b
F	12.706^＊＊	6.730^＊	92.479^＊＊