Exploring the effect of red ginseng total saponins on cardiomyocyte apoptosis in dilated cardiomyopathy mice based on miR-144-3p/MAPK1 pathway

doi:10.11958/20252283

Abstract

Abstract:

Objective To investigate the effect of red ginseng total saponins (RGTS) on cardiomyocyte apoptosis in dilated cardiomyopathy (DCM) mice by regulating microRNA-144-3p (miR-144-3p)/mitogen activated protein kinase 1 (MAPK1) pathway. Methods The targeting relationship between miR-144-3p and MAPK1 was analyzed via dual luciferase reporter gene assay. Ten C57BL/6J mice were included as the control group, and the transgenic DCM mice were randomly assigned into the DCM group, the captopril group (CAP, 10.1 mg/kg), the low-dose RGTS (L-RGTS) group, the high-dose RGTS (H-RGTS) group (gavage of 6 and 12 g/kg RGTS), the negative control (NC antagomir) group (gavage of 12 g/kg RGTS+tail vein injection of NC antagomir) and the miR-144-3p antagomir group (gavage of 12 g/kg RGTS+tail vein injection of miR-144-3p antagomir), each had 10 mice. The DCM group and the control group were given equal amount of physiological saline by gavage and tail vein injection, once a day for 8 weeks. Echocardiography was used to detect cardiac function in DCM mice. Quantitative real-time fluorescence PCR (qRT-PCR) was used to detect miR-144-3p and MAPK1 mRNA in myocardial tissue. HE staining was used to detect pathological changes in myocardial tissue. TUNEL staining was used to detect cardiomyocyte apoptosis. Western blot assay was used to measure Bax and MAPK1 protein expression in myocardial tissue. Results The control group had a regular arrangement of cardiomyocytes. The arrangement of cardiomyocytes in the DCM group was irregular, with visible cell swelling, necrosis and nuclear condensation. The cardiomyocyte arrangement was relatively regular, and cell swelling was not prominent in the CAP group, the L-RGTS group, the H-RGTS group and the NC antagomir group. The miR-144-3p antagomir group had irregular arrangement of cardiomyocytes. The dual-luciferase reporter gene assay confirmed that miR-144-3p can specifically and negatively regulate MAPK1. Compared with the control group, the left ventricular systolic inner diameter (LVESD), left ventricular diastolic end diameter (LVEDD), expression levels of MAPK1 mRNA and protein, Bax protein and cell apoptosis rate were all increased in the DCM group, while the left ventricular ejection fraction (LVEF), left ventricular short-axis shortening rate (LVFS), and expression level of miR-144-3p were decreased (P<0.05). Compared with the DCM group, the expression levels of LVESD, LVEDD, MAPK1 mRNA and protein, Bax protein, as well as the apoptosis rate were all decreased in the CAP group, the L-RGTS group and the H-RGTS group, while the expression levels of LVEF, LVFS and miR-144-3p were increased (P<0.05). Changes of the above indicators were more significant in the H-RGTS group than those in the L-RGTS group (P<0.05). Compared with the H-RGTS group and the NC antagomir group, the changes of the above indicators were reversed in the miR-144-3p antagomir group (P<0.05). Conclusion RGTS may regulate the miR-144-3p/MAPK1 pathway to inhibit cardiomyocyte apoptosis in DCM mice.

Key words: cardiomyopathy, dilated, mitogen-activated protein kinase 1, apoptosis, red ginseng total saponins, microRNA-144-3p

CLC Number:

R542.2

Figures/Tables 9

References 22

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基因名称	引物序列（5'→3'）	产物大小/bp
miR-144-3p	上游：GGGAGATCAGAAGGTGATT 下游：GTGCAGGGTCCGAGGT	23
MAPK1	上游：ACTGCCAGAGGACGCTGAGAG 下游：ATGTGGTCGTTGCTGAGGTGTTG	187
U6	上游：CGCAAGGATGACACGCAAAT 下游：ATTTGCGTGTCATCCTTGCG	20
GAPDH	上游：GGGAGCCAAAAGGGTCAT 下游：GAGTCCTTCCACGATACCAA	181

基因名称	引物序列（5'→3'）	产物大小/bp
miR-144-3p	上游：GGGAGATCAGAAGGTGATT 下游：GTGCAGGGTCCGAGGT	23
MAPK1	上游：ACTGCCAGAGGACGCTGAGAG 下游：ATGTGGTCGTTGCTGAGGTGTTG	187
U6	上游：CGCAAGGATGACACGCAAAT 下游：ATTTGCGTGTCATCCTTGCG	20
GAPDH	上游：GGGAGCCAAAAGGGTCAT 下游：GAGTCCTTCCACGATACCAA	181

组别	MAPK1-WT	MAPK1-MUT
mimic-NC组	1.00±0.11	0.99±0.32
miR-144-3p mimic组	0.25±0.07	1.06±0.34
t	14.090^＊	0.367

组别	MAPK1-WT	MAPK1-MUT
mimic-NC组	1.00±0.11	0.99±0.32
miR-144-3p mimic组	0.25±0.07	1.06±0.34
t	14.090^＊	0.367

组别	LVESD/mm	LVEDD/mm	LVEF	LVFS
Control组	2.49±0.33	1.67±0.48	0.684±0.070	0.363±0.038
DCM组	3.86±0.48^a	3.28±0.71^a	0.368±0.042^a	0.150±0.018^a
CAP组	2.73±0.49^b	1.82±0.51^b	0.641±0.070^b	0.338±0.037^b
L-RGTS组	3.21±0.52^b	2.65±0.63^b	0.472±0.053^b	0.224±0.030^b
H-RGTS组	2.78±0.34^bc	1.96±0.56^bc	0.625±0.069^bc	0.319±0.036^bc
NC antagomir组	2.83±0.35	1.92±0.55	0.626±0.067	0.318±0.035
miR-144-3p antagomir组	3.69±0.45^de	2.93±0.68^de	0.419±0.050^de	0.180±0.020^de
F	14.650^＊	11.169^＊	41.971^＊	71.712^＊