Effects of autonomic nervous regulation on myocardial structural remodeling, electrical remodeling and fibrosis in rats with ejection fraction preserved heart failure based on calcium overload

doi:10.11958/20220897

Abstract

Abstract:

Objective To explore the effects of autonomic nervous regulation on structural remodeling, electrical remodeling and fibrosis in rats with ejection fraction preserved heart failure (HFpEF). Methods A total of 44 SPF grade 12-week-old SD rats were selected, 10 of them were selected as the control group, and the remaining 34 rats were used to establish the HFpEF model. The rat HFpEF model was established by ligation of abdominal aorta-inferior vena cava fistula. Among them, 30 rats were successfully modeled, and 4 rats died. The successfully established rats were divided into 3 groups with 10 rats in each group: the model group, the autonomic nerve regulation group and the autonomic nerve regulation+ acetylcholine M₂ receptor antagonist group. The autonomic nerve regulation group received percutaneous auricular marginal vagus nerve stimulation on the basis treatment of the model group. In the autonomic regulation + acetylcholine M₂ receptor antagonist group, mesotramine (0.5 mg/kg) was injected into the caudal vein every day on the basis treatment of the autonomic nervous regulation group. Left ventricular posterior wall end-diastolic dimension (LVPWD-D), interventricular ventricular septum end-diastolic dimension (IVS-D) and the peak value of mitral flow velocity in the early diastole/peak value of mitral flow velocity in late diastole (E/A) were measured by echocardiography.The effective refractory period (ERP) and monophasic action potential duration (MAPD) of rat hearts were obtained by cardiac electrophysiological stimulator in each group. The value of NT-proBNP was detected by ELISA. The disordered arrangement of cardiomyocytes and the infiltration of inflammatory cells were observed by HE staining. The degree of myocardial fibrosis was observed by Masson staining. The expression levels of matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), sarcoplasmic reticulum Ca²⁺-ATP enzyme (SERCA2a) and its regulatory protein phosphoprotein (PLB) mRNA and protein were detected by RT-PCR and Western blot assay. Results HE staining showed that the arrangement of cardiomyocytes was disordered, the intercellular space was not obvious enough, accompanied by the infiltration of inflammatory cells in the model group. Masson staining showed that the myocardial fibers were disordered, and a large number of collagen fibers were produced in the model group. The above pathological changes were significantly less in the autonomic nervous regulation group than those in the model group. The above pathological changes were not significantly improved in the autonomic nervous regulation + acetylcholine M₂ receptor antagonist group compared with those in the model group. Compared with the control group, the values of NT-proBNP, LVPWD-D, IVS-D, ERP, MAPD, MMP-9 mRNA and protein, PLB mRNA and protein were all increased in the model group, and levels of E/A, TIMP-1 mRNA and protein, SERCA2a mRNA and protein decreased significantly (P<0.05). Compared with the model group, values of NT-proBNP, LVPWD-D, IVS-D, ERP, MAPD, MMP-9 mRNA and protein, PLB mRNA and protein decreased in the autonomic nerve regulation group, and levels of E/A, TIMP-1 mRNA and protein, SERCA2a mRNA and protein were all increased (P<0.05). There were no significant differences in the above-mentioned indexes between the autonomic nervous regulation + acetylcholine M₂receptor antagonist group and the model group. Conclusion Autonomic nervous regulation may reduce structural remodeling, electrical remodeling and myocardial fibrosis in HFpEF model rats and improve the prognosis of HFpEF through reducing the level of calcium overload.

Key words: ventricular remodeling, fibrosis, sarcoplasmic reticulum calcium-transporting ATPases, vagus nerve stimulation, heart failure with preserved ejection fraction, structural remodeling, electrical remodeling, calcium overload

CLC Number:

R541.6

Figures/Tables 8

References 18

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基因名称	引物序列（5′→3′）	产物大小（bp）
MMP-9	上游：AGGTGCCTCGGATGGTTATCG	208
	下游：TGCTTGCCCAGGAAGACGAA	208
TIMP-1	上游：GGCGAACCGGAAACCTGT	188
	下游：GCGCCCTTTGCATCTCTGG	188
SERCA2a	上游：CACCTG?GAA?GAT?TCT?GCG?AAC?T	226
	下游：CAT?CCT?CAT?CCT?GCC?CAA?AG	226
PLB	上游：GGA?GCC?TGT?GTC?CTT?GTG?TC	159
	下游：GAA?GTC?ATC?CCT?GGT?GTC?GT	159
GAPDH	上游：TGCCACTCAGAAGACTGTGG	227
	下游：GGATGCAGGGATGTTCT	227

基因名称	引物序列（5′→3′）	产物大小（bp）
MMP-9	上游：AGGTGCCTCGGATGGTTATCG	208
	下游：TGCTTGCCCAGGAAGACGAA	208
TIMP-1	上游：GGCGAACCGGAAACCTGT	188
	下游：GCGCCCTTTGCATCTCTGG	188
SERCA2a	上游：CACCTG?GAA?GAT?TCT?GCG?AAC?T	226
	下游：CAT?CCT?CAT?CCT?GCC?CAA?AG	226
PLB	上游：GGA?GCC?TGT?GTC?CTT?GTG?TC	159
	下游：GAA?GTC?ATC?CCT?GGT?GTC?GT	159
GAPDH	上游：TGCCACTCAGAAGACTGTGG	227
	下游：GGATGCAGGGATGTTCT	227

组别	NT-proBNP（ng/L）	E/A	LVPWD-D（mm）	IVS-D（mm）	LVEF	ERP（ms）	MAPD（ms）
对照组	45.22±6.26	1.85±0.06	7.59±0.06	1.51±0.02	0.58±0.04	155.43±4.32	172.04±4.85
模型组	301.25±16.07^a	1.28±0.06^a	7.99±0.22^a	2.14±0.08^a	0.57±0.06	183.11±5.83^a	216.95±4.76^a
调控组	79.33±5.63^b	1.66±0.05^b	7.74±0.04^b	1.70±0.02^b	0.57±0.06	167.49±2.23^b	179.86±2.98^b
调控+拮抗组	297.73±18.77	1.28±0.04	7.97±0.20	2.14±0.07	0.58±0.05	182.88±4.90	216.22±4.40
F	1 112.080^＊＊	255.841^＊＊	16.429^＊＊	371.247^＊＊	0.049	87.549^＊＊	301.465^＊＊

组别	NT-proBNP（ng/L）	E/A	LVPWD-D（mm）	IVS-D（mm）	LVEF	ERP（ms）	MAPD（ms）
对照组	45.22±6.26	1.85±0.06	7.59±0.06	1.51±0.02	0.58±0.04	155.43±4.32	172.04±4.85
模型组	301.25±16.07^a	1.28±0.06^a	7.99±0.22^a	2.14±0.08^a	0.57±0.06	183.11±5.83^a	216.95±4.76^a
调控组	79.33±5.63^b	1.66±0.05^b	7.74±0.04^b	1.70±0.02^b	0.57±0.06	167.49±2.23^b	179.86±2.98^b
调控+拮抗组	297.73±18.77	1.28±0.04	7.97±0.20	2.14±0.07	0.58±0.05	182.88±4.90	216.22±4.40
F	1 112.080^＊＊	255.841^＊＊	16.429^＊＊	371.247^＊＊	0.049	87.549^＊＊	301.465^＊＊

组别	MMP-9 mRNA	MMP-9 蛋白	TIMP-1 mRNA	TIMP-1 蛋白
对照组	1.00±0.02	0.22±0.04	1.00±0.02	0.80±0.07
模型组	5.78±0.81^a	0.69±0.06^a	0.13±0.03^a	0.31±0.03^a
调控组	1.37±0.03^b	0.49±0.05^b	0.72±0.16^b	0.71±0.04^b
调控+拮抗组	5.40±1.62	0.70±0.04	0.13±0.02	0.30±0.06
F	79.460^＊＊	220.176^＊＊	296.185^＊＊	269.906^＊＊