
==== Front
Heart Vessels
Heart Vessels
Heart and Vessels
0910-8327
1615-2573
Springer Japan Tokyo

38717698
2410
10.1007/s00380-024-02410-9
Original Article
Plasma microRNA-143 and microRNA-145 levels are elevated in patients with left ventricular dysfunction
Murase Hirotaka 1
Minatoguchi Shingo 2
Heishima Kazuki 3
Yasuda Shinji 1
Satake Atsushi 1
Yoshizumi Ryo 1
Komaki Hisaaki 1
Baba Shinya 1
Ojio Shinsuke 1
Tanaka Toshiki 2
Akao Yukihiro 3
Minatoguchi Shinya minatos@gifu-u.ac.jp

1
Okura Hiroyuki 2
1 https://ror.org/0138ysz16 grid.415535.3 Department of Cardiology, Gifu Municipal Hospital, 7-1 Kashimachou, Gifu, 500-8513 Japan
2 https://ror.org/024exxj48 grid.256342.4 0000 0004 0370 4927 Department of Cardiology, Gifu University Graduate School of Medicine, Gifu, Japan
3 https://ror.org/024exxj48 grid.256342.4 0000 0004 0370 4927 United Graduate School of Drug Discovery and Medical Information Science, Gifu University, Gifu, Japan
8 5 2024
8 5 2024
2024
39 10 867876
10 1 2024
25 4 2024
© The Author(s) 2024
2024
https://creativecommons.org/licenses/by/4.0/ Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
MicroRNA(miR)-143 and miR-145 are mainly expressed in vascular smooth muscle cells. However, the relationship between plasma miR-143 or miR-145 levels and the left ventricular (LV) function in patients with heart diseases remains unclear. Blood samples were taken from the antecubital vein in patients with heart diseases (n = 52), such as coronary artery disease, old myocardial infarction, cardiomyopathy, and valvular heart disease, and controls without heart diseases (n = 22). We measured plasma miR-143 and -145 levels by quantitative RT–PCR using TaqMan MicroRNA Assays and THUNDERBIRD Probe qPCR Mix. Plasma BNP levels were also measured. Echocardiography was performed to measure the LV ejection fraction (LVEF) and LV dilation. Plasma miR-143 and miR-145 levels were significantly higher in patients with heart diseases than in controls, respectively. Plasma miR-143 and miR-145 levels were significantly higher in patients with LVEF < 50% than in those with LVEF ≧ 50%, respectively. Plasma miR-143 and miR-145 levels were inversely correlated with LVEF, respectively. Plasma miR-143 and miR-145 levels were positively correlated with LV end-systolic dimension, respectively. Plasma miR-143 and -145 levels were positively correlated with plasma BNP levels, respectively. Plasma BNP levels were inversely correlated with LVEF. Plasma miR-143 and miR-145 levels are elevated in patients with LV dysfunction and may counteract LV dysfunction.

Keywords

Microrna-143
microRNA-145
LV dysfunction
LV dilation
Heart diseases
Gifu UniversityOpen Access funding provided by Gifu University.

issue-copyright-statement© Founding Editors’ Group 2024
==== Body
pmcIntroduction

In accordance with the increased number of elderly people, the number of patients with heart failure is markedly increasing in Japan [1]. Heart failure is the end-stage phenotype of several heart diseases, such as coronary artery disease, old myocardial infarction, valvular heart disease, congenital heart disease, cardiomyopathy, and hypertension. The plasma brain natriuretic peptide (BNP) level is increased in patients with heart failure and is a powerful predictor of the prognosis associated with symptomatic and asymptomatic heart failure [2, 3].

The microRNAs (miRs) are 21–25 base non-coding RNAs, and approximately 2700 miRs have been identified in humans (miRBase, http://www.mirbase.org/). The miRs negatively regulate messenger RNA (mRNA) expression by inhibiting translation or degrading mRNAs [4]. It has been reported that miRs play an important role in a variety of biological processes, such as cell death, cell proliferation, and cell differentiation [5]. Recently, some of miRs have been reported to serve as a potential biomarker for heart failure [6, 7]. Among many miRs, miR-143 and miR-145 are associated with the proliferation of vascular smooth muscle cells [8] and progression of atherosclerosis [9]. It was reported that miR-143 and miR-145 are expressed in the heart [8, 10, 11], miR-143 regulates morphogenesis of the heart [11], and miR-145 regulates hypertrophy of the heart [11]. Therefore, miR-143 and miR-145 may regulate the pathophysiology of heart diseases [12]. We previously reported that the intravenous administration of miR-145 after acute myocardial infarction (AMI) reduces the myocardial infarct size and improves the left ventricular (LV) function as compared with controls in rabbits [13], and we recently reported that plasma miR-143 and miR-145 levels increase in the acute phase of AMI and that the increase in plasma miR-143 levels positively correlated with recovery of LV function and the increase in plasma miR-145 tended to positively correlate with recovery of LV function in the chronic phase at 6 months in patients with AMI [14].

Since miR-143 and miR-145 have been reported to be located approximately 1.3 kb from each other on chromosome 5q33 and have similar characteristics [15], both miR-143 and miR-145 may contribute to repair damaged cardiac tissue and improve the deteriorated LV function in patients with heart diseases. However, the behavior of plasma miR-143 and miR-145 levels and association between plasma miR-143 or miR-145 levels and LV function in the chronic state have not yet to be clarified in patients with heart diseases. Thus, in the present study, we aimed to investigate the relationship between plasma miR-143 or miR-145 levels and the LV function, between plasma miR-143 or miR-145 levels and LV dilation, and between plasma miR-143 or miR-145 levels and the plasma BNP level, a well-known marker of heart failure, in patients with heart diseases.

Subjects and methods

The protocol of the present study was approved by the ethics committee of Gifu University Graduate School of Medicine (Approval number: 30-011) and Gifu Municipal Hospital (Approval number: 455). The investigation conformed with the principles outlined in the Declaration of Helsinki (Br Med J 1964; ii:177). The public and trial registry number is UMIN000040165.

Study patients

This study included a total of 74 patients who underwent cardiac catheterization and echocardiography at Gifu City Hospital and Gifu University Hospital to investigate cardiac disease in patients with precordial complaints and breathing difficulties. Control group (n = 22) consists of patients without significant coronary artery stenosis or left ventricular damage, and some of them were previously performed percutaneous coronary artery intervention and receiving some drugs. In contrast, heart disease group (n = 52) consists of patients with significant coronary stenosis, old myocardial infarction, cardiomyopathy and valvular heart disease. The subjects consisted of 30 males and 44 females, with a mean age of 71.4 ± 10.7 years. The study period was from July 2018 to June 2021.

Echocardiography

The LV ejection fraction (LVEF), LV end-diastolic dimension (LVDd), and LV end-systolic dimension (LVSd) were obtained by echocardiography (iE33, PHILIPS, Tokyo, Japan). We used the modified Simpson’s method, which is regarded as a reliable method to estimate LVEF. An echocardiologist performed the echocardiography, who was blinded to the protocol of this study. Echocardiography was performed before the day of cardiac catheterization.

Measurements of plasma miR-143 and miR-145 levels, and plasma BNP levels

The timing of blood sampling for miR assay was on the day of cardiac catheterization. Blood samples were taken from the antecubital vein and collected into sterile tubes containing EDTA, immediately placed on ice, and then centrifuged at 1500 g for 15 min. Plasma was then collected and frozen at  – 83 °C until further analysis. To minimize RNA degradation, we only used samples that were freeze‐thawed once. MiRNAs from frozen plasma were extracted using the NucleoSpin miRNA Plasma kit (MACHEREY‐NAGEL GmbH, Düren, Germany) according to the standard protocol. Proteins in the supernatant were precipitated using a reagent in the kit and removed by centrifugation. After adjustment of the binding conditions with isopropanol, miRNAs were bound to a miRNA collection column. The miRNA quality was assessed by measuring circulating miR16‐5p levels. The purified miRNA was immediately used for reverse transcription to prevent degradation of RNA before the PCR step. To determine plasma miR-143 and miR-145 levels, we conducted quantitative RT–PCR (qRT–PCR) using TaqMan microRNA assays and THUNDERBIRD Probe qPCR Mix.

Since miR16‐5p is abundantly and constantly found in plasma of the control and heart failure patients, similar to ribosomal RNA in cellular RNAs, we used miR16-5p as an internal control. Therefore, its levels reflect miRNA degradation and the quality of plasma samples. As to this, we cited 5 papers from our laboratory and other laboratories [14, 16–19]. Plasma miR-143 and miR-145 levels are expressed relative to miR-16 as an internal control and expressed as ∆Ct. The repeatability for measurement of miRNA levels was confirmed by assessing the same samples multiple times (average coefficient of variation less than 0.25, n = 4) [14]. Plasma BNP levels were measured by the Shionoria BNP RIA kit (Shionogi, Osaka, Japan).

Blood biochemical analysis

Blood samples were taken from the antecubital veins. Creatinine, total-cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglycerides (TG), and hemoglobin A1c (HbA1C) were measured.

Complications and drugs used

Complications such as hypertension, diabetes mellitus, hyperlipidemia, and drugs used were examined.

Statistical analysis

Data are shown as the mean ± standard deviation (SD). Categorical data are summarized as percentages and compared with a Chi-square test or Fisher’s exact test, as appropriate. The normality of data distributions was tested using the Kolmogorov–Smirnov test. The significance of differences between groups for variables that were normally distributed was determined by the unpaired Student’s t test. Correlation coefficients between two variables were obtained by linear regression analysis using Pearson’s correlation analysis. These statistical analyses were performed using GraphPad Prism 7 (GraphPad Software Inc.). A p value < 0.05 was considered significant, and p < 0.01 and p < 0.001 were considered highly significant. Factors for LVEF and LVSd were analyzed by using logistic regression models. Multivariate logistic regression analysis (forward selection, based on the likelihood ratio, or forced entry method) was performed. A two-sided p value of less than 0.05 was considered statistically significant. Statistical analysis was made with SPSS for Windows version 22.0 (IBM Japan, Tokyo, Japan).

Results

Patients’ characteristics and drugs used

Patients’ characteristics are shown in Table 1. In the control group (n = 22, male: n = 4, female: n = 18), the mean age was 73.9 ± 7.5 years. In the heart disease group (n = 52, male: n = 26 female: n = 26), the mean age was 70.6 ± 11.5 years. Heart diseases consisted of coronary artery disease (n = 19), old myocardial infarction (n = 23), valvular heart disease (n = 5), cardiomyopathy (n = 2), and arrhythmias (n = 3), and controls were subjects without heart diseases (n = 22). Biochemical data are shown in Table 1. Drugs used were ACE I/ARBs (n = 39), CCBs (n = 33), beta-blockers (n = 30), statins (n = 41), insulin (n = 4), DPP4 inhibitors (n = 15), SGLT2 inhibitors (n = 5), Metoformin (n = 11), antiplatelets (n = 42), and DOAC (n = 9). Complications were hypertension (n = 49), dyslipidemia (n = 35), and diabetes mellitus (n = 32).Table 1 Patient’s characteristics and drugs used

	Control group (n = 22)	Cardiac disease group (n = 52)	P value	
Age, years	73.9 ± 7.5	70.6 ± 11.5	0.1552	
Sex, (n)	M/F, 4/18	M/F, 26/26	0.0185	
Subjects, n				
HTN, n(%)	17 (77.3)	32 (61.5)	0.2827	
HL, n(%)	11 (50.0)	24 (46.2)	0.8032	
DM, n(%)	8 (36.4)	24 (46.2)	0.6083	
Biochemical data				
Creatinine (mg/dL)	1.03 ± 0.96	0.92 ± 0.36	0.6133	
TC (mg/dL)	181 ± 26.4	181 ± 36.6	0.9898	
LDL-C (mg/dL)	102 ± 19.1	104 ± 32.4	0.83	
HDL-C (mg/dL)	48.9 ± 14.3	53.9 ± 16.5	0.221	
TG (mg/dL)	138 ± 62.0	156 ± 109	0.3929	
HbA1c (%)	6.2 ± 0.71	6.4 ± 0.91	0.6601	
Drug used, n(%)				
ACEI/ARB	9 (40.9)	30 (57.7)	0.2116	
CCB	15 (68.2)	18 (34.6)	0.0107	
Beta blocker	7 (31.8)	23 (44.2)	0.4383	
Statin	12 (54.5)	26 (50.0)	0.8019	
Insulin	1 (4.5)	3 (5.8)	0.831	
DPP4-Inhibitor	3 (13.6)	12 (23.1)	0.5293	
SGLT2-Inhibitor	0 (0)	5 (9.6)	0.3133	
Metformin	4 (18.2)	7 (13.5)	0.7227	
Antiplatelet	12 (54.5)	30 (57.7)	0.8038	
DOAC	2 (9.1)	7 (13.5)	0.599	
HTN  hypertension, HL  hyperlipidemia, DM  diabetes mellitus, TC  total cholesterol, LDL-C  low density lipoprotein cholesterol, HDL-C  high density lipoprotein cholesterol, TG  triglyceride, CCB  calcium channel blocker, DOAC  direct oral anticoagulant

Plasma miR-143 and miR-145 levels in controls and heart disease patients

Plasma miR-143 levels were significantly higher in the heart disease group than in the control group (p = 0.0423) (Fig. 1A). Plasma miR-145 levels were also significantly higher in the heart disease group than in the control group (p = 0.0063) (Fig. 1B).Fig. 1 A Plasma miR-143 levels in control and heart disease groups. B Plasma miR-145 levels in control and heart disease groups. C Relationship between plasma miR-143 levels and plasma miR-145 levels

Relationship between plasma miR-143 levels and plasma miR-145 levels

Plasma miR-143 levels were closely and positively correlated with plasma miR-145 levels (Fig. 1C).

Plasma miR-143 and miR-145 levels between LVEF≧50% and LVEF < 50%

Plasma miR-143 levels were significantly higher in the LVEF < 50% group than in the LVEF≧50% group (p = 0.0062) (Fig. 2A). Plasma miR-145 levels were also significantly higher in the LVEF < 50% group than in the LVEF≧50% group (p = 0.0339) (Fig. 2B).Fig. 2 A Plasma miR-143 levels in LVEF≧50% and < 50% groups. B Plasma miR-145 levels in LVEF≧50% and < 50% groups

Relationship between plasma miR-143 or miR-145 levels and LVEF

Plasma miR-143 levels were inversely correlated with LVEF (n = 74, r = 0.3874, p = 0.0006) (Fig. 3A). Plasma miR-145 levels were also inversely correlated with LVEF (n = 74, r = 0.3512, p = 0.0022) (Fig. 3B).Fig. 3 A Relationship between plasma miR-143 levels and LVEF. B Relationship between plasma miR-145 levels and LVEF

Relationship between plasma miR-143 or miR-145 levels and LV dilation

Plasma miR-143 levels were positively correlated with the LV end-systolic dimension (LVSd), an indicator of LV remodeling (n = 74, r = 0.302, p = 0.0089) (Fig. 4A). The plasma miR-145 levels were also positively correlated with LVSd (n = 74, r = 0.242, p = 0.0377) (Fig. 4B). Plasma miR-143 levels were not correlated with the LV end-diastolic dimension (LVDd) (n = 74, r = 0.1751, p = 0.1356) (Fig. 4C). Plasma miR-145 levels were not correlated with LVDd (n = 74, r = 0.1339, p = 0.2553) (Fig. 4D).Fig. 4 The A Relationship between plasma miR-143 levels and LVSd. B Relationship between plasma miR-145 levels and LVSd. C Relationship between plasma miR-143 levels and LVDd. D Relationship between plasma miR-145 levels and LVDd

Relationship between plasma BNP levels and LVEF, plasma miR-143 levels, or plasma miR-145 levels

Plasma BNP levels were inversely correlated with LVEF (n = 69, r = 0.3366, p = 0.0047) (Fig. 5A). Plasma miR-143 levels were positively correlated with plasma BNP levels (n = 69, r = 0.3748, p = 0.0015) (Fig. 5B). Plasma miR-145 levels were also positively correlated with plasma BNP levels (n = 69, r = 0.4726, p < 0.0001) (Fig. 5C).Fig. 5 The A Relationship between plasma BNP levels and LVEF. B: Relationship between plasma miR-143 and plasma BNP levels. C: Relationship between plasma miR-145 and plasma BNP levels

Factors that may affect LVEF and LVSd

Many of the factors that may affect LVEF and LVSd were compared between the LVEF ≧50% and LVEF < 50% groups and between LVSd > 42 mm and LVSd≦42 mm groups (Tables 2, 3). Among many factors, univariate analysis showed a significant difference in miR-143 (p = 0.0062) and miR-145 (p = 0.0339) for LVEF (Table 2) and showed a significant difference in miR-143 (p = 0.0034) and miR-145 (p = 0.0469) for LVSd (Table 3). Multivariate logistic regression analysis showed a significant difference (OR = 1.178, 95% CI 1.025–1.355, p = 0.021) in miR143 for LVEF (Table 2), and showed a significant difference (OR = 1.196, 95% CI 1.030–1.389, p = 0.019) in miR-143 for LVSd (Table 3). Therefore, among many factors, only miR-143 was more strongly correlated with LVEF and LVSd than miR-145.Table 2 Comparison of factors that affect LVEF

	LVEF≧50%
(n = 57)	LVEF < 50%
(n = 17)	P value	
miR-143 (pg/ml)	0.0526 ± 0.0273	0.0835 ± 0.0667	0.0062	
miR-145 (pg/ml)	0.0428 ± 0.0317	0.0650 ± 0.0521	0.0339	
Characteristics				
Age (years)	73 ± 10.6	74 ± 11.2	0.7152	
Sex, (n)	M/F, 23/34	M/F, 7/10	1	
HTN, n(%)	41 (71.9)	8 (47.1)	0.0801	
HL, n(%)	28 (49.1)	7 (47.1)	0.593	
DM, n(%)	22 (38.6)	10 (58.8)	0.1695	
Biochemical data				
Creatinine (mg/dL)	0.95 ± 0.67	0.97 ± 0.32	0.893	
TC (mg/dL)	181 ± 33.7	182 ± 33.5	0.9689	
LDL-C (mg/dL)	105 ± 29.2	100 ± 32.3	0.6732	
HDL-C (mg/dL)	53.9 ± 17.2	46.2 ± 9.4	0.1145	
TG (mg/dL)	144 ± 70.0	170 ± 159	0.3567	
HbA1c (%)	6.25 ± 0.84	6.57 ± 0.92	0.2153	
Medications used, n(%)				
ACEI/ARB	30 (52.6)	9 (52.9)	1	
CCB	31 (54.4)	2 (11.8)	0.002	
Beta blocker	21 (36.8)	9 (52.9)	0.2696	
Statin	30 (52.6)	8 (47.1)	0.785	
Insline	3 (5.3)	1 (5.9)	1	
DPP4-Inhibitor	10 (17.5)	5 (29.4)	0.313	
SGLT2-Inhibitor	2 (3.5)	3 (17.6)	0.0762	
Metphrmine	10 (17.5)	1 (5.9)	0.4385	
Antiplatelet	33 (57.9)	9 (52.9)	0.7842	
DOAC	5 (8.8)	4 (23.5)	0.1972	
HTN  hypertension, HL  hyperlipidemia, DM  diabetes mellitus, TC  total cholesterol, LDL-C  low density lipoprotein cholesterol, HDL-C  high density lipoprotein cholesterol, TG  triglyceride, CCB calcium channel blocker, DOAC  direct oral anticoagulant

Table 3 Comparison of factors that affect LVSD

	LVSd≦42 mm
(n = 68)	LVSd > 42 mm
(n = 6)	P value	
miR-143 (pg/ml)	0.0556 ± 0.0453	0.1063 ± 0.0847	0.0034	
miR-145 (pg/ml)	0.0453 ± 0.0351	0.0774 ± 0.0595	0.0469	
Characteristics				
Age (years)	73 ± 10.4	71 ± 13.7	0.422	
Sex, (n)	M/F, 26/42	M/F, 4/2	0.2151	
HTN, n(%)	47 (69.1)	2 (33.3)	0.1708	
HL, n(%)	32 (47.1)	3 (50.0)	1	
DM, n(%)	29 (42.6)	3 (50.0)	1	
Biochemical data				
Creatinine (mg/dL)	0.94 ± 0.62	1.13 ± 0.38	0.469	
TC (mg/dL)	180 ± 33.2	194 ± 36.4	0.379	
LDL-C (mg/dL)	103 ± 29.7	110 ± 32.9	0.692	
HDL-C (mg/dL)	52.2 ± 16.6	53.0 ± 7.97	0.917	
TG (mg/dL)	148 ± 99.3	169 ± 81.6	0.633	
HbA1c (%)	6.31 ± 0.85	6.40 ± 1.11	0.817	
Medications used, n(%)				
ACEI/ARB	35 (51.5)	4 (66.7)	0.677	
CCB	33 (48.5)	0 (0)	0.0303	
Beta blocker	27 (39.7)	3 (50.0)	0.6812	
Statin	35 (51.5)	3 (50.0)	1	
Insline	4 (5.9)	0 (0)	1	
DPP4-Inhibitor	13 (19.1)	2 (33.3)	0.5946	
SGLT2-Inhibitor	4 (5.9)	1 (16.7)	0.3529	
Metphrmine	11 (16.2)	0 (0)	0.5826	
Antiplatelet	40 (58.8)	2 (33.3)	0.3929	
DOAC	8 (11.8)	1 (16.7)	0.5541	
HTN  hypertension, HL  hyperlipidemia, DM  diabetes mellitus, TC  total cholesterol, LDL-C  low density lipoprotein cholesterol, HDL-C  high density lipoprotein cholesterol, TG  triglyceride, CCB  calcium channel blocker, DOAC  direct oral anticoagulant

Discussion

The findings of the present study were that: (1) plasma miR-143 and miR-145 levels were significantly higher in patients with heart diseases than controls, respectively, (2) plasma miR-143 and miR-145 levels were significantly higher in patients with LVEF < 50% than in those with LVEF ≧ 50%, respectively, (3) plasma miR-143 and miR-145 levels were inversely correlated with LVEF, respectively, (4) plasma miR-143 and miR-145 levels were positively correlated with the LV end-systolic dimension (LVSd), respectively, (5) plasma BNP levels were positively correlated with LVEF, and (6) plasma miR-143 and miR-145 levels were positively correlated with plasma BNP levels, respectively.

It has been reported that miR-143 and miR-145 are involved in the pathogenesis of many cardiovascular diseases, such as essential hypertension [20, 21], atherosclerosis [9, 22, 23], coronary artery disease [24, 25], and myocardial infarction [13]. MiR-143 and miR-145 are highly expressed in vascular smooth muscle cells [8, 23, 26] and the heart [8, 10]. MiR-143 and miR-145 are present in the peripheral blood in the form of a nuclease-resistant complex with argonaute (AGO) protein [27] or in exosomes [28] released from the cells containing miR-143 and miR-145, such as vascular smooth muscle cells and cardiomyocytes. However, the behavior of plasma miR-143 and -145 levels in cardiac patients with LV dysfunction has not still been fully clarified. Therefore, the measurement of plasma levels of miR-143 and miR-145 may help to understand the roles of miR-143 and miR-145 in the pathophysiology of LV dysfunction in patients with heart diseases. As a matter of fact, other studies reported that miR-143 and miR-145 were associated with cardiovascular diseases [29] and that plasma miR-143 and miR-145 levels increased in patients with advanced heart failure [30].

In the present study, plasma miR-143 and miR-145 levels were significantly higher in the heart disease group than in the control group, respectively (Fig. 1A, B). Plasma miR-143 and miR-145 levels were significantly higher in patients with LVEF < 50% than in those with LVEF≧50%, respectively (Fig. 2A, B), suggesting that the LV function is a determinant factor of plasma miR-143 and miR-145 levels; patients with LV dysfunction show higher plasma miR-143 and miR-145 levels, and those with a normal LV function show lower plasma miR-143 and miR-145 levels. Furthermore, we investigated the relationship between plasma miR-143 levels or plasma miR-145 levels and LVEF. As a result, plasma miR-143 and miR-145 levels were inversely correlated with LVEF, respectively (Fig. 3A, B). These results suggest that plasma miR-143 and miR-145 levels increase depending on the severity of deterioration of LV function; lower plasma miR-143 and miR-145 levels were associated with a normal LV function, and higher plasma miR-143 and miR-145 levels were associated with a deteriorated LV function.

Regarding LV chamber dilation, plasma miR-143 and miR-145 levels were positively correlated with the LV end-systolic dimension (LVSd) (p = 0.0089 and p = 0.0037, respectively) (Fig. 4A, B). However, there was no correlation between plasma 143 levels or plasma miR-145 levels and the LV end-diastolic dimension (LVDd) (Fig. 4C, D). These results suggest that plasma miR-143 and miR-145 levels increase depending on the dilation of LVSd: lower plasma miR-143 and miR-145 levels were associated with smaller LVSd, and higher plasma miR-143 and miR-145 levels were associated with greater LVSd. Since LVSd has been reported to be a good indicator of LV remodeling [31], plasma miR-143 and miR-145 levels may be associated with LV remodeling. We previously reported that the intravenous administration of miR-145 reduced the infarct size, improved the cardiac function, and attenuated LV remodeling in a rabbit model of acute myocardial infarction [13]. Since it has been reported that miR-143 and miR-145 are located approximately 1.3 kb from each other on chromosome 5q33 and have similar characteristics [15], both miR-143 and miR-145 might have similar abilities to repair damaged cardiac tissue, improve the deteriorated LV function, and attenuate LV remodeling in patients with LV dysfunction. According to the results of the animal experiment and assumption that miR-143 and miR-145 have similar effects on the heart, LV dysfunction and LV remodeling by themselves might have facilitated the release of endogenous miR-143 and miR-145 into the peripheral blood from cells in cardiac and vascular tissues, leading to the increase in plasma miR-143 and miR-145 levels.

The plasma BNP level is regarded as a diagnostic and prognostic marker of symptomatic and asymptomatic heart failure [2, 3]. In the present study, plasma BNP levels were inversely correlated with LVEF, an indicator of the LV function (Fig. 5A), suggesting that higher plasma BNP levels are associated with lower LVEF, and lower plasma BNP levels are associated with higher LVEF.

In the present study, the behaviors of plasma BNP, miR-143, and miR-145 levels were similar in terms of the relationship with LVEF. Plasma BNP, miR-143, and miR-145 levels were inversely correlated with LVEF (Figs 3A, B, 5A). In addition, there was a positive correlation between plasma BNP and miR-143 levels (p = 0.0015) (Fig. 5B), and between plasma BNP and miR-145 levels (p < 0.0001) (Fig. 5C). These results suggest that plasma miR-143 and miR-145 levels may be also diagnostic and prognostic markers of heart failure. BNP by itself has been reported to improve the cardiac function in a rat model of heart failure [32]. Angiotensin receptor–neprilysin inhibitor (ARNI), a combination of sacubitril and valsartan, is a new drug for heart failure. The PARADIGM–HF trial, in which 8,442 heart failure patients were enrolled in a double-blind and randomized trial, demonstrated that ARNI was superior to enalapril in reducing the risks of death and hospitalization for heart failure [33]. In the PARADIGM–HF trial, the blockade of neprilysin by sacubitril reduced the degradation of natriuretic peptides, resulting in an increase of plasma BNP levels [34]. Based on these studies, elevated plasma BNP levels in heart failure patients are considered to counteract LV dysfunction.

Because the present study is a clinical study demonstrating the relationship between plasma miR-143 or miR-145 levels and LV dysfunction or LV dilation, it is difficult to show the precise mechanistic insight how miR-143 and miR-145 are upregulated in heart diseases in the clinical setting. However, it has been reported that miR-143 and miR-145 can be upregulated in endothelial cells in response to shear stress and subsequently exported in exosome-like vesicles that regulate vascular smooth muscle phenotype [35]. Furthermore, in the present study, plasma miR-143 or miR-145 levels positively correlated with plasma BNP levels (Fig. 5B, C). Since BNP is regarded as a marker of intravascular volume, and BNP secretion has a direct linear correlation with intravascular volume status [36], higher plasma BNP levels mean higher intravascular volume status, leading to a greater shear stress to the vascular endothelial cells, and then upregulation of miR-143 and miR-145. Furthermore, since plasma miR-143 levels were closely and positively correlated with plasma miR-145 levels in the present study (Fig. 1C), plasma miR-143 and miR-145 levels might have behaved similarly in heart diseases due to the shear stress to the vascular endothelial cells. Therefore, one of the mechanisms by which both plasma miR-143 and miR-145 levels are elevated in patients with LV dysfunction may be related to the higher plasma BNP levels in patients with LV dysfunction.

It was previously reported that plasma miR-143 levels were increased in patients with heart failure via induction by HIF-1 [37], a direct stimulator for BNP induction [38]. This report may explain the result in the present study that higher plasma BNP levels were positively correlated with higher plasma miR-143 levels in patients with heart diseases. In the present study, however, both plasma miR-143 and miR-145 levels were significantly higher in patients with heart diseases than those in the controls. Since plasma miR-143 levels positively and closely correlated with plasma niR-145 levels (Fig. 1C), it is reasonable to consider that plasma miR-143 and miR-145 levels behave similarly in patients with heart diseases as shown in the present study.

Furthermore, we recently reported in a clinical study that plasma miR-143 and miR-145 levels increased in the acute phase (within 1 week) of acute myocardial infarction and the increase in plasma miR-143 levels in the acute phase were positively correlated with the increase in LVEF in the chronic phase of 6 months, and the increase in plasma miR-145 levels in the acute phase tended to be positively correlated with the increase in LVEF in the chronic phase of 6 months in patients with acute myocardial infarction [14]. These results mentioned above suggest that higher plasma miR-143 and miR-145 levels may improve LV function and may counteract LV dysfunction in patients with heart diseases. Furthermore, among many factors, univariate analysis demonstrated that miR-143 and miR-145 were correlated with LVEF and LVSd (Tables 2, 3).

However, multivariate logistic regression analysis demonstrated that only miR-143 was correlated with LVEF and LVSd, suggesting that miR-143 was more strongly correlated with LVEF and LVSd than miR-145.

In the present study, the target molecules for miR-143 and miR-145 in patients with heart diseases have not been clarified. However, in an acute myocardial infarction model, it has been reported that the mechanisms by which miR-145 improves the cardiac function involving acceleration of autophagy of cardiomyocytes through targeting fibroblast growth factor receptor substrate 2 (FRS2) [13], and that the mechanism by which miR-143 improves cardiac function is through decreasing oxidative stress that causes autophagy cell death by silencing COX-1, COX-2 and ATG7 [14]. The other study demonstrated the CHK2/Beclin2 pathway as a target molecule to avoid autophagic cell death induced by ischemic heart disease [39].

In conclusion, plasma miR-143 and miR-145 levels increase in heart diseases patients with LV dysfunction. Plasma miR-145 and -143 may counteract LV dysfunction.

Acknowledgements

We thank Mrs Kaori Osawa and Miss Akiko Tsujimoto for technical assistance. Shinya Minatoguchi is a member of Circulation Reports’ of Editorial Team.

Author contributions

Shinya M designed the experiment, HM, Shingo M, SY, AS, RY, HK, SB, TT, SO collected the data, KH and YA measured plasma miRNA levels and HO performed data interpretation, and HM and Shinya M wrote the manuscript.

Funding

Open Access funding provided by Gifu University. This study was supported by funding from Gifu University Graduate School of Medicine (to Shinya Minatoguchi).

Data availability

The deidentified participant data will not be shared.

Declarations

Conflict of interest

The authors declare that they have no conflict of interest.

IRB information

This study was approved by the Ethics Committee of Gifu University Graduate School of Medicine (Approval number: 30-011) and Gifu Municipal Hospital (Approval number: 455).

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
==== Refs
References

1. Shimokawa H Miura M Nochioka K Sakata Y Heart failure as a general pandemic in Asia Eur J Heart Fail 2015 17 884 892 10.1002/ejhf.319 26222508
Shimokawa H, Miura M, Nochioka K, Sakata Y (2015) Heart failure as a general pandemic in Asia. Eur J Heart Fail 17:884–89226222508 10.1002/ejhf.319
2. Tsutamoto T Wada A Maeda K Effects of spilonolactone on plasma brain natriuretic peptide and left ventricular remodeling in patients with congestive heart failure J Am Coll Cardiol 2001 37 1228 1233 10.1016/S0735-1097(01)01116-0 11300427
Tsutamoto T, Wada A, Maeda K (2001) Effects of spilonolactone on plasma brain natriuretic peptide and left ventricular remodeling in patients with congestive heart failure. J Am Coll Cardiol 37:1228–123311300427 10.1016/S0735-1097(01)01116-0
3. Tsutamoto T Wada A Maeda K Hisanaga T Mabuchi N Hayashi M Ohnishi M Sawaki M Fujii M Horie H Sugimoto Y Kinoshita M Plasma brain natriuretic peptide level as a biochemical marker of morbidity and mortality in patients with asymptomatic or minimally symptomatic left ventricular dysfunction Eur Heart J 1999 20 1799 1807 10.1053/euhj.1999.1746 10581138
Tsutamoto T, Wada A, Maeda K, Hisanaga T, Mabuchi N, Hayashi M, Ohnishi M, Sawaki M, Fujii M, Horie H, Sugimoto Y, Kinoshita M (1999) Plasma brain natriuretic peptide level as a biochemical marker of morbidity and mortality in patients with asymptomatic or minimally symptomatic left ventricular dysfunction. Eur Heart J 20:1799–180710581138 10.1053/euhj.1999.1746
4. Bartel DP MicroRNAs: genomics, biogenesis, mechanism, and function Cell 2004 116 281 297 10.1016/S0092-8674(04)00045-5 14744438
Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–29714744438 10.1016/S0092-8674(04)00045-5
5. Ambros V The functions of animal microRNAs Nature 2004 431 350 355 10.1038/nature02871 15372042
Ambros V (2004) The functions of animal microRNAs. Nature 431:350–35515372042 10.1038/nature02871
6. Zou Y Yin J Zhang Q MIR-568 mitigated cardiomyocytes apoptosis, oxidative stress response and cardiac dysfunction via targeting SMURF2 in heart failure rats Heart Vessels 2023 38 857 868 10.1007/s00380-022-02231-8 36717388
Zou Y, Yin J, Zhang Q (2023) MIR-568 mitigated cardiomyocytes apoptosis, oxidative stress response and cardiac dysfunction via targeting SMURF2 in heart failure rats. Heart Vessels 38:857–86836717388 10.1007/s00380-022-02231-8
7. Yang W Han Y Yang C Chen Y Zhao W Su X Yang K Jin W MicroRNA-19b-1 reverses ischaemia-induced heart failure by inhibiting cardiomyocyte apoptosis and targeting Bcl2 I11/BIM Heart Vessels 2019 34 1221 1229 10.1007/s00380-018-01336-3 30607541
Yang W, Han Y, Yang C, Chen Y, Zhao W, Su X, Yang K, Jin W (2019) MicroRNA-19b-1 reverses ischaemia-induced heart failure by inhibiting cardiomyocyte apoptosis and targeting Bcl2 I11/BIM. Heart Vessels 34:1221–122930607541 10.1007/s00380-018-01336-3
8. Cordes KR Sheehy NT White M Berry E Morton SU Muth AN Lee TH Miano JM Ivey KN Srivastava D miR-145 and miR-143 regulate smooth muscle cell fate decisions Nature 2009 460 705 710 10.1038/nature08195 19578358
Cordes KR, Sheehy NT, White M, Berry E, Morton SU, Muth AN, Lee TH, Miano JM, Ivey KN, Srivastava D (2009) miR-145 and miR-143 regulate smooth muscle cell fate decisions. Nature 460:705–71019578358 10.1038/nature08195
9. Sala F Aranda JF Rotllan N Ramírez CM Aryal B Elia L Condrelli G Catapano AL Fernandez-Hernando C Norata GD MiR-143/145 deficiency attenuates the progression of atherosclerosis in Ldlr-/-mice Thromb Haemost 2014 112 796 802 10.1160/TH13-11-0905 25008143
Sala F, Aranda JF, Rotllan N, Ramírez CM, Aryal B, Elia L, Condrelli G, Catapano AL, Fernandez-Hernando C, Norata GD (2014) MiR-143/145 deficiency attenuates the progression of atherosclerosis in Ldlr-/-mice. Thromb Haemost 112:796–80225008143 10.1160/TH13-11-0905
10. Deacon DC Nevis KR Cashman TJ Xhou Y Zhao L Washko D Guner-Ataman B Burns CG Burns CE The miR-143-adducin3 pathway is essential for cardiac chamber morphogenesis Development 2010 137 1887 1896 10.1242/dev.050526 20460367
Deacon DC, Nevis KR, Cashman TJ, Xhou Y, Zhao L, Washko D, Guner-Ataman B, Burns CG, Burns CE (2010) The miR-143-adducin3 pathway is essential for cardiac chamber morphogenesis. Development 137:1887–189620460367 10.1242/dev.050526
11. Li R Yan G Zhang Q Jiang Y Sun H Hu Y Sun J Xu B miR-145 inhibits isoproterenol-induced cardiomyocyte hypertrophy by targeting the expression and localization of GATA 6 FEBS Lett 2013 587 1754 1761 10.1016/j.febslet.2013.04.018 23624080
Li R, Yan G, Zhang Q, Jiang Y, Sun H, Hu Y, Sun J, Xu B (2013) miR-145 inhibits isoproterenol-induced cardiomyocyte hypertrophy by targeting the expression and localization of GATA 6. FEBS Lett 587:1754–176123624080 10.1016/j.febslet.2013.04.018
12. Zhao W Zhao SP Zhao YH MicroRNA-143/-145 in cardiovascular diseases Biomed Res Int 2015 2015 531740 10.1155/2015/531740 26221598
Zhao W, Zhao SP, Zhao YH (2015) MicroRNA-143/-145 in cardiovascular diseases. Biomed Res Int 2015:53174026221598 10.1155/2015/531740
13. Higashi K Yamada Y Minatoguchi S Baba S Iwasa M Kanamori H Kawasaki M Nishigaki K Takemura G Kumazaki M Akao Y Minatoguchi S MicroRNA-145 repairs infarcted myocardium by accelerating cardiomyocyte. Autophagy Am J Physiol Heart Circ Physiol 2015 309 H1813 H1826 10.1152/ajpheart.00709.2014 26432843
Higashi K, Yamada Y, Minatoguchi S, Baba S, Iwasa M, Kanamori H, Kawasaki M, Nishigaki K, Takemura G, Kumazaki M, Akao Y, Minatoguchi S (2015) MicroRNA-145 repairs infarcted myocardium by accelerating cardiomyocyte. Autophagy. Am J Physiol Heart Circ Physiol 309:H1813–H182626432843 10.1152/ajpheart.00709.2014
14. Satake A Minatoguchi S Heishima K Yasuda S Murase H Yoshizumi R Komaki M Baba S Ojio S Tanaka T Akao Y Minatoguchi S Okura H An increase in plasma micro RNA-143 in the acute phase is positively correlated with recovery of cardiac function in the chronic phase in patients with acute myocardial infarction Circ J 2023 87 824 833 10.1253/circj.CJ-22-0698 36775328
Satake A, Minatoguchi S, Heishima K, Yasuda S, Murase H, Yoshizumi R, Komaki M, Baba S, Ojio S, Tanaka T, Akao Y, Minatoguchi S, Okura H (2023) An increase in plasma micro RNA-143 in the acute phase is positively correlated with recovery of cardiac function in the chronic phase in patients with acute myocardial infarction. Circ J 87:824–83336775328 10.1253/circj.CJ-22-0698
15. Akao Y Naoe T Hirata I Nakagawa Y Iio A Identification of non-coding RNAs embracing microRNA-143/145 cluster Mol Cancer 2010 9 136 10.1186/1476-4598-9-136 20525177
Akao Y, Naoe T, Hirata I, Nakagawa Y, Iio A (2010) Identification of non-coding RNAs embracing microRNA-143/145 cluster. Mol Cancer 9:13620525177 10.1186/1476-4598-9-136
16. Fukada M Matsuhashi N Takahashi T Sugito N Heishima K Yoshida K Akao Y Postoperative changes in plasma miR21-5p as a novel biomarker for colorectal cancer recurrence: a prospective study Cancer Sci 2021 112 4270 4280 10.1111/cas.15065 34270831
Fukada M, Matsuhashi N, Takahashi T, Sugito N, Heishima K, Yoshida K, Akao Y (2021) Postoperative changes in plasma miR21-5p as a novel biomarker for colorectal cancer recurrence: a prospective study. Cancer Sci 112:4270–428034270831 10.1111/cas.15065
17. Heishima K Mori T Ichikawa Y Sakai H Kuranaga Y Nakagawa T Tanaka Y Okamura Y Masuzawa M Sugito N Murakami M Yamada N Akao Y Maruo K MicroRNA-214 and microRNA-126 are potential biomarkers for malignant endothelial proliferative diseases Int J Mol Sci 2015 16 25377 25391 10.3390/ijms161025377 26512652
Heishima K, Mori T, Ichikawa Y, Sakai H, Kuranaga Y, Nakagawa T, Tanaka Y, Okamura Y, Masuzawa M, Sugito N, Murakami M, Yamada N, Akao Y, Maruo K (2015) MicroRNA-214 and microRNA-126 are potential biomarkers for malignant endothelial proliferative diseases. Int J Mol Sci 16:25377–2539126512652 10.3390/ijms161025377
18. Zhelankin AV Vasiliev SV Stonogina DA Babalyan KA Sharova EI Doludin YV Shchekochikhin DY Generozov EV Akselrod AS Elevated plasma levels of circulating extracellular miR-320a-3p in patients with paroxysmal atrial fibrillation Int J Med Sci 2020 21 3485
Zhelankin AV, Vasiliev SV, Stonogina DA, Babalyan KA, Sharova EI, Doludin YV, Shchekochikhin DY, Generozov EV, Akselrod AS (2020) Elevated plasma levels of circulating extracellular miR-320a-3p in patients with paroxysmal atrial fibrillation. Int J Med Sci 21:3485
19. Yamano T Kubo S Sonoda E Kominato T Kimura K Yasuhara M Kataoka K Son J Babaya A Takenaka Y Matsubara T Beppu N Ikeda M Assessment of circulating microRNA specific for patients with familial adenomatous polyposis PLoS ONE 2021 16 e0250072 10.1371/journal.pone.0250072 33945535
Yamano T, Kubo S, Sonoda E, Kominato T, Kimura K, Yasuhara M, Kataoka K, Son J, Babaya A, Takenaka Y, Matsubara T, Beppu N, Ikeda M (2021) Assessment of circulating microRNA specific for patients with familial adenomatous polyposis. PLoS ONE 16:e025007233945535 10.1371/journal.pone.0250072
20. Boettger T Beetz N Kostin S Schneider J Kruger M Hein L Braun T Acquisition of the contractile phenotype by murine arterial smooth muscle cells depends on the Mir143/145 gene cluster J Clin Invest 2009 119 2634 2647 10.1172/JCI38864 19690389
Boettger T, Beetz N, Kostin S, Schneider J, Kruger M, Hein L, Braun T (2009) Acquisition of the contractile phenotype by murine arterial smooth muscle cells depends on the Mir143/145 gene cluster. J Clin Invest 119:2634–264719690389 10.1172/JCI38864
21. Norata GD Pinna C Zappella F Elia L Sala A Condorelli G、Catapano AL (2012) MicroRNA 143–145 deficiency impairs vascular function Int J Immunopathol Pharmacol 2012 25 467 474 10.1177/039463201202500216 22697078
Norata GD, Pinna C, Zappella F, Elia L, Sala A (2012) Condorelli G、Catapano AL (2012) MicroRNA 143–145 deficiency impairs vascular function. Int J Immunopathol Pharmacol 25:467–47422697078 10.1177/039463201202500216
22. Hergenreider E Heydt S Treguer K Boettger T Horrevoets AJG Zeiher AM Scheffer MP Frangakis AS Yin X Mayr M Braun T Urbich C Boon RA Dimmeler S (2012) Atheroprotective communication between endothelial cells and smooth muscle cells through miRNAs Nat Cell Biol 2012 14 249 256 10.1038/ncb2441 22327366
Hergenreider E, Heydt S, Treguer K, Boettger T, Horrevoets AJG, Zeiher AM, Scheffer MP, Frangakis AS, Yin X, Mayr M, Braun T, Urbich C, Boon RA (2012) Dimmeler S (2012) Atheroprotective communication between endothelial cells and smooth muscle cells through miRNAs. Nat Cell Biol 14:249–25622327366 10.1038/ncb2441
23. Boettger T Beetz N Kostin S Schneider J Kruger M Hein L Braun T Acquisition of the contractile phenotype by murine arterial smooth muscle cells depends on the Mir143/145 gene cluster J Clin Invest 2009 119 2634 2647 10.1172/JCI38864 19690389
Boettger T, Beetz N, Kostin S, Schneider J, Kruger M, Hein L, Braun T (2009) Acquisition of the contractile phenotype by murine arterial smooth muscle cells depends on the Mir143/145 gene cluster. J Clin Invest 119:2634–264719690389 10.1172/JCI38864
24. Fichtcherer S De Rosa S Fox H Schwietz T Fischer A Liebetrau C Weber M Hamm CW Röxe T Müller-Ardogan M Bonauer A Zeiher AM Dimmeler S Circulating microRNAs in patients with coronary artery disease Circ Res 2010 107 677 684 10.1161/CIRCRESAHA.109.215566 20595655
Fichtcherer S, De Rosa S, Fox H, Schwietz T, Fischer A, Liebetrau C, Weber M, Hamm CW, Röxe T, Müller-Ardogan M, Bonauer A, Zeiher AM, Dimmeler S (2010) Circulating microRNAs in patients with coronary artery disease. Circ Res 107:677–68420595655 10.1161/CIRCRESAHA.109.215566
25. Santovito D Mandolini C Marcantonio P De Nardis V Bucci M Paganelli C Magnacca F Ucchino S Mastroiacovo D Desideri G Mezzetti A Cipollone F Overexpression of microRNA-145 in atherosclerotic plaques from hypertensive patients Expert Opin Ther Targets 2013 17 217 223 10.1517/14728222.2013.745512 23339529
Santovito D, Mandolini C, Marcantonio P, De Nardis V, Bucci M, Paganelli C, Magnacca F, Ucchino S, Mastroiacovo D, Desideri G, Mezzetti A, Cipollone F (2013) Overexpression of microRNA-145 in atherosclerotic plaques from hypertensive patients. Expert Opin Ther Targets 17:217–22323339529 10.1517/14728222.2013.745512
26. Elia L Quintavalle M Zhang J Contu R Cossu L Latronico MVG Peterson KL Indolfi C Catalucci D Chen J Courtneidge SA Condorelli G The knockout of miR-143 and -145 alters smooth muscle muscle cell maintenance and vascular homeostasis in mice: correlates with human disease Cell Death Differ 2009 16 1590 1598 10.1038/cdd.2009.153 19816508
Elia L, Quintavalle M, Zhang J, Contu R, Cossu L, Latronico MVG, Peterson KL, Indolfi C, Catalucci D, Chen J, Courtneidge SA, Condorelli G (2009) The knockout of miR-143 and -145 alters smooth muscle muscle cell maintenance and vascular homeostasis in mice: correlates with human disease. Cell Death Differ 16:1590–159819816508 10.1038/cdd.2009.153
27. Geekiyanagea H Rayatpishehb S Wohlschlegelb JA Brown Jrc R Ambrosa V Extracellular microRNAs in human circulation are associated with miRISC complexes that are accessible to anti-AGO2 antibody and can bind target mimic oligonucleotides Proc Natl Acad Sci USA 2020 117 24213 24223 10.1073/pnas.2008323117 32929008
Geekiyanagea H, Rayatpishehb S, Wohlschlegelb JA, Brown Jrc R, Ambrosa V (2020) Extracellular microRNAs in human circulation are associated with miRISC complexes that are accessible to anti-AGO2 antibody and can bind target mimic oligonucleotides. Proc Natl Acad Sci USA 117:24213–2422332929008 10.1073/pnas.2008323117
28. Ribeiro-Rodrigues TM Laundos TL Pereira-Carvalho R Batista-Almeida D Pereira R Coelho-Santos V Silva AP Fernandes R Zurarte M Enguita FJ Costa MC Pinto-do-O P Pinto M Gouveia P Ferreira L Mason JC Pereira P Kwak BR Nascimento DS Girao H Exosomes secreted by cardiomyocytes subjected to ischaemia promote cardiac angiogenesis Cardiovasc Res 2017 113 1338 1350 10.1093/cvr/cvx118 28859292
Ribeiro-Rodrigues TM, Laundos TL, Pereira-Carvalho R, Batista-Almeida D, Pereira R, Coelho-Santos V, Silva AP, Fernandes R, Zurarte M, Enguita FJ, Costa MC, Pinto-do-O P, Pinto M, Gouveia P, Ferreira L, Mason JC, Pereira P, Kwak BR, Nascimento DS, Girao H (2017) Exosomes secreted by cardiomyocytes subjected to ischaemia promote cardiac angiogenesis. Cardiovasc Res 113:1338–135028859292 10.1093/cvr/cvx118
29. Zhao W Zhao SP Zhao YH MicroRNA-143/-145 in cardiovascular diseases BioMed Res Int 2015 10.1155/2015/531740 26881202
Zhao W, Zhao SP, Zhao YH (2015) MicroRNA-143/-145 in cardiovascular diseases. BioMed Res Int. 10.1155/2015/53174026881202 10.1155/2015/531740
30. Mohl W Spitzer E Mader RM Wagh V Nguemo F Milasinovic D Jusic A Khazen C Szodorai E Birkenberg B Lubec G Hescheler J Serruys PW Acute molecular effects of pressure-controlled intermittent coronary sinus occlusion in patients with advanced heart failure ESC Heart Fail 2018 5 1176 1183 10.1002/ehf2.12354 30230713
Mohl W, Spitzer E, Mader RM, Wagh V, Nguemo F, Milasinovic D, Jusic A, Khazen C, Szodorai E, Birkenberg B, Lubec G, Hescheler J, Serruys PW (2018) Acute molecular effects of pressure-controlled intermittent coronary sinus occlusion in patients with advanced heart failure. ESC Heart Fail 5:1176–118330230713 10.1002/ehf2.12354
31. Bhat PK Ashwath ML Rosenbaum DS Costantini O Usefulness of left ventricular end-systolic dimension by echocardiography to predict reverse remodeling in patients with newly diagnosed severe left ventricular systolic dysfunction Am J Cardiol 2012 110 83 87 10.1016/j.amjcard.2012.02.054 22551737
Bhat PK, Ashwath ML, Rosenbaum DS, Costantini O (2012) Usefulness of left ventricular end-systolic dimension by echocardiography to predict reverse remodeling in patients with newly diagnosed severe left ventricular systolic dysfunction. Am J Cardiol 110:83–8722551737 10.1016/j.amjcard.2012.02.054
32. Moilanen AM Rysa J Mustonen E Serpi R Aro J Tokola H Leskinen H Manninen A Levijoki J Vuolteenaho O Ruskoaho H Intramyocardial BNP gene delivery improves cardiac function through distinct context-dependent mechanisms Circ Heart Fail 2011 4 483 495 10.1161/CIRCHEARTFAILURE.110.958033 21558448
Moilanen AM, Rysa J, Mustonen E, Serpi R, Aro J, Tokola H, Leskinen H, Manninen A, Levijoki J, Vuolteenaho O, Ruskoaho H (2011) Intramyocardial BNP gene delivery improves cardiac function through distinct context-dependent mechanisms. Circ Heart Fail 4:483–49521558448 10.1161/CIRCHEARTFAILURE.110.958033
33. McMurray JJV Packer M Desai AS Gong J Lefkowitz MP Rizkala AR Rouleau JL Shi VC Solomon SD Swedberg K Zile MR PARADIGM-HF Investigators and Committees Angiotensin-Neprilysin inhibition versus Enalapril in heart failure N Engl J Med 2014 371 993 1004 10.1056/NEJMoa1409077 25176015
McMurray JJV, Packer M, Desai AS, Gong J, Lefkowitz MP, Rizkala AR, Rouleau JL, Shi VC, Solomon SD, Swedberg K, Zile MR, PARADIGM-HF Investigators and Committees (2014) Angiotensin-Neprilysin inhibition versus Enalapril in heart failure. N Engl J Med 371:993–100425176015 10.1056/NEJMoa1409077
34. Myhre PL Vaduganathan M Claggett B Packer M Desai AS Rouleau JL Zile MR Swedberg K Lefkowitz M Shi V McMurray JJV Solomon SD B-type natriuretic peptide during treatment with sacubitril/valsartan: The PARADIGM-HF Trial J Am Coll Cardiol 2019 73 1264 1272 10.1016/j.jacc.2019.01.018 30846338
Myhre PL, Vaduganathan M, Claggett B, Packer M, Desai AS, Rouleau JL, Zile MR, Swedberg K, Lefkowitz M, Shi V, McMurray JJV, Solomon SD (2019) B-type natriuretic peptide during treatment with sacubitril/valsartan: The PARADIGM-HF Trial. J Am Coll Cardiol 73:1264–127230846338 10.1016/j.jacc.2019.01.018
35. Hergenreider E Heydt S Treguer K Boettger T Horrevoets AJG Zeiher AM Scheffer MP Frangakis AS Yin X Mayr M Braun T Urbich C Boon RA Dimmeler BS Atheroprotective communication between endothelial cells and smooth muscle cells through miRNAs Nat Cell Biol 2012 14 249 256 10.1038/ncb2441 22327366
Hergenreider E, Heydt S, Treguer K, Boettger T, Horrevoets AJG, Zeiher AM, Scheffer MP, Frangakis AS, Yin X, Mayr M, Braun T, Urbich C, Boon RA, Dimmeler BS (2012) Atheroprotective communication between endothelial cells and smooth muscle cells through miRNAs. Nat Cell Biol 14:249–25622327366 10.1038/ncb2441
36. Kia M Cooley A Rimmer G MacDonald T Barber K Manion P Shapiro B Socey J Iddings D The efficacy of B-type natriuretic peptide for early identification of blood loss in traumatic injury Am J Surg 2006 191 353 357 10.1016/j.amjsurg.2005.10.033 16490546
Kia M, Cooley A, Rimmer G, MacDonald T, Barber K, Manion P, Shapiro B, Socey J, Iddings D (2006) The efficacy of B-type natriuretic peptide for early identification of blood loss in traumatic injury. Am J Surg 191:353–35716490546 10.1016/j.amjsurg.2005.10.033
37. Wang J Tong KS Wong LL Liew OW Raghuram D Richards AM Chen YT MicroRNA-143 modulates the expression of natriuretic peptide receptor 3 in cardiac cells Sci Rep 2018 8 7055 10.1038/s41598-018-25489-3 29728596
Wang J, Tong KS, Wong LL, Liew OW, Raghuram D, Richards AM, Chen YT (2018) MicroRNA-143 modulates the expression of natriuretic peptide receptor 3 in cardiac cells. Sci Rep 8:705529728596 10.1038/s41598-018-25489-3
38. Weidemann A Klanke B Wagner M Volk T William C Wiesener MS Eckardt KU Warnecke C Hypoxia, via stabilization of the hypoxia-inducible factor HIF-1 alpha, is a direct and sufficient stimulus for brain-type natriuretic peptide induction Biochem J 2008 409 233 242 10.1042/BJ20070629 17822384
Weidemann A, Klanke B, Wagner M, Volk T, William C, Wiesener MS, Eckardt KU, Warnecke C (2008) Hypoxia, via stabilization of the hypoxia-inducible factor HIF-1 alpha, is a direct and sufficient stimulus for brain-type natriuretic peptide induction. Biochem J 409:233–24217822384 10.1042/BJ20070629
39. Chen G Wang M Ruan Z Zhu L Tang C Mesenchymal stem cell-derived exosomal miR-143-3p suppresses myocardial ischemia-reperfusion injury by regulating autophagy Life Sci 2021 280 119742 10.1016/j.lfs.2021.119742 34166712
Chen G, Wang M, Ruan Z, Zhu L, Tang C (2021) Mesenchymal stem cell-derived exosomal miR-143-3p suppresses myocardial ischemia-reperfusion injury by regulating autophagy. Life Sci 280:11974234166712 10.1016/j.lfs.2021.119742
