Cardiology Research, ISSN 1923-2829 print, 1923-2837 online, Open Access
Article copyright, the authors; Journal compilation copyright, Cardiol Res and Elmer Press Inc
Journal website https://www.cardiologyres.org

Original Article

Volume 15, Number 2, April 2024, pages 108-116

Increased Cyclic Guanosine Monophosphate and Interleukin-1Beta Is Activated by Mitochondrial Dysfunction and Associated With Heart Failure in Atrial Fibrillation Patients

Figures

Figure 1.
Figure 1. Comparison of serum 2'3'-cGAMP and interleukin (IL)-1β in AF patients with and without HF. Differences in serum levels of 2'3'-cGAMP (a) and IL-1β (b) between two groups were performed with Student’s t-test. *P < 0.05. 2'3'-cGAMP: 2'3'-cyclic GMP-AMP; GMP: guanosine monophosphate; AMP: adenosine monophosphate; HF: heart failure; AF: atrial fibrillation.
Figure 2.
Figure 2. Complex I inhibitor rotenone induced mitochondrial dysfunction in neonatal rat ventricular cardiomyocytes (NRVMs). (a) NRVMs were treated with GAS antagonist RU.521 (10 µM) for 24 h in the presence of rotenone (0.1 µM). ATP content was measured using colorimetric tests. (b) Using immunofluorescence, cytoplasmic calcium in NRVMs was detected by Rhod-2 probe (labeled in red). Scale bar = 50 µm. * P < 0.05. GAS: guanosine monophosphate (GMP)-adenosine monophosphate (AMP) synthase; ATP: adenosine triphosphate.
Figure 3.
Figure 3. Rotenone inhibited mitochondrial dysfunction-induced cGAS-STING pathway in neonatal rat ventricular cardiomyocytes (NRVMs). (a) NRVMs were treated with GAS antagonist RU.521 (10 µM) for 24 h in the presence of rotenone (0.1 µM). The content of 2'3'-cGAMP in NRVMs was measured using ELISA kit. (b) Western blot analysis showed the expression of cGAS and STING phosphorylation. (c) Quantification of the expression of cGAS and STING phosphorylation evaluated by western blots. *P < 0.05. ELISA: enzyme-linked immunoassay; cGAS-STING: cyclic guanosine monophosphate (GMP)-adenosine monophosphate (AMP) synthase-stimulator interferon genes; 2'3'-cGAMP: 2'3'-cyclic GMP-AMP.
Figure 4.
Figure 4. Inhibition of cGAS-STING attenuated the activation of NLRP3 inflammasome in neonatal rat ventricular cardiomyocytes (NRVMs). (a) NRVMs were treated with GAS antagonist RU.521 (10 µM) for 24 h in the presence of rotenone (0.1 µM). Western blot analysis showed the expression of NLRP3 and the cleavage of GSDMD. (b) The supernatant was collected and the concentration of interleukin (IL)-1β and IL-18 was measured using ELISA kit. *P < 0.05. NLRP3: nod-like receptor protein 3; ELISA: enzyme-linked immunoassay; cGAS-STING: cyclic guanosine monophosphate (GMP)-adenosine monophosphate (AMP) synthase-stimulator interferon genes; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GSDMD: gasdermin D.

Tables

Table 1. Baseline Characteristics of Enrollments
 
No HF during follow-up (n = 61)HFpEF/HFmrEF during follow-up (n = 13)HFrEF during follow-up (n = 32)P value
*P < 0.05. Data are expressed as mean ± SD or n (%). Antiarrhythmic drugs include amiodarone and propafenone. ACEI/ARB/ARNI: angiotensin-converting enzyme inhibitor/angiotensin receptor II blockade/angiotensin receptor neprilysin inhibitor; AF: atrial fibrillation; CAD: coronary artery disease; LAD: left atrial diameter; LVEF: left ventricular ejection fraction; MRA: mineralocorticoid receptor antagonist; NT-proBNP: N-terminal fragment B-type natriuretic peptide; HF: heart failure; HFrEF: HF with reduced ejection fraction; HFmrEF: HF with mid-range ejection fraction; HFpEF: HF with preserved ejection fraction.
Age (years)64.46 ± 10.9467.23 ± 11.4364.88 ± 9.710.70
Male, n (%)16 (26.2)4 (30.8)16 (50.0)0.07
Persistent/long-standing persistent AF, n (%)37 (60.7)/24 (39.3)6 (46.2)/7 (53.8)13 (40.6)/19 (59.4)0.16
Diabetes, n (%)11 (18.0)2 (15.4)6 (18.8)0.07
Smoking, n (%)16 (26.2)4 (30.8)16 (50.0)0.07
Hypertension, n (%)40 (65.6)9 (69.2)20 (62.5)0.91
CAD, n (%)15 (24.6)6 (46.2)11 (34.4)0.25
Stroke, n (%)15 (24.6)5 (38.5)9 (28.1)0.59
Anticoagulant, n (%)29 (47.5)12 (92.3)24 (75.0)< 0.01*
ACEI/ARB/ARNI, n (%)37 (60.7)8 (61.5)13 (40.6)0.16
β-blocker, n (%)29 (47.5)6 (46.2)17 (50.0)0.86
MRA, n (%)5 (8.2)0 (0.0)2 (6.3)0.56
Antiarrhythmic drugs, n (%)24 (39.3)8 (61.5)10 (31.3)0.17
LAD (mm)45.93 ± 5.2946.23 ± 3.8648.44 ± 6.040.10
LVEF (%)57.03 ± 9.3650.31 ± 10.5633.78 ± 2.97< 0.01*
NT-proBNP (pg/mL)143.72 ± 90.10662.87 ± 98.611,099.46 ± 107.67< 0.01*

 

Table 2. Multivariate Logistic Analysis for the Incidence of HF in AF Patients
 
VariablesOR (95% CI)P value
*P < 0.05. HF: heart failure; ACEI/ARB/ARNI: angiotensin-converting enzyme inhibitor/angiotensin receptor II blockade/angiotensin receptor neprilysin inhibitor; AF: atrial fibrillation; CAD: coronary artery disease; LAD: left atrial diameter; LVEF: left ventricular ejection fraction; MRA: mineralocorticoid receptor antagonist; OR: odds ratio; CI: confidence interval; IL: interleukin; 2'3'-cGAMP: 2'3'-cyclic GMP-AMP; GMP: guanosine monophosphate; AMP: adenosine monophosphate.
Male0.93 (0.17, 8.14)0.92
Type of AF7.01 (0.85, 56.37)0.47
Smoking0.92 (0.17, 3.80)0.54
Hypertension1.85 (0.58, 3.92)0.64
Diabetes1.94 (0.42, 7.23)0.83
CAD1.72 (0.26, 3.47)0.91
Stroke1.88 (0.77, 4.28)0.28
Anticoagulant2.86 (1.54, 4.21)0.02*
ACEI/ARB/ARNI0.49 (0.13, 3.42)0.27
β-blocker1.37 (0.54, 2.09)0.53
MRA1.41 (0.75, 3.03)0.42
Antiarrhythmic drugs1.55 (0.71, 4.08)0.37
LAD1.32 (0.56, 3.93)0.68
2'3'-cGAMP1.74 (1.34, 2.85)0.02*
IL-1β2.59 (1.83, 4.25)< 0.01*