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Apelin‑13 in the paraventricular nucleus (PVN) attenuates myocardial ischemia through V1a receptors in PVN/nucleus tractus solitarii (NTS) and GARγ2 in NTS

  • Authors:
    • Wen Yan
    • Dan Wang
    • Xinmin Zhang
    • Chengluan Xuan
  • View Affiliations / Copyright

    Affiliations: Department of Anesthesia, The Second Hospital of Jilin University, Changchun, Jilin 130021, P.R. China, Department of Anesthesia, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China
    Copyright: © Yan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 211
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    Published online on: September 30, 2025
       https://doi.org/10.3892/ijmm.2025.5652
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Abstract

The apelin system plays a significant role in central blood pressure regulation, but its role in the neural control of myocardial protection remains poorly understood. The present study evaluated the effects of apelin‑13 in the paraventricular nucleus (PVN) on myocardial infarction (MI). In a male rat MI model, apelin‑13 expression was decreased in PVN, while Vasopressin 1a (V1a) receptor expression was increased in both PVN and nucleus tractus solitarii (NTS) and GABAA receptor (GAR)γ2 expression was increased in NTS. Cardiac function was assessed after microinjection of apelin‑13 or gene transfer of apelin‑13 into the PVN. Apelin‑13 overexpression in PVN markedly improved MI cardiac function, as evidenced by left ventricular end‑diastolic diameter, left ventricular end‑systolic diameter, left ventricular ejection fraction and left ventricular fractional shortening, along with decreased plasma noradrenaline and increased vasopressin levels. Mechanistically, both TGF‑β/Smad signaling and Bax/Bcl‑2 expression were implicated in heart tissue. Additionally, serum levels of four parasympathetic neuropeptides (somatostatin, cholecystokinin, glucagon‑like peptide‑1 and vasoactive intestinal peptide) were elevated in parallel with cardiac function improvement. Notably, V1a receptor antagonist administration in PVN/NTS or GAR agonist treatment in NTS attenuated the cardioprotective effects of apelin‑13. These findings demonstrated that PVN apelin‑13 overexpression improves cardiac function through V1a receptors (PVN/NTS) and GARγ2 (NTS), involving both parasympathetic neuroendocrine activation and modulation of myocardial apoptotic/inflammatory pathways. The present study provided novel insights into neural mechanisms of cardiovascular regulation.
View Figures

Figure 1

Expression of apelin-13, the V1a
receptor and the GARγ2 receptor in MI model rats. (A)
Representative western blots of apelin-13, the V1a receptor, the
GBR1 and the GARγ2 in the PVN, NTS and RVLM from MI model rats and
sham-operated control rats. (B) Quantification of apelin-13 protein
levels. (C) mRNA expression levels of apelin-13. (D) Quantification
of V1a receptor protein levels. (E) mRNA expression levels of the
V1a receptor. (F) Quantification of GBR1 protein levels. (G) mRNA
expression levels of GBR1. (H) Quantification of GARγ2 protein
levels. (I) mRNA expression levels of GARγ2. Normality was tested
using the Shapiro-Wilk test. The data (n=6 rats per group; 3
technical replicates) in B-I are shown as mean ± SEM and were
analyzed using an unpaired two-tailed t test.
*P<0.05; **P<0.01;
***P<0.001. Post-hoc power exceeded 80% for all
comparisons. V1a, Vasopressin 1a; GAR, GABAA receptor;
MI, myocardial infarction; GBR1, GABAB receptor1; PVN,
paraventricular nucleus; NTS, nucleus tractus solitarii; RVLM,
rostral ventrolateral medulla.

Figure 2

Effects of apelin-13 microinjection
into the PVN on cardiac function in MI model rats. (A) MI models
were established one day after implanting an osmotic mini-pump
delivering apelin-13 into the PVN. Cardiac function was assessed
and tissue samples were collected 28 days after mini-pump
implantation. (B) Representative western blots for the V1a receptor
and GARγ2 in the PVN and NTS from MI model rats and MI model rats
continuously microinjected with apelin-13 into the PVN for 28 days.
(C) Quantification of V1a receptor protein levels. (D)
Quantification of GARγ2 receptor protein levels. (E) Representative
ultrasound images (M-mode) performed on days 28 after MI and
sham-operated control. Echocardiographic results for (F) LVEDD, (G)
LVESD, (H) LVEF and (I) LVFS (n=6 in the sham-operated control
group; n=5 in the MI group, one rat succumbed at 20 days; n=6 in
the MI + apelin group). Plasma levels of (J) SST, (K) CCK, (L) VIP,
(M) GLP-1, (N) noradrenaline and (O) vasopressin (n=6 in the
sham-operated control group; n=5 in the MI group, one rat succumbed
at 20 days; n=6 in the MI + apelin group). Normality was tested
using the Shapiro-Wilk test. The data in C-D show mean ± SEM; F-O
show mean ± SD. Statistical significance was determined by one-way
ANOVA, followed by Tukey's multiple-comparisons test.
*P<0.05; **P<0.01;
***P<0.001. Post-hoc power exceeded 80% for all
comparisons. PVN, paraventricular nucleus; MI, myocardial
infarction; V1a, Vasopressin 1a; GAR, GABAA receptor;
NTS, nucleus tractus solitarii; LVEDD, left ventricular
end-diastolic diameter; LVESD, lower left ventricular end-systolic
diameter; LVEF, left ventricular ejection fraction; LVFS, left
ventricular fraction shortening; SST, isomatostatin; CCK,
cholecystokinin; VIP, vasoactive intestinal peptide; GLP-1,
glucagon-like peptide 1.

Figure 3

Effects of continuous apelin-13
infusion into the PVN on myocardial ischemia-associated apoptotic
and inflammatory protein expression in MI rats. (A) Representative
image and quantification of TGF-β1+ cells. (B)
Representative image and expression index of Smad2. (C)
Representative image and optical density of Bax. (D) Representative
image and quantification of Bcl-2+ cells. (E)
Representative multiplex fluorescent immunohistochemistry staining
images and relative fluorescence intensity of TGF-β1, Smad2, Bax
and Bcl-2 (n=6 in the sham-operated control group; n=5 in the MI
group, one rat succumbed at 20 days; n=6 in the MI + apelin group).
Normality was tested using the Shapiro-Wilk test. The data are
shown as mean ± SD and were analyzed using one-way ANOVA, followed
by Tukey's multiple-comparisons test. *P<0.05;
**P<0.01; ***P<0.001. Post-hoc power
exceeded 80% for all key comparisons. PVN, paraventricular nucleus;
MI, myocardial infarction.

Figure 4

Comparison of effects between
apelin-13 gene transfer and continuous infusion approaches. (Aa)
After implanting apelin-13 osmotic mini-pumps or delivering
AAV2-apelin-13 to the PVN, MI was induced and heart and brain
tissues collected at 3-, 7-, 14- and 28-days post-MI (n=6 in each
group). Location of the stained PVN brain sections shown in (b) and
(c), based on the rat brain atlas of C. Watson and G. Paxinos
(24). (c) Fluorescence
micrograph (magnification, ×10) demonstrating the localization of
apelin-13 within the PVN. 3v is the third ventricle. (d) Overlap of
(e), (f) and (g), showing that the green fluorescence is neuronally
located. (e) Fluorescence micrograph (magnification, ×40)
indicating the localization of apelin-13 in PVN cells immunostained
with an anti-apelin antibody (green). (f) Same field of cells as in
panel (e), immunostained with anti-NeuN antibodies (red). (g) DAPI
staining of the same field of cells in panel (e). (B) Molecular
pathways regulating apoptotic and inflammatory pathways were
assessed through western blotting of the expression of Bax, Bcl-2,
TGF-β1 and Smad2 over time in MI model rats and the expression of
the V1a receptor and GARγ2 induced by apelin-13 overexpression in
the PVN. (C and I) Quantification of the fold change in target gene
expression vs. GAPDH expression. The data (n=6 rats per group, 3
technical replicates) in (C-I) are shown as mean ± SEM and were
analyzed via an unpaired two-tailed t test. *P<0.05;
**P<0.01; ***P<0.001. Post-hoc power
exceeded 80% for all key comparisons. PVN, paraventricular nucleus;
MI, myocardial infarction; GT, gene transfer; CI, continuous
infusion.

Figure 5

Effects of the APJ receptor and the
GARγ2 receptor on the V1a receptor in primary cultured neurons. (A)
Immunofluorescence of F13A-, muscimol- and control-treated primary
neurons. Neurons were treated for 12, 24, 48, or 72 h. (B)
Representative positive fluorescence. (C and D) Representative
western blot image of the V1a receptor in primary cultured neurons
treated with F13A and muscimol and quantification of V1a receptor
expression. (E) mRNA expression levels of the V1a receptor in
neurons treated with F13A and muscimol. The data (n=5 experiments)
in (B) are shown as mean ± SD and were analyzed via one-way ANOVA,
followed by Dunnett's multiple-comparisons test. Normality was
tested using the Shapiro-Wilk test. The data (n=5; 3 technical
replicates) in (D and E) are shown as mean ± SEM and were analyzed
via one-way ANOVA, followed by Dunnett's multiple-comparisons test.
*P<0.05; **P<0.01; and
***P<0.001vs. the control group. Post-hoc power
exceeded 80% for all key comparisons. APJ, orphan
G-protein-coupled; GAR, GABAA receptor; V1a, Vasopressin
1a.

Figure 6

Effect of apelin-13 overexpression
(AAV2-apelin-13 gene transfer) in the PVN on V1a receptor-mediated
improvements in cardiac function and cardiac morphology in MI model
rats. (A) The MI model was constructed one day following the
transfer of the apelin-13 gene into PVN and the implantation of
mini-pump (7 rats in each group). Echocardiographic results for (B)
LVEF, (C) LVSF, (D) LVEDD and (E) LVESD. (F) Representative
Masson's trichrome staining and (I) quantification. (G)
Representative TUNEL staining and (J) quantification of TUNEL
staining. (H) Representative TTC staining and (K) quantification.
The black scale bar in F is 100 μm; the white scale bar in G
is 50 μm; and the black scale bar in I is 1 cm. At 28 days
after MI modelling, the sham-operated control group had 7 rats, the
MI group had 5 rats (2 rats succumbed), the MI + apelin-13 group
had 7 rats, the MI + apelin-13 + SR49059 (PVN) group had 5 rats (2
rats succumbed), the MI + apelin-13 + SR49059 (NTS) group had 6
rats (1 rat succumbed) and the MI + apelin-13 + muscimol (NTS)
group had 6 rats (1 rat succumbed). Normality was tested using the
Shapiro-Wilk test. The data in B-E and H-M are shown as means ± SDs
and were analyzed using one-way ANOVA, followed by Tukey's
multiple-comparisons test. In B-E and H-K, *P<0.05
vs. sham-operated control group; †P<0.05 vs. MI
group; ‡P<0.05 vs. MI + apelin-13 group;
§P<0.05 vs. MI + apelin-13 + SR49059(PVN);
#P<0.05 vs. MI + apelin-13 + SR49059 (NTS). The (L)
MAP and (M) heart rate were recorded in a conscious state at days
3, 7, 14 and 28 after MI modelling. *P<0.05;
**P<0.01; ***P<0.001; and ns, not
significant, vs. the sham-operated control group. Post-hoc power
exceeded 80% for all key comparisons. PVN, paraventricular nucleus;
V1a, Vasopressin 1a; MI, myocardial infarction; ECG,
Echocardiographic; LVEF, left ventricular ejection fraction; LVFS,
left ventricular fraction shortening; LVEDD, left ventricular
end-diastolic diameter; LVESD, lower left ventricular end-systolic
diameter; NTS, nucleus tractus solitarii; MAP, mean arterial
pressure.

Figure 7

Mechanisms of the effect of apelin-13
overexpression (AAV2-apelin-13 gene transfer) in the PVN on V1a
receptor-mediated improvement in cardiac function in MI model rats.
(A) Representative western blot image of V1a receptor expression in
the PVN or NTS and GARγ2 expression in the NTS. (B-D)
Quantification of V1a receptor expression. (E) Representative
western blotting to assess Bcl-2, Bax, TGF-β1 and Smad2 protein
levels in the heart and (F-I) quantification of the protein levels.
Plasma levels of (J) SST, (K) CCK, (L) VIP and (M) GLP-1 (n=7 in
the sham-operated control group; n=5 in the MI group, 2 rats
succumbed; n=7 in the MI + apelin-13 group; n=5 in the MI +
apelin-13 + SR49059 (PVN) group, 2 rats succumbed; n=6 in the MI +
apelin-13 + SR49059 (NTS) group, 1 rat succumbed; n=6 in the MI +
apelin-13 + muscimol (NTS) group, 1 rat succumbed). Representative
plasma levels of (N) noradrenaline and (O) vasopressin (n=7 in the
sham-operated control group; n=6 in the MI group, 1 rat succumbed;
n=7 in the MI + apelin-13 group; n=6 in the MI + apelin-13 +
SR49059 (PVN) group, 1 rat succumbed; n=6 in the MI + apelin-13 +
SR49059 (NTS) group, 1 rat succumbed; n=7 in the MI + apelin-13 +
muscimol (NTS) group). Normality was tested using the Shapiro-Wilk
test. The data in B-D and F-I show mean ± SEM; data in J-O are
shown as mean ± SD and were analyzed via one-way ANOVA, followed by
Tukey's multiple-comparisons test. In B-D and F-O,
*P<0.05 vs. sham-operated control group;
†P<0.05 vs. MI group; ‡P<0.05 vs. MI +
apelin-13 group; §P<0.05 vs. MI + apelin-13 + SR49059
(PVN); #P<0.05 vs. MI + apelin-13 + SR49059 (NTS).
Post-hoc power exceeded 80% for all key comparisons. PVN,
paraventricular nucleus; V1a, Vasopressin 1a; MI, myocardial
infarction; NTS, nucleus tractus solitarii; GAR, GABAA
receptor; SST, isomatostatin; CCK, cholecystokinin; VIP, vasoactive
intestinal peptide; GLP-1, glucagon-like peptide 1.

Figure 8

Graphical model summarizing the
hypothesized pathway. Overexpression of apelin-13 in the PVN
promotes upregulation of V1a receptors in both the PVN and NTS,
while downregulating GARγ2 in the NTS. These neural modifications
collectively enhance parasympathetic outflow. Furthermore,
paraventricular endocrine system secretes circulating neurohumoral
factors that systemically mediate cardioprotection. PVN,
paraventricular nucleus; V1a, Vasopressin 1a; MI, myocardial
infarction; NTS, nucleus tractus solitarii; GAR, GABAA
receptor; SST, isomatostatin; CCK, cholecystokinin; GLP-1,
glucagon-like peptide 1; VIP, vasoactive intestinal peptide.
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Copy and paste a formatted citation
Spandidos Publications style
Yan W, Wang D, Zhang X and Xuan C: Apelin‑13 in the paraventricular nucleus (PVN) attenuates myocardial ischemia through V1a receptors in PVN/nucleus tractus solitarii (NTS) and GAR&gamma;2 in NTS. Int J Mol Med 56: 211, 2025.
APA
Yan, W., Wang, D., Zhang, X., & Xuan, C. (2025). Apelin‑13 in the paraventricular nucleus (PVN) attenuates myocardial ischemia through V1a receptors in PVN/nucleus tractus solitarii (NTS) and GAR&gamma;2 in NTS. International Journal of Molecular Medicine, 56, 211. https://doi.org/10.3892/ijmm.2025.5652
MLA
Yan, W., Wang, D., Zhang, X., Xuan, C."Apelin‑13 in the paraventricular nucleus (PVN) attenuates myocardial ischemia through V1a receptors in PVN/nucleus tractus solitarii (NTS) and GAR&gamma;2 in NTS". International Journal of Molecular Medicine 56.6 (2025): 211.
Chicago
Yan, W., Wang, D., Zhang, X., Xuan, C."Apelin‑13 in the paraventricular nucleus (PVN) attenuates myocardial ischemia through V1a receptors in PVN/nucleus tractus solitarii (NTS) and GAR&gamma;2 in NTS". International Journal of Molecular Medicine 56, no. 6 (2025): 211. https://doi.org/10.3892/ijmm.2025.5652
Copy and paste a formatted citation
x
Spandidos Publications style
Yan W, Wang D, Zhang X and Xuan C: Apelin‑13 in the paraventricular nucleus (PVN) attenuates myocardial ischemia through V1a receptors in PVN/nucleus tractus solitarii (NTS) and GAR&gamma;2 in NTS. Int J Mol Med 56: 211, 2025.
APA
Yan, W., Wang, D., Zhang, X., & Xuan, C. (2025). Apelin‑13 in the paraventricular nucleus (PVN) attenuates myocardial ischemia through V1a receptors in PVN/nucleus tractus solitarii (NTS) and GAR&gamma;2 in NTS. International Journal of Molecular Medicine, 56, 211. https://doi.org/10.3892/ijmm.2025.5652
MLA
Yan, W., Wang, D., Zhang, X., Xuan, C."Apelin‑13 in the paraventricular nucleus (PVN) attenuates myocardial ischemia through V1a receptors in PVN/nucleus tractus solitarii (NTS) and GAR&gamma;2 in NTS". International Journal of Molecular Medicine 56.6 (2025): 211.
Chicago
Yan, W., Wang, D., Zhang, X., Xuan, C."Apelin‑13 in the paraventricular nucleus (PVN) attenuates myocardial ischemia through V1a receptors in PVN/nucleus tractus solitarii (NTS) and GAR&gamma;2 in NTS". International Journal of Molecular Medicine 56, no. 6 (2025): 211. https://doi.org/10.3892/ijmm.2025.5652
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