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Compensatory activation of PI3K/AKT signaling and apoptosis-associated protein expression in pulmonary hypertension‑induced right ventricular dysfunction

  • Authors:
    • Han Zhang
    • Qixu Zhao
    • Jiashu Sun
    • Yusong Deng
    • Guofeng Xing
    • Weiran Duan
    • Gang Li
    • Qiang Wang
  • View Affiliations / Copyright

    Affiliations: Department of Pediatric Cardiology Surgery, Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, P.R. China, Department of Pediatric Cardiology Surgery, Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, P.R. China
    Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 347
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    Published online on: October 23, 2026
       https://doi.org/10.3892/etm.2026.13342
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Abstract

Pulmonary hypertension (PH) is a progressive cardiopulmonary disorder marked by increased pulmonary vascular resistance and poor prognosis. Right ventricular (RV) dysfunction is the principal determinant of mortality in patients with PH; however, the role of the PI3K/AKT pathway in PH‑induced RV remodeling remains poorly defined. The present study generated a hypobaric hypoxia‑induced PH model in C57BL/6J mice. Echocardiography, histology, reverse transcription‑quantitative PCR, western blotting and immunofluorescence staining were employed to assess RV structure, apoptosis‑associated protein expression and fibrosis, while RNA sequencing was used to identify enriched signaling pathways. In parallel, H9C2 cardiomyocytes were treated with angiotensin II (Ang II) with or without the PI3K inhibitor LY294002 to evaluate apoptosis‑associated protein expression changes. PH model mice developed RV dysfunction characterized by increased Bax and decreased Bcl‑2 expression, accompanied by fibrosis. Transcriptomic analysis revealed significant enrichment of the PI3K/AKT pathway in RV tissues from PH model mice compared with control mice. Pharmacological inhibition of PI3K/AKT signaling with LY294002 further upregulated Bax and downregulated Bcl‑2 in Ang II‑treated H9C2 cells. In conclusion, PI3K/AKT pathway enrichment may represent a compensatory but insufficient response during PH‑induced RV remodeling, and its precise modulation may offer a potential therapeutic strategy for PH‑associated RV failure.
View Figures

Figure 1

Echocardiographic assessment of
cardiac function in PH model mice. (A) Representative M-mode
echocardiographic images of the left ventricle from normoxic C mice
and PH model mice after 28 days of hypobaric hypoxia exposure.
Parameters were measured by echocardiography, including (B) IVSd,
(C) IVSs, (D) LVPWd, (E) LVPWs, (F) LVIDd, (G) LVIDs, (H) LVEDV,
(I) LVESV, (J) FS, (K) SV, (L) LVEF and (M) LVd mass. Data are
presented as mean ± SD (n=8 per group). *P<0.05,
**P<0.01 and ***P<0.001 vs. C group. C,
control; PH, pulmonary hypertension; IVSd, interventricular septal
thickness at diastole; IVSs, interventricular septal thickness at
systole; LVPWd, left ventricular posterior wall thickness at
diastole; LVPWs, left ventricular posterior wall thickness at
systole; LVIDd, left ventricular internal diameter at diastole;
LVIDs, left ventricular internal diameter at systole; LVEDV, left
ventricular end-diastolic volume; LVESV, left ventricular
end-systolic volume; FS, fractional shortening; SV, stroke volume;
LVEF, left ventricular ejection fraction; LVd, left ventricular
diastolic.

Figure 2

Transcriptomic profiling of RV tissue
from PH mode mice. (A) Volcano plot showing differentially
expressed genes between control and PH RV tissues. Red dots
indicate significantly upregulated genes, green dots indicate
significantly downregulated genes (|log2FC|≥1;
FDR<0.05) and blue dots represent genes with no significant
changes. The dashed vertical line indicates log2FC=0,
whereas differentially expressed genes were defined using
|log2FC|≥1 and FDR<0.05; the dashed horizontal line
indicates -log10(FDR)=1.3. (B) Gene Ontology enrichment
analysis of differentially expressed genes. (C) Kyoto Encyclopedia
of Genes and Genomes pathway enrichment analysis. (D) Gene Set
Enrichment Analysis supporting increased activation of the PI3K/AKT
signaling pathway in PH model mice. RV, right ventricular; PH,
pulmonary hypertension; FC, fold change; FDR, false discovery
rate.

Figure 3

Cardiomyocyte apoptosis and RV
fibrosis in PH model mice. (A) Representative western blotting
images showing the protein expression of Bax (21 kDa) and Bcl-2 (26
kDa) in RV tissues from C and PH model mice; GAPDH (36 kDa) was
used as a reference protein. The protein expression levels of (B)
Bax and (C) Bcl-2 were semi-quantified. The relative mRNA
expression levels of (D) Bax and (E) Bcl-2 were
determined by RT-qPCR and normalized to GAPDH. C-1/C-2 and
PH-1/PH-2 denote individual biological replicates from C and PH
groups. The bands shown are representative images from 6 biological
replicates per group; the samples were divided across three
separate gels due to loading capacity. (F) Representative Masson's
trichrome-stained myocardial sections and (G) quantitative analysis
of collagen deposition (blue staining) in RV tissues. Arrows
indicate collagen-positive areas (blue staining). Representative
immunofluorescence images of (H) Bax and (I) Bcl-2 protein levels
in mouse myocardial tissues. Quantitative analysis of (J)
Bax-positive and (K) Bcl-2-positive cell ratios. Data are presented
as the mean ± SD (n=6 per group for western blotting, RT-qPCR and
histology). *P<0.05 vs. C group. RV, right
ventricular; C, control; PH, pulmonary hypertension; RT-qPCR,
reverse transcription-quantitative PCR.

Figure 4

Ki-67-positive cardiomyocytes in the
RV tissues of PH model mice. (A) Representative images of
immunofluorescence staining for Ki-67 (green) in RV myocardial
tissues from C and PH model mice. Cardiomyocytes were identified
based on their characteristic morphology and their nuclear
morphology using DAPI (blue) counterstaining. White arrows indicate
Ki-67-positive cardiomyocytes (Ki-67+/DAPI+
cells with cardiomyocyte morphology). (B) Quantitative analysis of
Ki-67-positive cardiomyocytes, expressed as the percentage of
Ki-67-positive cells relative to total DAPI-positive nuclei. Data
are presented as the mean ± SD of all biological replicates per
group (C and PH), with individual data points overlaid (n=3
biological replicates per group). C-1 to C-3 and PH-1 to PH-3
denote individual biological replicates from C and PH groups.
*P<0.05. RV, right ventricular; C, control; PH,
pulmonary hypertension.

Figure 5

Effects of PI3K inhibition on Ang
II-induced apoptosis in H9C2 cells. (A) Representative western
blotting images and semi-quantitative densitometric analysis of (B)
Bax (21 kDa) and (C) Bcl-2 (26 kDa) protein expression in H9C2
cells treated with Ang II (1 µM; 24 h) in the presence or absence
of the PI3K inhibitor LY294002 (10 µM; 30 min pretreatment). GAPDH
(36 kDa) was used as a reference protein. The relative mRNA
expression levels of (D) Bax and (E) Bcl-2 were
determined by reverse transcription-quantitative PCR and normalized
to GAPDH. Data are presented as the mean ± SD (n=6
biological replicates per group). The western blot bands shown are
representative images from the 6 biological replicates; C-1 and C-2
denote representative untreated C replicates; Ang II-1 and Ang II-2
denote representative Ang II-treated replicates; Ang II +
LY294002-1 and Ang II + LY294002-2 denote representative replicates
treated with both Ang II and LY294002. *P<0.05. Ang
II, angiotensin II; C, control.
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Spandidos Publications style
Zhang H, Zhao Q, Sun J, Deng Y, Xing G, Duan W, Li G and Wang Q: Compensatory activation of PI3K/AKT signaling and apoptosis-associated protein expression in pulmonary hypertension‑induced right ventricular dysfunction. Exp Ther Med 32: 347, 2026.
APA
Zhang, H., Zhao, Q., Sun, J., Deng, Y., Xing, G., Duan, W. ... Wang, Q. (2026). Compensatory activation of PI3K/AKT signaling and apoptosis-associated protein expression in pulmonary hypertension‑induced right ventricular dysfunction. Experimental and Therapeutic Medicine, 32, 347. https://doi.org/10.3892/etm.2026.13342
MLA
Zhang, H., Zhao, Q., Sun, J., Deng, Y., Xing, G., Duan, W., Li, G., Wang, Q."Compensatory activation of PI3K/AKT signaling and apoptosis-associated protein expression in pulmonary hypertension‑induced right ventricular dysfunction". Experimental and Therapeutic Medicine 32.6 (2026): 347.
Chicago
Zhang, H., Zhao, Q., Sun, J., Deng, Y., Xing, G., Duan, W., Li, G., Wang, Q."Compensatory activation of PI3K/AKT signaling and apoptosis-associated protein expression in pulmonary hypertension‑induced right ventricular dysfunction". Experimental and Therapeutic Medicine 32, no. 6 (2026): 347. https://doi.org/10.3892/etm.2026.13342
Copy and paste a formatted citation
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Spandidos Publications style
Zhang H, Zhao Q, Sun J, Deng Y, Xing G, Duan W, Li G and Wang Q: Compensatory activation of PI3K/AKT signaling and apoptosis-associated protein expression in pulmonary hypertension‑induced right ventricular dysfunction. Exp Ther Med 32: 347, 2026.
APA
Zhang, H., Zhao, Q., Sun, J., Deng, Y., Xing, G., Duan, W. ... Wang, Q. (2026). Compensatory activation of PI3K/AKT signaling and apoptosis-associated protein expression in pulmonary hypertension‑induced right ventricular dysfunction. Experimental and Therapeutic Medicine, 32, 347. https://doi.org/10.3892/etm.2026.13342
MLA
Zhang, H., Zhao, Q., Sun, J., Deng, Y., Xing, G., Duan, W., Li, G., Wang, Q."Compensatory activation of PI3K/AKT signaling and apoptosis-associated protein expression in pulmonary hypertension‑induced right ventricular dysfunction". Experimental and Therapeutic Medicine 32.6 (2026): 347.
Chicago
Zhang, H., Zhao, Q., Sun, J., Deng, Y., Xing, G., Duan, W., Li, G., Wang, Q."Compensatory activation of PI3K/AKT signaling and apoptosis-associated protein expression in pulmonary hypertension‑induced right ventricular dysfunction". Experimental and Therapeutic Medicine 32, no. 6 (2026): 347. https://doi.org/10.3892/etm.2026.13342
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