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Prenatal lipopolysaccharide exposure programs cardiac fibrosis via dysregulating of connexin 43 in offspring rats

  • Authors:
    • Yan Wen
    • Yao Yuan
    • Haigang Zhang
    • Ya Liu
  • View Affiliations / Copyright

    Affiliations: Department of Pharmacology, College of Pharmacy, Army Medical University, Chongqing 400038, P.R. China
    Copyright: © Wen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 120
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    Published online on: February 24, 2026
       https://doi.org/10.3892/mmr.2026.13830
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Abstract

The present study investigated the role of connexin 43 (Cx43) in mediating prenatal inflammation‑induced cardiac fibrosis in offspring, specifically exploring its dynamic regulation with autophagy and DNA methylation pathways. Pregnant Sprague‑Dawley rats received intraperitoneal injections of saline (control) or lipopolysaccharide (LPS, 0.79 mg/kg) on gestational days 8, 10 and 12. Offspring were sacrificed at 8 and 16 weeks postpartum. Myocardial tissues were subjected to histopathological examination and molecular analysis. Prenatal LPS exposure consistently induced significant cardiac fibrosis in the offspring. Reverse transcription‑quantitative PCR revealed that mRNA levels of Cx43, LC3 and DNA methyltransferase 1 (DNMT1) were markedly reduced at 8 weeks; however, they were elevated above control levels at 16 weeks. Western blotting revealed persistent suppression of Cx43 protein expression at both ages, whereas the LC3‑II/I ratio and DNMT1 protein levels paralleled the biphasic mRNA trends. In vitro experiments using neonatal rat cardiac fibroblasts treated with LPS (10 µg/ml, 24 h) confirmed Cx43 and LC3 downregulation and DNMT1 upregulation. Targeted pharmacological interventions were used to clarify these regulatory relationships. Cotreatment with the Cx43 gap junction inhibitor carbenoxolone (400 µM) and LPS further suppressed Cx43, LC3 and DNMT1 expression. However, cotreatment with the Cx43 agonist all‑trans retinoic acid (10 µM) attenuated LPS‑induced DNMT1 upregulation and LC3‑II/I ratio suppression. These findings demonstrate that the functional state of Cx43 critically links fetal inflammatory insults to postnatal cardiac fibrogenesis by dynamically regulating interconnected autophagy and DNA methylation, establishing Cx43 as an upstream regulatory node in this pathogenic network.

View Figures

Figure 1

BW of offspring rats from 1-day to
16-week-old (A). Heart damages in offspring at the age of 8 and 16
weeks, including the ratios (B) LVW/BW, (C) HW/BW and (D) NT-proBNP
level in serum. Concentration of Ang II in (E) left ventricle and
(F) serum. Data are presented as mean ± SD. n=10 in each group
(A-C) and n=7 in each group (D-F). *P<0.05, **P<0.01 vs Con.
BW, body weight; HW, heart weight; LVW, left ventricular weight;
NT-proBNP, N-terminal pro-brain natriuretic peptide; Ang II,
angiotensin II; Con, control; LPS, lipopolysaccharide.

Figure 2

Histopathological observation of LV
in 8- and 16-week-old offspring rats. (A) Hematoxylin and eosin
staining. (B) Masson trichrome staining and (C) CVF. Data are
presented as mean ± SD. n=6 in each group. LV, left ventricular;
CVF, collagen volume fraction; Con, control; LPS,
lipopolysaccharide.

Figure 3

Immunolabeling of α-smooth muscle
actin in the heart. Con, control; LPS, lipopolysaccharide.

Figure 4

mRNA expression of Cx43, LC3, DNMT1,
DNMT3A and DNMT3B in the LV from offspring rats at the age of (A) 8
and (B) 16 weeks. Data are presented as mean ± SD. n=6 in each
group. Cx43, connexin 43; DNMT, DNA methyltransferase; LV, left
ventricular; Con, control; LPS, lipopolysaccharide.

Figure 5

Protein expression of Cx43, LC3II/I,
DNMT1, DNMT3A and DNMT3B in LV from offspring rat at (A) 8 and (B)
16 weeks. n=6 in each group. Cx43, connexin 43; DNMT, DNA
methyltransferase; LV, left ventricular; Con, control; LPS,
lipopolysaccharide.

Figure 6

Effect of Cx43 on the expression of
LC3 and DNMT1 in vitro. (A) mRNA and (B) protein expression
of Cx43, LC3 and DNMT1 in rat primary cardiac fibroblasts. Data are
presented as mean ± SD. n=3/group. Control group, rat primary
cardiac fibroblasts treated with culture media; LPS group, rat
primary cardiac fibroblasts treated with culture media containing
10 µg/ml LPS; LPS + CBX group, rat primary cardiac fibroblasts
treated with culture media containing 10 µg/ml LPS and 400 µM CBX;
LPS + ATRA group, rat primary cardiac fibroblasts treated with
culture media containing 10 µg/ml LPS and 10 µM all-trans retinoic
acid. Cx43, connexin 43; DNMT, DNA methyltransferase; LPS,
lipopolysaccharide; CBX, carbenoxolone; ATRA, all-trans retinoic
acid; Con, control; LPS, lipopolysaccharide.
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Copy and paste a formatted citation
Spandidos Publications style
Wen Y, Yuan Y, Zhang H and Liu Y: <p>Prenatal lipopolysaccharide exposure programs cardiac fibrosis via dysregulating of connexin 43 in offspring rats</p>. Mol Med Rep 33: 120, 2026.
APA
Wen, Y., Yuan, Y., Zhang, H., & Liu, Y. (2026). <p>Prenatal lipopolysaccharide exposure programs cardiac fibrosis via dysregulating of connexin 43 in offspring rats</p>. Molecular Medicine Reports, 33, 120. https://doi.org/10.3892/mmr.2026.13830
MLA
Wen, Y., Yuan, Y., Zhang, H., Liu, Y."<p>Prenatal lipopolysaccharide exposure programs cardiac fibrosis via dysregulating of connexin 43 in offspring rats</p>". Molecular Medicine Reports 33.4 (2026): 120.
Chicago
Wen, Y., Yuan, Y., Zhang, H., Liu, Y."<p>Prenatal lipopolysaccharide exposure programs cardiac fibrosis via dysregulating of connexin 43 in offspring rats</p>". Molecular Medicine Reports 33, no. 4 (2026): 120. https://doi.org/10.3892/mmr.2026.13830
Copy and paste a formatted citation
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Spandidos Publications style
Wen Y, Yuan Y, Zhang H and Liu Y: <p>Prenatal lipopolysaccharide exposure programs cardiac fibrosis via dysregulating of connexin 43 in offspring rats</p>. Mol Med Rep 33: 120, 2026.
APA
Wen, Y., Yuan, Y., Zhang, H., & Liu, Y. (2026). <p>Prenatal lipopolysaccharide exposure programs cardiac fibrosis via dysregulating of connexin 43 in offspring rats</p>. Molecular Medicine Reports, 33, 120. https://doi.org/10.3892/mmr.2026.13830
MLA
Wen, Y., Yuan, Y., Zhang, H., Liu, Y."<p>Prenatal lipopolysaccharide exposure programs cardiac fibrosis via dysregulating of connexin 43 in offspring rats</p>". Molecular Medicine Reports 33.4 (2026): 120.
Chicago
Wen, Y., Yuan, Y., Zhang, H., Liu, Y."<p>Prenatal lipopolysaccharide exposure programs cardiac fibrosis via dysregulating of connexin 43 in offspring rats</p>". Molecular Medicine Reports 33, no. 4 (2026): 120. https://doi.org/10.3892/mmr.2026.13830
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