Korean J Physiol Pharmacol.  2023 Jul;27(4):345-356. 10.4196/kjpp.2023.27.4.345.

Hydrogen sulfide ameliorates abdominal aorta coarctationinduced myocardial fibrosis by inhibiting pyroptosis through regulating eukaryotic translation initiation factor 2αα phosphorylation and activating PI3K/AKT1 pathway

• Li Y
• Wu Z
• Hu J
• Liu G
• Hu H
• Ouyang F
• Yang J

• KMID: 2544136
• DOI: http://doi.org/10.4196/kjpp.2023.27.4.345

Abstract

This study aimed to assess the effects of exogenous hydrogen sulfide (H₂S) on abdominal aorta coarctation (AAC) induced myocardial fibrosis (MF) and autophagy in rats. Forty-four Sprague–Dawley rats were randomly divided into control group, AAC group, AAC + H₂S group, and H₂S control group. After a model of rats with AAC was built surgically, AAC + H₂S group and H₂S group were injected intraperitoneally with H₂S (100 μmol/kg) daily. The rats in the control group and the AAC group were injected with the same amount of PBS. We observed that H₂S can improve left ventricular function and the deposition of myocardial collagen fibers, inhibit pyroptosis, down-regulate the expression of P-eif2α in myocardial tissue, and inhibit cell autophagy by activating the phosphatidylinositol 3-kinase (PI3K)/AKT1 signaling pathway (p < 0.05). In addition, angiotensin II (1 μM) H9c2 cardiomyocytes were injured in vitro experiments, and it was also observed that pyroptosis was inhibited after H₂S (400 μmol/kg) intervention, the expression of P-eif2α in cardiomyocytes was significantly down-regulated, and the PI3K/AKT1 signaling pathway was activated at the same time. Therefore, increasing the expression of P-eif2α reverses the activation of the PI3K/AKT1 signaling pathway by H₂S. In conclusion, these findings suggest that exogenous H₂S can ameliorate MF in rats with AAC by inhibiting pyroptosis, and the mechanism may be associated with inhibiting the phosphorylation of eif2α and activating the PI3K/AKT1 signaling pathway to inhibit excessive cell autophagy.

Keyword

Fibrosis; Hydrogen sulfide; Phosphatidylinositol 3-Kinase; Phosphorylation; Pyroptosis

Figure

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  Fig. 1 The contents hydrogen sulfide (H2S) of myocardial in abdominal aorta coarctation (AAC)-induced myocardial fibrosis treated with control, AAC, AAC + H2S, and H2S. Values are presented as mean ± SD. *p < 0.05 vs. control, #p < 0.05 vs. AAC.

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  Fig. 2 Upregulation of cardiac production of myocardial collagen fibers in abdominal aorta coarctation (AAC)-injected rat in vivo, this effect can be antagonized by hydrogen sulfide (H2S). Representative images of Masson with myocardial collagen fibers in the heart tissues of AAC-injected rat models (Blue represents collagen fibers). Images were acquired at 10 × 40 magnification. (A) Control group. (B) AAC group. (C) AAC + H2S group. (D) H2S group.

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  Fig. 3 Western blotting analysis the expression of (A) MMP1, (B) MMP13, (C) TIMP1 in abdominal aorta coarctation (AAC)-induced myocardial fibrosis treated with control, AAC, AAC + H2S, and H2S. Values are presented as mean ± SD. H2S, hydrogen sulfide. *p < 0.05 vs. control, #p < 0.05 vs. AAC, ##p < 0.01 vs. AAC.

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  Fig. 4 Western blotting analysis the expression of (A) Caspase1, (B) Caspase3 in abdominal aorta coarctation (AAC)-induced myocardial fibrosis treated with control, AAC, AAC + H2S, and H2S. Values are presented as mean ± SD. H2S, hydrogen sulfide. *p < 0.05 vs. control, **p<0.01 vs. control, ##p<0.01 vs. AAC.

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  Fig. 5 Western blotting analysis the expression of (A) ICAM-1, (B) IL6, IL1β, IL18 in abdominal aorta coarctation (AAC)-induced myocardial fibrosis treated with control, AAC, AAC + H2S, and H2S. Values are presented as mean ± SD. H2S, hydrogen sulfide. *p < 0.05 vs. control, ***p < 0.001 vs. control, #p < 0.05 vs. AAC, ##p < 0.01 vs. AAC, ###p < 0.001.

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  Fig. 6 Hydrogen sulfide (H2S) inhibitions abdominal aorta coarctation (AAC)-induced myocardial fibrosis mitophagy. Representative images of transmission electron microscopy (black arrows indicate mitophagy). Transmission electron micrographs at bars of 0.5 µm. (A) Control group. (B) AAC group. (C) AAC + H2S group. (D) H2S group.

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  Fig. 7 Western blotting analysis the expression of (A) Atg3, (B) Atg16L1, (C) Beclin1, (D) LC3A/B in abdominal aorta coarctation (AAC)-induced myocardial fibrosis treated with control, AAC, AAC + H2S, and H2S. Values are presented as mean ± SD. H2S, hydrogen sulfide. *p < 0.05 vs. control, #p < 0.05 vs. AAC.

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  Fig. 8 Western blotting analysis the expression of (A) P-eif2α, (B) P-PI3K, (C) P-AKT1 in abdominal aorta coarctation (AAC)-induced myocardial fibrosis treated with control, AAC, AAC + H2S, and H2S. Values are presented as mean ± SD. H2S, hydrogen sulfide. *p < 0.05 vs. control, #p < 0.05 vs. AAC, ##p < 0.01 vs. AAC.

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  Fig. 9 Western blotting analysis the expression of CTH in H9c2 cells treated with control, Ang II, Ang II + H2S, and PAG. Values are presented as mean ± SD. CTH, γ-Cystathionase; Ang II, angiotensin II; H2S, hydrogen sulfide; PAG, DL-Propargylglycine. **p < 0.01 vs. control, #p < 0.05 vs. Ang II, $p < 0.05 vs. Ang II + H2S.

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  Fig. 10 Western blotting analysis the expression of (A) P-eif2α, (B) P-PI3K, P-AKT1, (C) Caspase1, Caspase3 in H9c2 cells treated with control, Ang II, Ang II + H2S, and H2S. Values are presented as mean ± SD. Ang II, angiotensin II; H2S, hydrogen sulfide. *p < 0.05 vs. control, ***p < 0.001 vs. control, #p < 0.05 vs. Ang II, ##p < 0.01 vs. Ang II, ###p < 0.001 vs. Ang II.

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  Fig. 11 Western blotting analysis the expression of (A) P-eif2α, (B) P-PI3K, P-AKT1, (C) Caspase1, Caspase3 in H9c2 cells treated with control, Ang II, Ang II + H2S, Ang II + H2S + BTdCPU, and H2S. Values are presented as mean ± SD. Ang II, angiotensin II; H2S, hydrogen sulfide; BTdCPU, eif2α phosphorylation activation. *p < 0.05 vs. control, #p < 0.05 vs. Ang II, ##p < 0.01 vs. Ang II, $p < 0.05 vs. Ang II + H2S, $$p < 0.05 vs. Ang II + H2S.

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