Hydrogen Therapy Protects Muscle During Limb Reattachment Surgery
- Authors
- Zi-Hao Jiang, Jun-Sheng Wang, Jin-Ling Wang, Jiang-Fan Zheng, Xiao-Ling Li, Zhi-Cheng Yang, Meng-Qiu Xu, Yong-Li Zhang, Yu Wang
- Journal
- Heliyon
- Year
- 2024
- DOI
- 10.1016/j.heliyon.2024.e37018
- Study Type
- Rat
- Outcome
- Positive
- Peer Reviewed
- Yes
- Country
- China
- Health Condition
- Ischemia-Reperfusion Injury
- Body System
- Musculoskeletal
TL;DR
Pre-treating with hydrogen-rich heparin sodium helps protect muscles from damage after limb reattachment by boosting the body's antioxidant defenses.
Key Finding
Hydrogen-rich heparin activated a cellular protection pathway (NRF2/HO-1) that reduced oxidative stress and cell death in muscle tissue after simulated limb replantation in rats.
Summary
This rat study tested whether hydrogen-rich heparin (a blood thinner mixed with hydrogen) could reduce damage to leg muscles during limb replantation surgery. When a limb is reattached, blood flow is restored after being cut off, which causes oxidative stress (harmful chemical reactions) and cell death. Researchers found that hydrogen-rich heparin activated a protective cellular pathway (NRF2/HO-1) that reduced oxidative damage and prevented cells from dying, compared to regular heparin alone.
Practical Takeaway
This is early-stage research in rats only, so it cannot yet be applied to human surgery. The findings suggest that hydrogen-enriched solutions may help protect muscle tissue during limb replantation procedures, but human clinical trials would be needed to determine if this approach is safe and effective in actual surgical patients.
Abstract
Background: Ischaemia-reperfusion injury (IRI) is a critical complication post-limb replantation. The oxidative stress and cellular apoptosis due to IRI considerably hinder the healing process. This study aimed to investigate the modulatory effects of pre-perfusion with hydrogen-rich heparin sodium on the nuclear factor erythroid 2-related factor 2 (NRF2)/haeme oxygenase-1 (HO-1) pathway and its potential mechanisms in mitigating skeletal muscle IRI post-limb replantation. Methods: Forty healthy Sprague-Dawley rats (250-300 g) were classified into five groups (n = 8 each): normal control, IRI + heparin sodium pre-perfusion (heparin group), IRI + hydrogen-rich heparin sodium pre-perfusion (hydrogen-rich heparin group), IRI + hydrogen-rich heparin sodium pre-perfusion + NRF2 inhibitor (hydrogen-rich heparin + all-trans retinoic acid [ATRA] group), and IRI + heparin sodium pre-perfusion + NRF2 inhibitor (heparin + ATRA group). The activation of the NRF2/HO-1 pathway in skeletal muscle IRI was evaluated based on HO-1 expression using western blotting and immunofluorescence. Furthermore, haematoxylin and eosin staining and transmission electron microscopy were employed to determine the histopathological characteristics. Additionally, superoxide dismutase and malondialdehyde levels in skeletal muscle tissue were measured to assess antioxidant capacity and the degree of oxidative stress damage. Tissue hypoxia was assessed based on hypoxia-inducible factor 1-alpha expression, whereas apoptosis markers BCL-2-associated X protein (BAX) and Caspase-3 in skeletal muscle tissues were analysed using western blotting with terminal deoxynucleotidyl transferase dUTP nick end labelling staining to quantify cell apoptosis. Results: Compared with the control group, the heparin group exhibited significant pathological changes, including inflammatory infiltration and cellular hypertrophy, with increased apoptosis and oxidative stress. Notably, NRF2 suppression aggravated these effects. However, hydrogen-rich heparin sodium prominently activated the NRF2/HO-1 pathway, enhancing antioxidant defence and reducing BAX/Caspase-3-mediated apoptosis, thereby mitigating IRI-induced damage. The use of an NRF2 inhibitor to inhibit NRF2 excitation by hydrogen-rich heparin sodium notably weakened NRF2 activation and the antioxidant response, resulting in a substantial increase in cellular apoptosis. Conclusion: Pre-perfusion with hydrogen-rich heparin sodium markedly diminishes the BAX/Caspase-3-mediated apoptotic pathway in skeletal muscle tissues with IRI through the excitation of the NRF2/HO-1 pathway.