Yu Pan, Hong Nan, Wang Genwang, Chen Shuqiang, Zhao Zhipeng
Department of Burn and Plastic Surgery, Jinling Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
Department of Scientific Research and Training, Jinling Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
Cytojournal. 2025 Apr 25;22:46. doi: 10.25259/Cytojournal_253_2024. eCollection 2025.
Burns refers to a severe form of trauma that often leads to localized and systemic inflammatory responses, oxidative stress, and immune dysfunction. Patients with severe burns are highly susceptible to the development of postburn sepsis, a condition influenced by multiple factors, such as bacterial infection of the burn wound, alterations in immune status, and excessive release of inflammatory mediators. This study aimed to investigate the mechanisms by which hydrogen gas treatment exerts its effects on postburn sepsis, with a focus on its influence on inflammatory responses, oxidative stress, and wound healing.
This work employed assays with Sprague-Dawley (SD) rat skin fibroblasts (RSFs) to assess the effects of burn serum and hydrogen gas on cell proliferation through methylthiazolyldiphenyltetrazolium bromide assays and on apoptosis through flow cytometry with Annexin V-fluorescein isothiocyanate/propidium iodide staining. In addition, an enzyme-linked immunosorbent assay was performed to quantify inflammatory cytokines and oxidative stress markers in fibroblasts treated with burn serum. Western blotting (WB) analysis was conducted to investigate signaling pathway modulation. The severe burn sepsis models of SD rats were segregated into three experimental groups: a healthy normal control group, a burn sepsis control group, and a burn sepsis + hydrogen gas (2%) treatment group. Wound healing was monitored, with wound contraction rates recorded and histological assessments conducted using hematoxylin and eosin and Masson's trichrome staining to evaluate tissue repair and collagen deposition.
assays showed that burn serum reduced fibroblast proliferation and increased apoptosis ( < 0.01), which hydrogen gas mitigated by rescuing cell viability and reducing apoptosis ( < 0.01). Enzyme-linked immunosorbent assay revealed burn serum-induced increases in the levels of inflammatory cytokines and oxidative stress markers, with decreases in antioxidant enzymes ( < 0.01), which hydrogen gas reversed ( < 0.05). WB analysis suggested hydrogen gas's anti-inflammatory and proliferative effects by modulating signaling pathways ( < 0.01). , hydrogen gas treatment considerably improved wound healing, with accelerated contraction and enhanced collagen deposition. Plasma and skin tissue analyses indicated systemic and local anti-inflammatory and antioxidant effects from hydrogen gas.
Hydrogen gas treatment demonstrates potential therapeutic efficacy in the management of postburn sepsis by modulating inflammatory responses, reducing oxidative stress, and promoting wound healing. These findings provide scientific evidence supporting hydrogen gas as an adjunctive treatment strategy for postburn sepsis.
烧伤是一种严重的创伤形式，常导致局部和全身的炎症反应、氧化应激及免疫功能障碍。严重烧伤患者极易发生烧伤后脓毒症，这一病症受多种因素影响，如烧伤创面的细菌感染、免疫状态改变及炎症介质的过度释放。本研究旨在探究氢气治疗对烧伤后脓毒症发挥作用的机制，重点关注其对炎症反应、氧化应激及伤口愈合的影响。
本研究采用Sprague-Dawley（SD）大鼠皮肤成纤维细胞（RSF）进行实验，通过甲基噻唑基二苯基四氮唑溴盐实验评估烧伤血清和氢气对细胞增殖的影响，并通过膜联蛋白V-异硫氰酸荧光素/碘化丙啶染色的流式细胞术评估对细胞凋亡的影响。此外，进行酶联免疫吸附测定以量化用烧伤血清处理的成纤维细胞中的炎性细胞因子和氧化应激标志物。进行蛋白质免疫印迹（WB）分析以研究信号通路调节。将SD大鼠的严重烧伤脓毒症模型分为三个实验组：健康正常对照组、烧伤脓毒症对照组和烧伤脓毒症+氢气（2%）治疗组。监测伤口愈合情况，记录伤口收缩率，并使用苏木精和伊红以及Masson三色染色进行组织学评估，以评估组织修复和胶原沉积。
实验表明，烧伤血清降低了成纤维细胞的增殖并增加了细胞凋亡（P<0.01），而氢气通过挽救细胞活力和减少细胞凋亡减轻了这种情况（P<0.01）。酶联免疫吸附测定显示烧伤血清诱导炎性细胞因子和氧化应激标志物水平升高，抗氧化酶水平降低（P<0.01），而氢气使其逆转（P<0.05）。WB分析表明氢气通过调节信号通路发挥抗炎和增殖作用（P<0.01）。此外，氢气治疗显著改善了伤口愈合，加速了收缩并增强了胶原沉积。血浆和皮肤组织分析表明氢气具有全身和局部抗炎及抗氧化作用。
氢气治疗通过调节炎症反应、降低氧化应激和促进伤口愈合，在烧伤后脓毒症的治疗中显示出潜在的治疗效果。这些发现为氢气作为烧伤后脓毒症的辅助治疗策略提供了科学证据。
