氢气通过抑制氧化应激促进创面愈合

Hydrogen promotes wound healing by inhibiting oxidative stress

目的:探讨氢气对创面愈合产生的影响及其分子机制。方法:将36只昆明小鼠采用随机数字表法分为对照组、腹腔注射生理盐水组、腹腔注射富氢生理盐水组,每组12只。记录创面愈合时间并在第3、7天采集创面进行组织学观察。采用H 2O 2对原代分离的人成纤维细胞进行氧化刺激,检测细胞活力及细胞活性氧(ROS)水平,通过免疫荧光实验检测核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)的表达水平,并通过t检验评估组间差异。结果:腹腔注射富氢生理盐水组的创面愈合时间较对照组和腹腔注射生理盐水组更短[(11.17±0.68)d比(13.67±0.94)d,t=4.792,P<0.01;(11.17±0.68)d比(13.33±0.47)d,t=5.814,P<0.01]。在H 2O 2介导的氧化应激下,经富氢培养基处理的人成纤维细胞活力较标准培养基高[(58.64±15.19)%比(29.13±4.07)%,t=3.250,P<0.05]。加入Nrf2特异性抑制剂后,经富氢培养基处理的人成纤维细胞Nrf2荧光表达降低(0.44±0.06比1.64±0.33,t=8.632,P<0.01),HO-1荧光表达降低(0.42±0.06比1.36±0.14,t=15.130,P<0.01),细胞活力降低[(112.81±20.05)%比(156.41±10.78)%,t=4.692,P<0.01]。结论:氢通过激活Nrf2/HO-1通路缓解氧化应激反应从而促进创面愈合。

Objective To investigate the effects of hydrogen on wound healing and its underlying molecular mechanisms.Methods Totally,36 Kunming mice purchased from the experimental animal center were randomly divided into the three groups by the random number table:control group,positive control group given the intraperitoneal injection of saline group,and hydrogen group given the intraperitoneal injection of hydrogen-rich saline.The wound healing time was recorded,and samples were collected on the 3rd and 7th day for histopathological examination.The primary isolated fibroblasts were used as model cells and H2O2 treatment was used as oxidative stimulation.The cell viability,cellular reactive oxygen species,and expression of nuclear factor erythroid-2 related factor 2(Nrf2)and heme oxygenase 1(HO-1)were detected.Differences between groups were evaluated by t test.Results The wound healing time in hydrogen group was significantly shorter than in control group and positive control group[(11.17±0.68)d vs.(13.67±0.94)d,t=4.792,P<0.01;(11.17±0.68)d vs.(13.33±0.47)d,t=5.814,P<0.01].The hydrogen-rich medium improved the cell viability under the oxidative stimulation of H2O2[(58.64±15.19)%vs.(29.13±4.07)%,t=3.250,P<0.05].The application of Nrf2-inhibitor in hydrogen-rich medium reduced the expression of Nrf2(0.44±0.06 vs.1.64±0.33,t=8.632,P<0.01),its downstream protein HO-1(0.42±0.06 vs.1.36±0.14,t=15.130,P<0.01)and cell viability[(112.81±20.05)%vs.(156.41±10.78)%,t=4.692,P<0.01]in fibroblasts.Conclusion Hydrogen promotes wound healing by relieving oxidative stress via the Nrf2/HO-1 pathway.