督脉隔药灸联合水凝胶多功能复合敷料对压力性损伤大鼠Nrf2/HO-1/NQO1信号通路的影响

Effects of Du meridian moxibustion on medicines combined with multifunctional composite hydrogel dressing on Nrf2/HO-1/NQO1 signaling pathway in rats with pressure injury

目的观察督脉隔药灸联合水凝胶多功能复合敷料对压力性损伤大鼠创面修复及核因子E2相关因子2(nuclear factor-erythroid 2 related factor 2,Nrf2)/血红素氧合酶1(heme oxygenase-1,HO-1)/NADPH醌氧化还原酶1(NADPH quinone oxidoreductase 1,NQO1)信号通路的影响。方法选取50只SPF级SD大鼠,随机分为假手术组、模型组、水凝胶敷料组、督脉隔药灸组、联合干预组,每组10只。除假手术组外,其余4组借助压力装置构建压力性损伤大鼠模型。分组干预并记录各组大鼠的压力性损伤修复情况及创面组织病理变化;借助ELISA检测各组大鼠的血清炎性因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素(interleukin,IL)-1β、IL-6以及氧化应激因子丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)水平;Western blot与RT-PCR检测各组大鼠创面肉芽组织Nrf2、HO-1、NQO1、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)蛋白及mRNA的表达水平。结果HE染色结果显示:假手术组皮肤结构未被破坏;模型组创面肉芽组织可见明显的细胞坏死、变性及炎性细胞浸润;干预各组可见创面修复,表现为散在炎性细胞浸润,“气球样”坏死变性明显减少。Masson染色结果显示:假手术组创面皮肤组织胶原纤维较为整齐;模型组创面肉芽组织可见大面积的胶原纤维沉积、紊乱;干预各组可见不同程度的创面修复。与假手术组对比,模型组大鼠血清TNF-α、IL-1β、IL-6、MDA水平和创面肉芽组织Nrf2、HO-1、NQO1、MMP-9蛋白及mRNA的表达水平均升高(P<0.05),而创面修复率、血清SOD、GSH-Px水平均下降(P<0.05);与模型组比较,水凝胶敷料组、督脉隔药灸组、联合干预组的血清TNF-α、IL-1β、IL-6、MDA水平均下降(P<0.05),而创面修复率、血清SOD、GSH-Px水平和创面肉芽组织Nrf2、HO-1、NQO1、MMP-9蛋白及m RNA的表达水平均升高(P<0.05),且联合干预组较其他干预组更优(P<0.05)。结论督脉隔药灸可有效促进压力性损伤的创面修复,与水凝胶多功能复合敷料联合使用效果更佳,其机制可能与督脉隔药灸抑制创面炎性反应及氧化应激,并激活Nrf2/HO-1/NQO1信号通路相关。

Objective To observe the effects of Du meridian moxibustion on medicines combined with multifunctional composite hydrogel dressing on wound repair and nuclear factor-erythroid 2 related factor 2(Nrf2)/heme oxygenase-1(HO-1)/NADPH quinone oxidoreductase 1(NQO1)signaling pathway in rats with pressure injury.Methods A total of 50 SPF SD rats were randomly divided into sham-operated group,model group,hydrogel dressing group,Du meridian moxibustion on medicines group,and combined intervention group,with ten rats in each group.Except for the sham-operated group,pressure injury rat models were established in the remaining four groups using pressure devices.The intervention was given to each group respectively,and the repair of pressure injury and pathological changes of wound tissue of rats in each group were recorded.ELISA was used to determine the levels of serum inflammatory factors including tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,and IL-6,as well as oxidative stress factors including malondialdehyde(MDA),superoxide dismutase(SOD),and glutathione peroxidase(GSH-Px)in each group.Western blot and RT-PCR were used to check the protein and mRNA expression levels of Nrf2,HO-1,NQO1,and matrix metalloproteinase-9(MMP-9)in wound granulation tissue of rats in each group.Results The HE staining showed that the skin structure of sham-operated group was not damaged;significant cell necrosis,degeneration,and inflammatory cell infiltration of wound granulation tissue were observed in the model group;wound was repaired in all intervention groups,manifested as scattered infiltration of inflammatory cells and a significant reduction in"balloon like"necrosis and degeneration.The Masson staining showed that the collagen fibers in the wound skin tissue of the sham-operated group were relatively neat;large areas of collagen fiber deposition and disorder in wound granulation tissue were observed in the model group;all intervention groups showed varying degrees of wound repair.Compared with sham-operated group,the serum levels of TNF-α,IL-1β,IL-6,and MDA,as well as the protein and mRNA expression levels of Nrf2,HO-1,NQO1,MMP-9 in wound granulation tissue of the model group were higher(P<0.05),while the wound repair rate,serum levels of SOD and GSH-Px were lower(P<0.05).Compared with model group,the serum levels of TNF-α,IL-1β,IL-6,and MDA in hydrogel dressing group,Du meridian moxibustion on medicines group,and combined intervention group decreased(P<0.05),while the wound repair rate,the serum levels of SOD and GSH-Px,and the protein and mRNA expression levels of Nrf2,HO-1,NQO1,and MMP-9 in wound granulation tissue increased(P<0.05),and the combined intervention group showed better results than other intervention groups(P<0.05).Conclusion Du meridian moxibustion on medicines can effectively promote wound repair in pressure injury and the effect is better when combined with multifunctional composite hydrogel dressing.Its mechanism may be related to the inhibition of wound inflammatory reaction and oxidative stress,as well as the activation of Nrf2/HO-1/NQO1 signaling pathway by Du meridian moxibustion on medicines.