丹皮酚调控p38MAPK-Nrf2信号通路减轻内皮细胞焦亡影响大鼠皮瓣缺血再灌注损伤机制OA
Paeonol alleviates rat skin flap ischemia-reperfusion injury by reducing endothelial cell pyroptosis via regulating p38MAPK-Nrf2 signaling pathway
[目的]探讨丹皮酚改善大鼠皮瓣缺血再灌注损伤(I/R)的机制.[方法]60只雄性Sprague-Dawley大鼠随机分为4组:假手术(Sham)组、I/R组、丹皮酚组、丹皮酚+ML385组,每组15只.除Sham组外,其他组建立 I/R损伤皮瓣模型.丹皮酚组、丹皮酚+ML385组给予50 mg/kg丹皮酚治疗.丹皮酚+ML385组腹腔注射30 mg/kg ML385.7d后,分析大鼠皮瓣存活面积和皮瓣中的血流量.通过免疫荧光染色分析皮瓣中肌动蛋白α2(ACTA2)、CD31、消皮素D蛋白N端结构域(GSDMD-N)表达情况,二氢乙锭染色分析活性氧(ROS)水平,蛋白免疫印迹分析P38丝裂原激活蛋白激酶(p38MAPK)-核因子E2相关因子2(Nrf2)信号.[结果]与Sham组相比,I/R组大鼠的皮瓣存活面积、皮瓣血流信号强度、CD31 和 ACTA2 共定位阳性血管面积、血管内皮生长因子(VEGF)、基质金属蛋白酶 9(MMP9)蛋白表达降低(P<0.05),CD31和GSDMD-N共定位阳性细胞数增加(P<0.05).与I/R组相比,丹皮酚组大鼠的皮瓣存活面积、皮瓣血流信号强度、CD31和ACTA2共定位阳性血管面积以及VEGF、MMP9血管生成相关蛋白表达增加(P<0.05),CD31和GSDMD-N共定位阳性细胞数明低(P<0.05).丹皮酚组大鼠皮瓣ROS的相对荧光强度、丙二醛(MDA)水平较I/R组降低(P<0.05),还原型谷胱甘肽(GSH)水平较I/R组增加(P<0.05).与丹皮酚组相比,丹皮酚+ML385组大鼠的皮瓣中Nrf2蛋白表达减少(P<0.05),p-P38蛋白表达增加(P<0.05).此外,ML385加入逆转了丹皮酚对皮瓣I/R大鼠模型皮瓣存活面积、皮瓣血流信号强度、血管生成、细胞焦亡、氧化应激的改善作用.[结论]丹皮酚通过增强皮瓣I/R模型大鼠抗氧化防御系统,保护真皮的血管内皮细胞免受氧化应激损伤,减轻了细胞焦亡,其作用机制可能与调控p38MAPK-Nrf2信号通路有关.
[Objective]To explore the mechanism of paeonol in improving skin flap ischemia-reperfusion injury(I/R)in rats.[Methods]Sixty male Sprague-Dawley rats were randomly divided into four groups:sham group,I/R group,paeonol group,and paeonol+ML385 group,with 15 rats in each group.Except for the sham group,the other groups were used to establish flap I/R injury models.The paeonol group and paeonol+ML385 group were treated with 50 mg/kg paeonol.The paeonol+ML385 group was intraperitoneally injected with 30 mg/kg ML385.After 7 days,the survival area of the rat flaps and blood flow in the flaps were analyzed.The expressions of actin alpha cardiac muscle 2(ACTA2),CD31,and the N-terminal domain of gasdermin D(GSDMD-N)in the flaps were analyzed by immunofluorescence staining.Reactive oxygen species(ROS)levels were analyzed by dihydroethidium staining.The p38 mitogen-activated protein kinase(p38MAPK)-nuclear factor E2-related factor 2(Nrf2)signaling pathway was analyzed by Western blot.[Results]Compared with the sham group,the I/R group showed significantly decreased flap survival area,flap blood flow signal intensity,area of CD31 and ACTA2 co-localized positive vessels,and protein expressions of vascular endothelial growth factor(VEGF)and matrix metalloproteinase 9(MMP9)(P<0.05),while the number of CD31 and GSDMD-N co-localized positive cells was significantly increased(P<0.05).Compared with the I/R group,the paeonol group showed significantly increased flap survival area,flap blood flow signal intensity,area of CD31 and ACTA2 co-localized positive vessels,and protein expressions of VEGF and MMP9(P<0.05),while the number of CD31 and GSDMD-N co-localized positive cells was significantly decreased(P<0.05).The relative fluorescence intensity of ROS and malondialdehyde(MDA)level in the flap of the paeonol group were significantly lower than those in the I/R group(P<0.05),while the glutathione(GSH)level was significantly higher(P<0.05).Compared with the paeonol group,the paeonol+ML385 group showed significantly decreased Nrf2 protein expression(P<0.05)and significantly increased p-p38 protein expression(P<0.05)in the flaps.Furthermore,the addition of ML385 reversed the beneficial effects of paeonol on flap survival area,blood flow signal intensity,angiogenesis,pyroptosis,and oxidative stress in the rat flap I/R model.[Conclusion]Paeonol protects dermal vascular endothelial cells from oxidative stress injury and reduces pyroptosis by enhancing the antioxidant defense system in rats with flap I/R injury.Its mechanism may be related to the regulation of the p38MAPK-Nrf2 signaling pathway.
陈宇虹;周青;唐庆妮;陈姣凤
湖南中医药大学第一附属医院手外创伤科,长沙 410000湖南中医药大学第一附属医院手外创伤科,长沙 410000湖南中医药大学第一附属医院手外创伤科,长沙 410000湖南中医药大学第一附属医院手外创伤科,长沙 410000
医药卫生
丹皮酚P38丝裂原激活蛋白激酶大鼠皮瓣缺血再灌注损伤氧化应激
paeonolp38 mitogen-activated protein kinaseratskin flapischemia-reperfusion injuryoxidative stress
《天津中医药大学学报》 2026 (4)
428-435,8
湖南省卫生健康委员会科研项目(20241317).
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