基于NLRP3/Caspase-1信号通路探讨溃疡油对慢性创面的抗炎促愈作用

Exploration on the anti-inflammatory and healing effects of ulcer oil on chronic wounds based on NLRP3/Caspase-1 signaling pathway

目的:观察溃疡油对慢性皮肤溃疡新西兰大白兔的创面愈合作用,以及对创面组织核苷酸结合寡聚结构域样受体蛋白3(NLRP3)、天冬氨酸特异性半胱氨酸蛋白酶1(Caspase-1)、核因子κB(NF-κB)表达的影响。方法:将60只新西兰大白兔随机分为空白组(15只)、造模组(45只)。采用“激素干预-皮肤缺损-细菌感染”复合因素法复制慢性皮肤溃疡模型,成功后随机分为模型对照组、乳酸依沙吖啶组、溃疡油组,每组15只,分别予0.9%氯化钠溶液、乳酸依沙吖啶、溃疡油干预21 d。分别在干预7、14、21 d时间点检测各组创面面积、创面组织增殖细胞核抗原(PCNA)、NLRP3、Caspase-1、NF-κB p65蛋白表达,以及血常规、肝功能、肾功能等安全性指标。结果:干预7 d时,与模型对照组和乳酸依沙吖啶组比较,溃疡油组创面面积显著减小(P<0.05);与空白组比较,模型对照组PCNA蛋白表达显著降低(P<0.01),NF-κB p65蛋白表达显著增加(P<0.01);与模型对照组和乳酸依沙吖啶组比较,溃疡油组PCNA蛋白表达显著增加(P<0.01);与模型对照组比较,乳酸依沙吖啶组和溃疡油组NF-κB p65蛋白表达均显著降低(P<0.01)。干预14 d时,与模型对照组和乳酸依沙吖啶组比较,溃疡油组创面面积显著减小(P<0.01),PCNA蛋白表达显著增加(P<0.05);与模型对照组比较,乳酸依沙吖啶组和溃疡油组NLRP3和NF-κB p65蛋白表达显著降低(P<0.01,P<0.05)。干预21 d时,与模型对照组比较,乳酸依沙吖啶组和溃疡油组创面面积均显著减小(P<0.01);与空白组比较,模型对照组PCNA蛋白表达显著降低(P<0.01),NF-κB p65蛋白表达显著增加(P<0.01);与模型对照组和乳酸依沙吖啶组比较,溃疡油组PCNA蛋白表达显著增加(P<0.01);与模型对照组比较,乳酸依沙吖啶组和溃疡油组NF-κB p65蛋白表达显著降低(P<0.05,P<0.01)。结论:溃疡油可能通过下调NLRP3/Caspase-1信号通路关键蛋白表达,尤其是NLRP3和NF-κB p65蛋白,以减轻创面炎症反应,同时促进创面组织细胞增殖,进而加速创面愈合,且干预过程中具有良好的安全性。

Objective:To observe the wound healing effect of ulcer oil on rabbits with chronic skin ulcers and the expression of nucleotide-binding oligodomain-like receptor protein 3(NLRP3),Caspase-1,and nuclear factor-kappa B(NFkB)in the wound tissue.Methods:A total of 60 New Zealand white rabbits were randomly divided into blank group(n=15)and molded group(n=45).The chronic skin ulcer model was replicated by"hormonal intervention-skin defect-bacterial infection'compound factor method.After success,it was randomly divided into model control group,ethacridine group and ulcer oil group.15 mice in each group were treated with 0.9%sodium chloride solution,ethacridine lactate and ulcer oil for 21 d.Wound area,tissue proliferating cell nuclear antigen(PCNA),NLRP3,Caspase-1,NF-kB p65 protein expression,and blood routine,liver function,kidney function and other safety indicators were detected at intervention 7,14 and 21 d time points.Results:At 7 days of intervention,compared with the model control group and the lactate ethacridine group,the ulcer oil group showed a significant reduction in wound area(P<0.05);Compared with the blank group,the model control group showed a significant decrease in PCNA protein expression(P<0.01)and a significant increase in NF-kB p65 protein expression(P<0.01);Compared with the model control group and the lactate ethacridine group,the expression of PCNA protein in the ulcer oil group was significantly increased(P<0.01);Compared with the model control group,the NF-kB p65 protein expression was significantly reduced in the lactate ethacridine group and ulcer oil group(P<0.01).At 14 days of intervention,compared with the model control group and the lactate ethacridine group,the ulcer oil group showed a significant decrease in wound area(P<0.01)and a significant increase in PCNA protein expression(P<0.05);Compared with the model control group,the expression of NLRP3 and NF-kB p65 proteins was significantly reduced in the lactate ethacridine group and ulcer oil group(P<0.01,P<0.05).At 21 days of intervention,compared with the model control group,both the lactate ethacridine group and the ulcer oil group showed a significant reduction in wound area(P<0.01);Compared with the blank group,the model control group showed a significant decrease in PCNA protein expression(P<0.01)and a significant increase in NF-kB p65 protein expression(P<0.01);Compared with the model control group and the lactate ethacridine group,the expression of PCNA protein in the ulcer oil group was significantly increased(P<0.01);Compared with the model control group,the NF-kB p65 protein expression was significantly reduced in the lactate ethacridine group and ulcer oil group(P<0.05,P<0.01).Conclusion:Ulcer oil may reduce wound inflammation by downregulating the expression of key proteins in the NLRP3/Caspase-1 signaling pathway,especially NLRP3 and NF-kB p65 proteins,while promoting wound tissue cell proliferation,accelerating wound healing,and having good safety during the intervention process.