猪脱细胞真皮基质对大鼠瘘管模型的修复作用及其对TGF-β1/NF-κB p65信号通路的影响

The effect of porcine acellular dermal matrix on fistula model repairing in rats and on TGF-β1/NF-κB p65 signaling pathway

目的:观察猪脱细胞真皮基质对大鼠瘘管模型创面的修复作用,以及对TGF-β1/NF-κB p65信号通路的影响,探讨其可能的修复机制。方法:40只健康SD大鼠适应性喂养1周后,随机分为假手术对照组、模型组、猪脱细胞真皮基质组(真皮组)、瘘管栓阳性对照组(瘘管栓组),每组10只。连续给予相应材料治疗2周后,采用ELISA法检测血清中白介素-1β(IL-1β)、白介素-6(IL-6)、血管内皮生长因子(VEGF)和前列腺素E2(PGE2)含量;取大鼠创面组织进行病理观察;Western blot法检测创面组织转化生长因子-β1(TGF-β1)、核转录因子-κB p65(NF-κB p65)及VEGF蛋白表达水平。结果:与模型组相比,治疗2周后真皮组和瘘管栓组大鼠的体质量明显增加(P<0.05),创面脓液明显减少。真皮组中血清IL-1β、IL-6和PGE2的含量比模型组低(P<0.05),VEGF含量比模型组高(P<0.05)。HE染色显示,与模型组相比,真皮组炎细胞浸润明显减少,胶原纤维明显增多并且更加致密。真皮组及瘘管栓组大鼠的创面组织TGF-β1和VEGF蛋白表达增加,NF-κB p65蛋白表达降低(P<0.05)。结论:猪脱细胞真皮基质可抑制大鼠瘘管创口组织炎症反应,促进血管新生及伤口愈合,且其促进胶原纤维生成的作用较瘘管栓更加明显,其机制可能与调控TGF-β1/NF-κB p65信号通路有关。

Objective:To observe the effect of acellular dermal matrix on the repair of fistula model in rats and on TGF-β1/NF-κB p65 signaling pathway,and to explore its possible repairing mechanism.Methods:After adaptive feeding for one week,40 healthy SD rats were randomly divided into four groups with 10 rats in each:sham operation group,model group,porcine acellular dermal matrix(pADM)group(dermal group),and positive control group(fistula plug group).After treatment for consecutive 2 weeks,the serum levels of IL-1β,IL-6,VEGF and prostaglandin E2(PGE2)were measured by ELISA.The wound tissues were collected to observe the pathological changes.Western blot was used to detect the expression levels of TGF-β1,NF-κB p65 and VEGF protein.Results:Compared with the model group,the dermal group and fistula plug group increased significantly(P<0.05)in the body weight,and the wound pus decreased significantly.The levels of serum IL-1β,IL-6 and PGE2 in the dermal group were lower and the serum level of VEGF was higher(P<0.05).Meanwhile,HE staining showed that inflammatory infiltration in the dermal group reduced obviously.Moreover,the collagen fibers increased significantly and became denser.In both dermal group and fistula plug group,the expression levels of TGF-β1 and VEGF proteins increased while the expression of NF-κB p65 protein decreased in wound tissues(P<0.05).Conclusion:Porcine acellular dermal matrix could inhibit the inflammatory reaction in fistula wound tissue in rats,promoting angiogenesis and wound healing.Its promoting effect on collagen generation is much more obvious in the dermal group than in the anal fistula plug group.The underlying mechanism might be related to the regulation of the TGF-β1/NF-κB p65 signaling pathway.