富血小板血浆促糖尿病创面愈合机制的初步研究

Mechanism of platelet-rich plasma in promotion of diabetic wound healing:a preliminary study

目的探讨富血小板血浆(PRP)促进糖尿病创面愈合与核苷酸结合寡聚化结构域样受体蛋白(NLRP3)炎性反应复合物/IL-1β信号通路的关系。方法共收集25份组织标本,5份取自5例糖尿病截肢患者截下肢体的远端坏死交界区(DMD组),5份取自前述5例糖尿病截肢患者截下肢体的近端创面(DMP组),5份取自5例非糖尿病截肢患者截下肢体的近端创面(NDM组),5份取自5例糖尿病足溃疡保肢治疗患者的溃疡创面使用PRP治疗前行清创时去除的组织(NPRP组),5份取自前述5例糖尿病足溃疡保肢治疗患者的溃疡创面使用PRP治疗后首次行清创时去除的组织(PRP组)。分别采用Western印迹法检测NIRP3蛋白表达,ELISA检测IL-1β蛋白表达,实时荧光定量PCR检测NLRP3基因和IL-1β基因表达。结果 DMD组和DMP组的IL广1β和NLRP3蛋白相对表达量和基因相对表达量均显著高于NDM组(P值均<0.05),DMP组的IL-1β和NIRP3蛋白相对表达量和基因表达量均显著高于DMD组(P值均<0.05)。NPRP组的IL-1β和NLRP3蛋白相对表达量和基因相对表达量均显著高于PRP组(P值均<0.05)。结论 NLRP3炎性反应复合物/IL-1β信号通路上调可阻碍糖尿病创面的愈合。PRP通过抑制NLRP3炎性反应复合物/IL-1β信号通路,加快糖尿病创面愈合进程,是其促进糖尿病创面愈合的机制之一。

Objective To explore the relationship between platelet-rich plasma （PRP） and nod-like receptor protein 3 （NLRP3） inflammasome/ interleukine-1β （IL-16） signaling pathway in the process of diabetic wound healing. Methods A total of 25 tissue samples were collected in this study, including 5 from distal necrosis border areas of amputated limbs of 5 patients with severe diabetic foot ulcer （DMD group）, 5 from the proximal wound of amputated limbs of these patients （DMP group）, 5 from proximal wound tissue of amputated limbs of 5 non-diabetic patients undergoing amputation treatment （NDM group）, 5 from wound tissue in the first debridement before use of PRP in 5 patients with mild diabetic foot who had limb-salvage indications （NPRP group）, and 5 from wound tissue in the next debridement after PRP therapy （PRP group）. The protein expressions of NLRP3 and IL-1β were detected through Western blotting and enzyme linked immunosorbent assay （ELISA）. The mRNA levels of NLRP3 and IL-1β were measured by real-time polymerase chain reaction （RT-PCR）. Results Both protein and mRNA expression of IL-1β and NLRP3 in DMD group and DMP group were significantly higher than those in NDM group （all P〈0.05）. Meanwhile, protein and mRNA expression of IL-1β and NLRP3 in DMP group were significantly higher than those in DMD group （all P〈0.05）. Both protein and mRNA expression of IL-1β and NLRP3 in NPRP group were significantly higher than those in PRP group （all P〈0. 05）. Conclusion The up-regulation of NLRP3 inflammasome/IL-1β signaling pathway may hinder diabetic wound healing. PRP can accelerate the process of diabetic wound healing by inhibiting NLRP3 inflammasome/IL-1β signaling pathway. （Shanghai Med J, 2017, 40： 169-172）