Dnase1干预NETs对富血小板血浆加速糖尿病大鼠创面愈合相关机制的初步研究

Preliminary study on the related mechanism of Dnase1 interference of NETs on platelet-rich plasma accelerated wound healing in diabetic rats

目的探讨脱氧核糖核酸酶1(deoxyribonuclease 1,Dnase1)注射糖尿病大鼠创面降解中性粒细胞胞外诱捕网(neutrophil extracellular traps,NETs)后,再联合注射富血小板血浆(platelet-rich plasma,PRP)对创面愈合的影响.方法自2022年5-7月,华中科技大学协和深圳医院动物实验室联合拓普生物实验中心在糖尿病大鼠上建立4个溃疡创面模型,随机分为4组,6只/组.A组(Dnase+PRP组):创面边缘先后皮下点状注射Dnase1和PRP;B组(PRP组):创面边缘皮下注射PRP.C组(Dnase组):创面边缘皮下注射Dnase1;D组(对照组):创面边缘皮下注射生理盐水.分别于第5天、第14天各取12只大鼠,观察大鼠创面愈合程度、肉芽组织生长情况,切取创面组织,HE染色免疫组化法观察组织生长情况;检测和比较组织生长因子、炎性因子及NETs相关指标.结果治疗第5天后,各组创面愈合率无明显变化;治疗第14天后,A组创面愈合率最高,A组、B组创面血管内皮生长因子、转化生长因子-β1浓度高于C组、D组(P<0.05).A组、C组创面组织中髓过氧化物酶、弹性蛋白酶生成较B组、D组少(P<0.05).结论Dnase1和PRP联合注射,能够降低糖尿病大鼠创面NETs的水平,促进细胞增殖、血管新生,从而促进创面愈合.

Objective To investigate the impact of co-injection of deoxyribonuclease 1(Dnase1)and platelet-rich plasma(PRP)on wound healing in diabetic rats after degradation of neutrophil extracellular traps(NETs)by Dnase1 injection.Methods Four ulcer wound models were established in diabetic rats from May 2022 to July 2022,Huazhong University of Science and Technology Union Shen-zhen Hospital Animal Laboratory in collaboration with Topu Biological Experimental Center and randomly divided into 4 groups:Group A(Dnase+PRP):Dnase1 and PRP were sequentially injected subcutaneously at the edge of the wound.Group B(PRP):PRP was injected subcutaneously at the edge of the wound.Group C(Dnase):Dnase1 was injected subcutaneously at the edge of the wound.Group D(Con-trol):Normal saline was injected subcutaneously at the edge of the wound.Twelve rats from each group were selected on the 5th and 14th days to observe wound healing and granulation tissue growth.Wound tissues were collected,and histological examination using HE staining and immunohistochemistry was conducted to observe tissue growth.Tissue growth factors,inflammatory factors,and NETs-related indica-tors were detected and compared.Results After 5 days of treatment,there was no significant change in wound healing rate in each group.However,after 14 days of treatment,Group A showed the highest wound healing rate.The concentrations of vascular endothelial growth factor(VEGF)and transforming growth factor-β1(TGF-β1)in wound tissues of Group A and B were higher than those in the other two groups(P<0.05).The production of myeloperoxidase(MPO)and neutrophil elastase(NE)in wound tissues of Group A and C was lower than that in the other two groups(P<0.05).Conclusion Co-injection of Dnase1 and PRP can reduce the level of NETs in diabetic rat wounds,promote cell proliferation,angiogenesis,and thereby facilitate wound healing.