紫朱软膏抑制PAD4介导的NETs形成促进糖尿病小鼠创面愈合的机制研究OA

Objective:To investigate the mechanism by which Zizhu Ointment(ZZO)promotes wound healing in diabetic mice by inhibiting neutrophil extracellular traps(NETs)mediated by peptidyl arginine deiminase 4(PAD4).Methods:A diabetic wound mouse model was used,and 48 male C57/BL6J mice were randomly divided into 4 groups:the normal wound group,the diabetic wound group,the ZZO treatment group,and the Beifuji treatment group,with 12 mice in each group.Wound healing was recorded by photography at different time points(0,3,7,and 14 days),and the differences in wound healing rates among the groups were analyzed.H&E staining was used to analyze the aggregation of inflammatory cells in the wound.Immunofluorescence was used to analyze the distribution of NETs in the wound.Western blot was used to detect the Cit-H3/H3 ratio in the wound.Im-munohistochemistry was used to detect the protein expression of PAD4,NOD-like receptor family pyrin domain containing 3(NL-RP3),and Interleukin-1β(IL-1β).RT-qPCR was used to detect the mRNA expression levels of PAD4,NLRP3,and IL-1β.Re-sults:The wound healing rate of the diabetic wound group was significantly lower than that of the normal wound group(P<0.01).The wound healing rate of the ZZO treatment group was significantly higher than that of the diabetic wound group(P<0.01),and the healing effect was better than that of the Beifuji treatment group on day 7(P<0.05).ZZO could inhibit the aggrega-tion of inflammatory cells(P<0.01)and the formation of NETs(P<0.01)in the wounds of diabetic mice,and reduce the protein expression(P<0.01)and mRNA levels(P<0.01)of PAD4,NLRP3,and IL-1β.Conclusion:ZZO may promote wound heal-ing in diabetic mice by inhibiting PAD4-mediated NETs formation and the release of inflammatory factors NLRP3 and IL-1β.