The Role And Mechanism Of Periplaneta Americana Adenosine Extract In Promoting Wound Healing

Objective:Rat full-thickness skin lesion model and skin repair-related cell line were used to evaluate the promoting effect of Periplaneta Americana adenosine extract(PAAE)on wound healing.Methods:Thirty SPF Sprague-Dawley(SD)rats were selected to establish a full-thickness skin lesion model,which was respectively pretreated with PAAE(12.5mg/ml),normal saline,and recombinant epidermal growth factor(10000IU/ml).At the beginning of the operation,the wound changes were recorded every other day.Take the 4th,6th,8th,10th,12th day of the wound and the surrounding 5mm skin was fixed and sliced.Using Hematoxylin-eosin and Mason trichrome staining,observing the thickness of fresh granulation tissue,collagen deposition and the degree of epithelial remodeling.Using histochemical staining,detection of TGF-β1 specific antigen expression in neonatal skin macrophages at different time points.Observing neovascularization by using immunofluorescence labeling of CD31 molecules.The effects of different concentrations of PAAE on the viability of human umbilical vein endothelial cells(HUVEC)and mouse monocyte macrophages(RAW264.7)were measured by MTT assay with tetramethylazozolium salt microenzyme reaction colorimetric assay(MTT).Western blot analysis was used to detect the changes of Smad2/3 protein expression in mouse monocyte-macrophage(RAW264.7)after different concentrations of PAAE.Scratch method was used to detect the migration of human skin fibroblasts(HSF)and human immortalized epidermal cells(HaCaT)after different concentrations of PAAE.Results:Compared with the saline group,the treatment of PAAE can accelerate the process of epithelial reconstruction and increase the formation of new blood vessels,and not increase the deposition of collagen fibers.MTT results showed that PAAE increased the activity of RAW264.7 and HUVEC cells in a concentration-dependent manner.Western Blotting suggested that treatment with PAAE could promote Smad2/3 phosphorylation.The scratch test demonstrated that the migration activity of HaCaT and HSF cells could be increased after treatment with PAAE.Conclusions:PAAE can accelerate the rate of epithelial remodeling after skin lesions and accelerate the repair of skin lesions.It was initially confirmed that PAAE is involved in the process of promoting skin lesion repair and the"macrophage-TGF-β1-Smad" is a key clue to wound repair.