Mechanism Of Zihuang Shengji Ointment Promoting The Repair Of Chronic Infection Wounds By P38/MK2 Signaling Pathway

Objective:To investigate the repair effect of Zihuang Shengji Ointment on chronic infection wounds in rats,and to study the healing mechanism of Zihuang Shengji Ointment by the p38/MK2 signal pathway,so as to provide experimental basis for the clinical application of Zihuang Shengji Ointment.Methods:1.Consulting Fu Xiaobing.s and Zheng Xueping's building method,the model of rat chronic infection wound was established,which was randomly divided into normal group,model group,rb-bFGF group and Zihuang Shengji Ointment group.After the medication for 3d,7d and 14d observation time node,the wound healing rate,and histological observation of each rat were observed by HE staining,and the number of fiber cells and newborn blood capillary number were compared.2.The expression of TNF-a,IL-6,VEGF and Ang-1 proteins in the wound granulation tissues of rats were examined in the 3,7 and 14 days after the treatment detected by ELISA method,and the effects of Zihuang Shengji Ointment on the inflammatory response and angiogenesis of chronic infected wounds in rats were observed.3.The changes of MMP-1,MMP-2,MMP-9 and TIMP-1 in the wound granulation tissues of rats were examined in the 3,7 and 14 days after the treatment detected by IHC and Western Blot,and the intervention effects of purpura Zihuang Shengji Ointment on the expression of MMPs and TIMP-1 in the wounds of chronic infection were observed.4.The protein expressions of p38,p-p38,downstream MK2 and p-MK2,as well as the gene expressions of p38mRNA and MK2mRNA in the granulation of wound tissue of rats were examined in 3,7 and 14 days after treatment detected by Western Blot and RT-PCR,and the regulatory effect of Zihuang Shengji Ointment on the p38/MK2 signal pathway of chronic infection wound were observed.Results:1.Zihuang Shengji Ointment could improve the wound healing rate of chronic infection in rats,promoted the proliferation of wound fibroblasts and new capillaries,and shorten the wound healing time.2.Zihuang Shengji Ointment could down-regulate the expression of TNF-? and IL-6 in chronic infected wounds of rats and inhibit the inflammatory reaction of wounds.The expression of TNF-? and IL-6 in Zihuang Shengji Ointment group on the 3rd day after treatment were significantly lower than those in model group(P<0.01).The expression of TNF-? and IL-6 inZihuang Shengji Ointment group on the 7th and 14th day after treatment were different from those in model group and rb-bFGF group,and the differences were statistically significant(P<0.01).Zihuang Shengji Ointment could up-regulate the expression of VEGF and Ang-1 in chronic infected wounds of rats and promote the angiogenesis of wounds.The expression of VEGF and Ang-1 in Zihuang Shengji Ointment group were significantly higher than those in model group on the 3rd day after administration(P<0.01).The expression of VEGF and Ang-1 in Zihuang Shengji ointment group on the 7th and 14th day after administration were higher than those in model group and rb-bFGF group,and the differences were statistically significant(P<0.01).3.Zihuang Shengji Ointment could down-regulate the expression of MMP-1,MMP-2 and MMP-9,up-regulate the expression of TIMP-1,reduce the ratio of MMPs/TIMP-1,alleviate inflammation and accelerate wound epithelialization.The expression of MMP-1,MMP-2 and MMP-9 in Zihuang Shengji Ointment group were significantly lower than those in model group on the 3rd day after treatment(P<0.01).The expressions of MMP-1,MMP-2 and MMP-9 in Zihuang Shengji Ointment group on the 7th and 14th day after treatment were lower than those in model group(P<0.01),and the differences were statistically significant(P<0.01).The expression of TIMP-1 in Zihuang Shengji Ointment group were higher than that in model group on the 3rd day after treatment(P<0.01).The expression of TIMP-1 in Zihuang Shengji ointment group on the 7th and 14th day after treatment were significantly different from that in model group and rb-bFGF group,and the differences were statistically significant(P<0.01).4.Zihuang Shengji Ointment could inhibit the overexpression of p-p38,p-MK2 protein and p38mRNA,MK2mRNA gene,and improve the overexpression of p3 8/MK2 signal pathway.The expression of p-p38,p-MK2 protein and p38mRNA,MK2rnRNA gene in Zihuang Shengji Ointment group were significantly lower than those in model group on the 3rd day after administration(P<0.01).The expression of p-p38,p-MK2 protein and p38mRNA,MK2mRNA gene in Zihuang Shengji Ointment group on the 7th and 14th day after administration were lower than those in model group and rb-bFGF group,and the differences were statistically significant(P<0.01,P<0.05).Conclusion:Zihuang Shengji Ointment could inhibit the phosphorylation of p38 and downstream MK2,down-regulate the over-expression of p38 and downstream MK2 genes,improve the over-expression of p38/MK2 signaling pathway,regulate the expression of TNF-?,IL-6,MMPs and TIMP-1,and promote wound tissue repair.