| Dracorhodin perchlorate was a chemical synthesis analogue of the blood residue,which was the stable state of the active ingredient in blood exhaustion.Dragon'blood was widely used in clinical treatment of fracture,trauma,inflammation and other related diseases,but the effect of the active ingredient Dp on wound treatment was rare reported.In order to investigate the effect and mechanism of Dp on skin wound healing,60 Wister rats were used and the weight was about 250 g.A model of skin defect was prepared in 60 male rats,and the traumatic model rats were randomly divided into model group and drug group,each group had 30 rats.Model group Smear Vaseline,the drug group smear from Vaseline for the substrate prepared by the Dp ointment for drug treatment,at 3,7,10,14,21 d,6 rats were randomly taken to take photos of the wound and Image Pro Plus analysis software was used for wound healing rate statistics analysis;The rats were executed by the method of taking off vertebra,and the skin tissue around 2 cm was taken.The degree of inflammatory cell infiltration and the number of microvascular regeneration were detected by the staining and CD34 immunohistochemical staining in rats after Dp treatment.Masson tissue staining was used to detect collagen synthesis in wound.The expression levels of inflammatory factors such as interleukin-1?(IL-1?)and tumor necrosis factor(TNF-?)were detected by enzyme-linked immunosorbent assay(ELISA),and the transforming growth factor-?(TGF-?)and vascular endothelial growth factor(VEGF)were detected by Western blot assay.The expression levels of hepatocyte growth factor(HGF),fibroblast growth factor(FGF-7)and other genes were detected by fluorescence quantitative PCR assay.Results: The wound healing rate showed that DP could promote wound closure(p<0.05)more quickly at 7 d and 10 d.He staining showed that the inflammatory cells in the wound tissues of the drug group and the model group increased significantly at 3 d,but the number of inflammatory cells in the drug group was lower than the model group(p<0.05),and the skin healing of the drug group was more flat than the control group.Immunohistochemical staining of CD-34 showed that the number of microvessels in the DP group was significantly higher than the model group(p<0.05)at 3d and 7d.Masson staining showed that DP group could promote the deposition of wound collagen,the data showed that there was no significant difference in collagen content between the model group and the drug group at 3 d,and the collagen content density of model group and DP Group was 53% and 68%(p<0.05)respectively at 7 d,and 65% and 79% respectively in 14 d,the differences was significant(p<0.05).Detection of inflammatory factor expression in rat skin tissue by ELISA kits showed that the level of IL-1? secretion in DP group was significantly lower than that in the blank dose group(p<0.05)in the 3 d,and the TNF-? secretion level was significantly lower than that in the blank dose group(p<0.05)at 3 d and 7 d.The secretion level of MPO and PGE synthase was significantly higher than normal level in the 3 and 7 d,but there was no significant difference between the drug group and the control group.The level of protein expression showed no statistically significant difference in protein expression between the blank group and the drug group at 3d,but the expression of TGF-? protein in the drug group was significantly higher than that of the control group(p<0.05)at 7 d of wound healing,and the expression of VEGF protein was opposite to that of TGF-? on 3 d,The expression of VEGF protein in drug group was higher than control group,and the differences was statistical(p<0.05).The results of fluorescence quantitative PCR showed no significant difference in HGF gene expression between DP group and control group.The expression of EGF was significantly higher than that in the control Group(p<0.05)in 3 d after the trauma occurred in the body.The expression of FGF-7 increased with the healing time,and there was no significant difference between the drug group and the control group at 3 d,and the FGF-7 gene expression of DP group was significantly higher than the blank group(p<0.05).Conclusion: DP promotes skin wound healing.The main mechanism is to restrain the inflammatory reaction of the wound,weaken the inflammatory infiltration of the wound,inhibit the expression of the related inflammatory factors such as IL-1? and TNF-?.At the same time,DP can promote the formation of microvascular and the synthesis of collagen,and regulate the expression of related growth factors to promote the rate of wound healing. |
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