Dracorhodin Perchlorate Regulate Collagen In Wound Healing

PURPOSE: Resina Draconis has the effect of promoting wound healing and hemostasis.Dracorhodin is a kind of flavonoid compound extracted from the Resina Draconis.It is unstable due to its existence in monomer form and is easil y reduced.Therefore,it is often present in the form of Dracorhodin Perchlorate.In this study,the Dracorhodin Perchlorate ointment was applied to the wound and its surrounding tissues to detect changes in collagen and its related factors,thereby studying the effect of Dracorhodin Perchlorate on collagen and regulation of proteins and their related factors in the wound healing process of SD rats.METHODS: Forty male SD rats were selected and weighed about 220 g.SD rats were randomly divided into model control group,and Dracorhodin Perchlorate was used to treat low,medium and high dose groups.There were 4 experimental groups,10 in each group.Rats were adapted for 7 days,and fasted for 12 h on the 7th day.The rats were anesthetized with 10% chloral hydrate at a dose of 0.35 m L/100 g.The back of the rat was then shaved and sterilized with 70% alcohol.Two full-thickness skin wounds were punched on the back of each rat using a sterile tissue punch with a diameter of 1 cm.After punching,the wound was disinfected with iodine.After the rats were observed to be anesthetized,they were routinely reared.The wounds of each group were treated with different concentrations of Dracorhodin Perchlorate ointment.The low-dose group ointment has a Dracorhodin Perchlorate content of 50 ?g/m L,and the middle-dose group ointment has a Dracorhodin Perchlorate content of 100 ?g/m L.In the high-dose group ointment,the Dracorhodin Perchlorate content was 200 ?g/m L,and the rats in the model control group were not treated,so that the wound naturally healed.The wound tissue of the rat skin was taken before modeling and on the 3rd,7th,10 th,and 14 th day after modeling.The expression and distribution of TGF-?1,MMP-2,MMP-9,TIMP-1 and TIMP-2 in the wound tissue of rats were detected by immunohistochemistry.The expressions of IL-1?,COX-2 and PDGF in the wound tissue of rats were detected by ELISA.Western blot was used to detect changes in the expression of TGF-?1,Smad2/3,p-Smad2/3,MMP-2,MMP-9,TIMP-1,TIMP-2,collagen ? and collagen ? in the healing skin tissue,RT-PCR detected changes in m RNA expression levels of MMP-2,MMP-9,TIMP-1,TIMP-2,collagen ?,and collagen ? in healing skin tissue.RESULTS: Compared with the model control group,the expression of collagen ? protein and m RNA in the healing tissue of rats was significantly increased from 0 to 14 d.The expression level reached the peak at 14 d;collagen ? at 0-7 d the expression of protein and m RNA increased,and the expression level of collagen ? decreased at 7 days.At 14 days,the expression level ofcollagen ? decreased to normal level.At 0-14 days,it could be the expression of TGF-?1 and Smad2/3 protein in the healing tissue of rats was significantly increased,and the phosphorylation level of Smad2/3 protein was significantly increased.At 0~14 d,DP ointment can significantly reduce the content of IL-1? and COX-2,while increasing the content of PDGF in rats wound healing skin tissue.The expression of MMP-9,TIMP-1 protein and m RNA were detected at 0~3 d.The amount was significantly increased,and the expression reached the maximum at the 3rd day,and the ratio of MMP-9/TIMP-1 was also the maximum.The expression of MMP-9,TIMP-1 and its m RNA at 3~14 d The ratio of MMP-9/TIMP-1 was also decreased,and the expression level returned to the normal level at 14 days,and the above effects were dose-dependent.At 3 days,the expression levels of MMP-2 and TIMP-2 protein and m RNA were not significantly different from those of the negative control group,but significantly increased compared wit h before the model establishment.At 3 to 14 days,MMP-2 and TIMP were observed.The expression of-2 protein and m RNA was not significantly different from that of the negative control group,and the ratio of MMP-2/TIMP-2 was not significantly changed.CONCLUSION: Dracorhodin Perchlorate can promote wound healing in rats.Wound healing can be promoted by increasing the amount of collagen expressed in the healing skin tissue.Dracorhodin Perchlorate promotes collagen synthesis by increasing the expression of TGF-?1,Smad2/3 protein and phosphorylation of Smad2/3 protein,and by reducing the expression of MMP-9 and by regulating the expression level of TIMP-1 inhibits the activity of MMP-9,thereby inhibiting the decomposition of collagen and achieving the purpose of promoting wound healing.