Mechanism and Safety Study of External Use of Kuiyangsan on Wound Healing for Skin Ulcer in Rats
Objective To study the mechanism and safety of external use of Kuiyangsan(KYS)on wound healing of skin ulcer in rats.Methods A total of 110 SPF Sprague-Dawley rats were divided into 2 groups by random number table.Twenty rats used for safety experiment were randomly divided into the complete skin group(n=10)and the damaged skin group(n=10),and the other 90 were divided into the control group(n=15),the model group(n=15),the rh-bFGF group(n=30),and the KYS group(n=30)by random number table.An skin ulcer wound was established on the back of rats'feet by a high-fat high-sugar diet combined with intraperitoneal injection of streptozotocin(STZ).Recombinant human-basic fibroblast growth factor(rh-bFGF,150 IU/cm2)was externally administered to rats in the rh-bFGF group.KYS(1 g/cm2)was externally applied to rats in the KYS group.Wound healing was observed to calculate the wound healing rate.Sections of the wound granulation tissue were collected for HE staining and histopathological observation before treatment,and at day 7 and 14 after treatment.The gene expressions of tumor growth factor β 1(TGF-β 1)and Smad3 in the wound granulation tissue were detected by PCR.The expression of cleaved caspase-3 was determined by immunohistochemistry.The TUNEL Apoptosis Detection Kit was used to detect the apoptosis of tissue cells.In the safety experiments 20 healthy male Sprague-Dawley rats were shaved on both sides of the back spine,and the damaged skin group was rubbed with fine sandpaper to cause local abrasion in the shaved area.The left and right sides of the shaved area were coated with KYS and Vaseline respectively.Skin irritation was observed,urine test and β2-MG level test were performed.Results Compared with the control group,the wound healing rate significantly decreased in the model group at day 7 and 14 after administration(P＜0.01).Inflammatory cell infiltration and few new capillaries and few collagen fibers were observed.Compared with the model group,the wound healing rate of the KYS group and the rh-bFGF group significantly increased(P＜0.01).There were more fibroblasts and less infiltration of inflammatory cells in the wound tissue,indicating good repair.mRNA expression levels of TGF-β 1 and Smad3 significantly increased(P＜0.01).The cleaved caspase-3 protein expression level significantly decreased(P＜0.01).The degree of apoptosis in the wound surface significantly decreased(P＜0.01).In the safety experiments there were no significant differences in the degree of skin irritation,24 h albumin(ALB)levels,or β 2-MG levels between the two groups(P＞0.05).Conclusion KYS accelerated neovasculogenesis and collagen fiber deposition,reduced apoptosis,and promoted wound healing in rats by TGF-β 1/Smad3 signaling pathway,and it had a remarkable safety profile.
external use of Kuiyangsanskin ulcertransforming growth factor-β 1/Smad3 signaling pathwaywound healingChinese herbal compound
国家科技期刊平台
NSTL
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