The Effect Of Inhibitor On The Skin Scar

Objective:To investigate the effects of arteannuin,artesunate,hirudin and margarita liquid on the proliferation of fibroblasts from human hypertrophic scar and the hypertrophic scars by cell invitro and animal experiments.It offered the theoretical and experimental evidences to further develop the effective external creams of inhibiting scars.Methods:1,The skin scars fibroblasts were cultured in vitro.This experiment were divided into serval groups:arteannuin group,artesunate group, hirudin group,margarita liquid group,ethanol group,sodium bicarbonate group and control group.MTT assay was used to examine the proliferation of the fibroblasts,then the inhibition rate was calculated.Apoptosis and its index(AI) in each group were determined after TUNEL staining and flow cytometry.2,Full-thickness wounds down to cartilage were created in the New Zealand white rabbits,for 12 scars each rabbit.Twenty-eight days after the initial wounding the hypertrophic scars on ears were begun with arteannuin,artesunate,hirudin and margarita liquid cream three times per day for 28 days. The untreated scars served as control scars and were left exposed to air.After 56 days of operation,the rabbits were killed,and the scars were harvested.The wound was fixed in 4%neutral-buffered formaldehyde,dehydrated,embedded in paraffin,cut in 4-μm sections,and stained with HE-stain and VG-stain.Then detected the hypertrophy index(HI)of a scar,the numerical density of fibroblasts on area(NA)and area density of collagen fibers on area(AA).Results:1,MTT assay rusult showed that margarita liquid could inhibit proliferation of fibroblasts derived from human scar in dose-dependent and duration dependent manner,the IC₅₀values of margarita liquid for fibroblasts was 15mg/L.Hirudin could inhibit proliferation of fibroblasts derived from human scar in dose-dependent and duration dependent manner,the IC₅₀values ofhirudin for fibroblasts was 0.9U/ml.Arteannuin could inhibit proliferation of fibroblasts derived from human scar in 0.206g/L and 0.103g/L.Artesunate could inhibit proliferation of fibroblasts derived from human scar in dose-dependent and duration dependent manner,the IC₅₀values of artesunate for fibroblasts was 263 mg/L.TUNEL staining rusult showed the inhibition rate in the experimental group with hirudin and margarita liquid were significantly higher than those in control groups(P＜0.01).And the inhibition rate in hirudin group was also obviously higher than margarita liquid group(P＜0.01).The apoptosis rate and the necrosis rate of the arteannuin group(0.206g/L)were higher than other two control groups(P＜0.01)。And the apoptosis rate and the necrosis rate in the group with artesunate(240mg/L and 120mg/L)were significantly higher than those in control groups(P＜0.01).2,Compared with control groups the arteannuin,artesunate and hirudin groups' scars apeared to be softer and flatter,the fibroblasts growed downward, the collagen fibers apeared to be regulated and sparse.There were obviously Statistical difference between control and experimental groups in the hypertrophy index(HI)of a scar,the numerical density of fibroblasts on area (NA)and area density of collagen fibers on area(AA).Conclusion:Arteannuin,artesunate,hirudin and margarita liquid could inhibit the proliferation of fibroblasts in skin scars.Artemisinin,artesunate and hirudin cream had practical potentialities in treatment and prevention of the animal studies skin scar hyperplasia.We will have a further research to investigate the optimization of combinating prescriptions in medicine of inhibiting human scars and observe the pharmacodynamic action of these medicines.