Anti-fibrosis Of Capparis Spinosa Extracts In Animal Models Of Scleroderma

BackgroundScleroderma is a connective tissue disease characterized by fibrotic skin or internal organs, developing to atrophy finally. Scleroderma has two major clinical subtypes:limited and diffuse. Now,the incidence of scleroderma is in the third of connective tissue diseases which only after rheumatoid arthritis and lupus erythematosus in China. The exact etiology and pathogenesis of scleroderma remains unknown. In currently, there are no ideal methods and medicines to heal it,if the important organs involved,such as pulmonary, cardiac, renal, gastrointestinal, the prognosis is poor even lead to death. Limited SSc is characterized by fibrotic skin changes that are limited to the fingers, hands,face and other exposure portions which influence the beauty seriously. Therefore, the reasonable and effective treatment of scleroderma is very important.The fibroblast is an important cell to study in SSc because of the complex extracellular matrix changes seen in this disease. SSc fibroblasts produce more collagen type Ⅰ,Ⅲ, Ⅵ, Ⅶ than normal, especially collagen type Ⅰ. In the process, beta transforming growth factor (TGF-beta) is an important factor to stimulate fibroblast synthesis of type Ⅰ collagen.Attention has focused on transforming growth factor-b(TGF-b) and more recently,on connective tissue growth factor (CTGF). The latter induced by TGF-beta may be responsible for sustained stimulation of collagen synthesis. CTGF is a necessary factor to maintain fibrosis, dominating scleroderma fibrosis pathological process with TGF-b.Capparis spinosa L. also named caper, has the efficacy of dispelling wind and eliminating dampness,analgesia,activating blood ciuculation,detumescence and so on.As uighur medicine in Xinjiang province this plant has widely use. Previous chemical studies on Capparis spinosa L.have shown the presence of alkaloids,lipids,polyphenols,flavonoids,indole and aliphatic glucosinolates.These flavonoids display a remarkable role in various pharmacological activities including antifungal,anti-inflammatory,antiallergic,antihepatotoxic and anti-diabetic effects.The methanol extract of C.spinosa flower bud(LECS) showed a noteworth antioxidant/free radical scavenging effectiveness in diverse in vitro test. Topical used LECS could distinctly relieve the erythema induced by UVB in vivo test. The LECS could significantly inhibit inflammation media caused by IL-1beta,such as the release of nitric oxide,prostaglandin E2and reactive oxygen species(ROS), hence, we speculate C.spinosa posses a noteworthy anti-fibrotic, antioxidant effectiveness. We did some experiments in vitro study and found the ethanol extracts of C.spinosa fruit on the expression of type Ⅰ collagen in scleroderma patient,but for the normal person, fibroblast morphology had no obviously change after dosing.Apoptosis, dying cells had no distinctly increase. These prompted ethanol extract of C.spinosa had the effects of selective anti fibrosis of skin and was superior to Centella Triterpenes Cream. We had select the effective parts.On the basis of the previous studies, we designed this experiment by establishing the C3H mice model of scleroderma by bleomycin. Observed the thickness of derma,collagen fibrous tissue chemical index, hydroxyproline content in the skin of different concentration Capparis spinosa extracts ointments and Centella Triterpenes Cream. Confirming effective parts of C.spinosa in vivo.In previous studies,we had found the effective parts through extraction technology research, pharmacodynamic experiment,which made it easy to industrialized production,we also got the stable quality preparation through preparation technology, quality standard, stability study. The effective parts preparation of C.spinosa in the treatment in vitro, exploring new areas for the treatment of scleroderma and providing the basis for further development and application of C.spinosa. ObjectiveTo investigate the anti-fibrosis of Capparis spinosa extracts in animal models of scleroderma, filtrating the optimal concentration extract ointment.Materials and methodsFemale healthy, C3H mice (N=54) were equally and randomly divided into six goups, shaved off the hair of central back, area1.5cm*2cm.C3H Mouse models of scleroderma were established through local injection of bleomycin on the back once a day for3weeks.From the first day,treatment groups external application ointment.Three groups respectively to received topical treatment with1%,2.5%,5%ethanol extract of C.spinosa.One group was received topical treatment with Centella Triterpenes Cream,no treatment was given to the rest of the control group. After3-week treatment,54mice were sacrificed and target area of the skin was obtained from the back,and subjected to the measurement of dermal thickness by HE staining, collagen fibrous tissue chemical index by Masson three-colour staining and hydroxyproline content by Hydroxyproline detection kit.Statistical analysisAll data were managed with SPSS20.0for windows.,the datas were described with mean±SD. Student’s t test, One-Way ANOVA analysis were used. P<0.05was considered to be statistically significant in all cases.Results1. The thickness of derma The target area of the skin of bleomycin group was thicker significantly than normal control group(P<0.05),1%,2.5%, ethanol extract of C.spinosa groups could inhibit dermal thickness obviously induced by bleomycin, especially2.5%group,5%group, Centella Triterpenes Cream group was weaker than the former two(P<0.05).2. Collagen fibrous tissue chemical index Collagen fibers of the target area of the skin of bleomycin group increased, thicked distinctly, lined up tightly, collagen fibrous tissue chemical index was higher obviously than normal control group(P<0.05).1%,2.5%,groups could significantly inhibit the expression of collagen fibers in dermis, collagen fibrous tissue chemical index of these groups were below the bleomycin group (P<0.05), the expression of collagen fibers of Centella Triterpenes Cream group was weaker than C.spinosa groups.significantly.3. Hydroxyproline content The content of hydroxyproline of bleomycin group increased obviously than normal control group(P<0.05), C.spinosa groups were lower than bleomycin group. Centella Triterpenes Cream group was between bleomycin group and C.spinosa groups.ConclusionThe ethanol extracts of Capparis spinosa effectively inhibit skin sclerosis, the concentration of1%,2.5%,groups are superior to Centella Triterpenes Cream group, with no irritation.