The Study Of Actinidia Polygama Improving Erythema Of Sensitive Skin

Objective: Sensitive skin is a distinct skin type, attacking more and morepeople. It is disfined as the onset of pricking, buring, tingling, easilyassociated with erythema, papules, scaling, telangiectasia and orther objectsigns. Erythema is the most common symptom, seriously affect theappearance. The pathology showed the temporary telangiectasia showing upin papillary layer of dermis.In the skin, VEGF is mainly secrated by keratinocytes, including manyisoforms, can promoting vasodilation, leading to angiogenesis and increasingpermeability and other biological activities. Further studies have shown thaton the aspects of promoting vasodilation and increasing permeability,VEGF121is more significant than other isforms, maybe is the main factor thatcaused inflammatory erythema.Nitric oxide(NO) namely vascular permeability factor, exists widely inplant and animal. It as a signal conduction factor, exerting physiological andpathological functions, can promoting vasodilation, increasing vascularpermeability, is one of the most powerful vasodilation in the history. In thenormal skin, ketatinocytes, fibroblasts, endothelial cells also can produceNO.Endothelial NO is produced by oxidized L-arginine in nitric oxidesynthase. NO includs two subtypes,constitutive nitric oxide synthase (cNOS)and inducible nitric oxide synthase (iNOS). iNOS only substantially produce-ds in special cricumstance, eg: UV, TNF-α, IL-1β and other inflammatoryfactors, so it was emphasized pathological effect.Actinidia Polygama(AP) is a deciduous tuberous plant, its friuts, leaves,roots can be used as medicine. Studis show that AP has alkaloids, AP lactone,B-phenethyl alcohol and other ingredients, can treating many diseases. Korean researchers reported in2007that ALA in AP have anti-inflammtory effect,reducing vascular permeability, in perticular, can reduce swelling. Futherstudies show that ALA educing its anti-inflammtory effect through theinhibition of Nitric Oxide production and inducible nitric oxide synthase geneexpression via NF-κB and mitogen-activated protein kinase pathways.The study focuses on the typical manifestation-erythema of sensitive skin,mainly reasearching on VEGF closely related with vasodilatation. Our study isto explore whether AP can inhibit epidermal kerationcyte(EK) secretingVEGF; whether AP can antagnize VEGF121-induced vasodilatation and hyper-permeability through inhibiting NO production; whether AP can effect VEGF-121-induced the changes of intercelluar junction. The study can reveal themachanism of AP improving erythema from molecular biological level, so itcan provide a scientific basis for developing a new plant skin care product forsensitive skin.Method:1Extraction of effective composition from Actinidia Polygama2Research on AP effecting EK secreting VEGF using cells cultured in vitro.(1) Epdermal keratinocytes were cultured in vitro.(2) keratinocytes were stimulated by different concertration of the crudeextract of AP,24hours after, recover the medium and detect the VEGF inmedium with ELISA method. Choose the best concertration.(3) Analyze whether AP can inhibit kerationcytes secreting VEGF.3AP effect on NO production induced by VEGF121.(1) Human umbilical vein endothelial cells (HUVEC) were cultured invitro.(2) Use containing2%serum concertration as control.(3) HUVEC were stimulated by VEGF121, VEGF121+AP,2%serummediun.(4) Analyze whether AP can inhibit NO production using nitratereduction assay.4SPSS13.0software was used for statisitical analysis. All the data were represented as mean±standard deviation. Comparison of seveal means usingANOVA of randomized block design, different time points compared usingrepeated mesure analysis of variance, data of small sample size not meetingthe normality an homogeneity were analyzed using nonparametric test. Avalue of p<0.05was considered statistically significant.Results:1Effect of different concentrations of APW and APE on human epdermalkeratinocytes(HEK) secreting VEGF:(1) Detecting VEGF levels in medium stimulated by different concen-trations of APW and APE using ElISA method. The result showed that thereis a significant difference between the VEGF level sitimulated by differentcocentrations of APE and APW. Difference of the different concentrations of50μg/ml,100μg/ml,200μg/ml of APE respectively (0.353±0.069,0.183±0.015,0.183±0.015) comparing with the same dose of APW (0.353±0.069,0.183±0.015,0.183±0.015) has statistcally significant (p<0.05), no significantdifference when comparing control group (0.541±0.012)pg/ml (p<0.05), thatis VEGF level stimulated by different APE were significantly lower than APWand control group in a dose-dependent manner. There is no significantdifference between the different concentrations of APW, the matrix and thecontrol group(p>0.05)(2) Detecting cell viability stimulated by different concentrations ofdifferent extract using MTT assay: there is no difference between the differentextract methods and matrix on HEK, p>0.05, there is no difference betweenthe different concentrations of the extract, p>0.05, that is different concen-trations of AP extracts have no obviously cell toxicity on HEKs.2Effect of best concentration of APE on VEGF secreting by HEK in differenttime:Applying repeated measure, the VEGF level stimulated by the bestconcentration of APE has a significantly difference in different time(p<0.05).3The level of NO production secreting by HUVEC stimulated by VEGF121:Detecting NO production in medium using nitrate reduction assay: Thereis signifcant difference between the NO content secreting by HUVECs treated by the APE+of VEGF121group (3.263±0.367)μmol/ml and VEGF121group(5.017±0.660)μmol/ml(p<0.01), when APE+VEGF121group comparing tothe control group there has statistically difference(p<0.05), and no cytotoxicresponse(p>0.05), there is no signifcant difference when compared with1%matrix and the control group(p>0.05).Conclusion:AP ethanol(APE) extract has much stronger effluence on VEGF secretingby HEK than AP water extract in a dose-depended relationship. APE couldsignificantly inhibit VEGF121-induced human umbilical vein endothelial(HUVEC) secreting NO. Considering above all, we can conclude that APEcan improve the erythema of the sensitive skin by inhibition of the epidermalkeratinocytes secreting VEGF and partially antagonized VEGF121-inducedNO production excessively.