Hair Growth Promoting Effect Of White Wax And Policosanol On The Mouse Model Of Testosterone Induced Hair Loss

Androgenetic alopecia(AGA)is more then seen in men compare to women.It is one of three common forms of hair loss.It can be begin at younger age with mental stress and dietary change,and it injure youger pepole self-respect.At present,finasteride and minoxidil are widely used to treat AGA.However,the two drugs have some adverse effects.Recently,natural products,which are safe and promote hair growth,have become widespread in the hair care industry.White wax(WW)is a secondary metabolite of the male larva of the white wax scale Ericerus pela Chavannes(Hemiptera: Coccoidea).In China,WW is used to treat hair loss.WW has been traditionally used to treat hair loss in China.However there has been no reporter WW responsible for hair growth-promoting effect.Policosanol of white wax(WWP)is a mixture of saturate monohydric alcohols,which can be prepared from WW with LiAIH4 and saponification.there was no report that hair-promoting effect of WWP.It have been reported that WWP possess extensive pharmacological activites.But there has been no reporter WWP responsible for hair growth-promoting effect.The present study was carried out to evaluate the effects of WW and WWP on testosterone-induced hair loss and the action mechanism in vivo and cell proliferation.In vivo,we employed visual inspection,morphology of hair follicle,level of hormone and growth factor to explore the change of follicle morphogenesis.In cell test,we employed human follicle dermal papilla cells(HFDPCs)as research object to evaluate cell proliferation in cultured HFDPCs and employed western blot to elvaluate protein expression in each group.The main results showed that:(1)The effect of WW and WWP on appearance of mice and morphology of hair follicleDarker hair and diffuse hair loss were visible in testosterone propionate only and testosterone propionate plus vehicle-treated groups.Follicles were shorten and miniaturization.However,in WW-treated groups and WWP-treated groups,Smooth and lustrous hair was saw.WW and WWP showed an outstanding hair growth-promoting activity as reflected by the follicular length,follicular density,A/T ratio,and hair bulb diameter.The optimal treatment effect was observed at 10% WW and 1% PPW,which were better than 2% finasteride treatment.(2)The adjustment of WW and WWP on hormoneThere were no effect of WW and WWP on estradiol level during test.But the level of testoserone and the ratio of testoserone and estradiol were change with the time.After 28 d,the levels of testosterone in mice treated with WW and WWP were decreased compare to model group,and the hormone levels was adjusted back to normal.But after 60 d,the ratio of testoserone and estradio was no difference between treatment group and CK groups,and the levels of testosterone in mice treated with 2%WWP was significantly lower than model group.Inhibitor assay of 5?-reductase showed that WW and WWP inhibited significantly the conversion of testosterone to dihydrotesterone,and the IC50 values of WW and WWP were higher than that of finasteride.(3)The effect of WW and WWP on growth factorsThe expression of VEGF and EGF,and the activity of ALP in mice treated with WW and WWP were significantly higher compare to model group.The level of VEGF and EGF was not relation to amouts.There were no differect between WW group or WWP group and 5% minoxidil for VEGF,in which there was no difference between WW group and WWP groups.Although there were no difference between in three concentration WW and WWP.the minimum level of EGF was 10% WW in WW group and the maximum level of EGF was 1% WWP in WWP group,and the expression of EGF in mice treated with 1%WWP was significantly higher than 10%WW group and 5% minoxidil,and there were no difference between 10%WW and 5% minoxidil.The activity of ALP was change with time.In which the tendency of ALP in 10%WW group and 1%WWP group were similar to 5% minoxidile.And the activity of ALP in 5%WW group and 20%WW group was difference between 28 d and 45 d,and then they decrease gradually.(4)The effect of WW and WWP on Human dermal papilla cellsTwain 80 have the properties to increase the solubility of WW and WWP.The optimistic solution conditions was : the ratio of twain80 and WW =50:1 or twain80 and WWP=5:1,pH 3~4,heating temperature is 91?,stirring time is 30 min.Human dermal papilla cells could grow well when the concentration of tween 80 was 1.56~6.25ug/mL.In the safe concentration range of tween80,white wax and policosanol from white wax had the cell proliferation effect for human dermal papilla cells and model cells.Werstern blot result showed that WW and WWP hold the capacity to inhibit the expression of TGF-??caspase3?caspase8 and caspase9.There were not differenct between white wax and policosanol for TGF-?.But the adjustment effect of caspase protein was obvious,in which white wax hold the higherproperty for caspase9,and policosanol hold the higher property for caspase3.In Conclusion,WW and WWP could act against testosterone-induced alopecia in mice,and they promoted hair growth.The action sites included hair bulb,5?-reductase and the complexes of dihydrotestosterone and the androgen receptor.The action mechanisim included prolong the anagenic phase of follicle and blocking the hormone to damage hair follicles.The property of prolong the anagenic phase of follicle for WW and WWP was related to promoting the expression of VEGF and EGF and DPCs proliferation.WW and WWP blocked the hormone to damage hair follicles by decreasing the level of testosterone and inhibiting 5?-reductase activity and inhibiting the complexes of dihydrotestosterone and the androgen receptor,and then inhibiting the caspase apoptosis signaling pathways,and in which white wax could inhibited the endogenous apoptosis signaling pathways,and policosanol could inhibited the exogenous signaling pathways.