The Effects Of Different Concentrations Dihydrotestosterone On Human Dermal Papilla Cells,Wnt And Apoptosis Pathways

Objective: To clarify the correlation between DHT and the growth of human dermal papilla cells and investigate the mechanism by observing the effect of different concentration Dihydrotestosterone(DHT)on human dermal papilla cell,providing a scientific basis for the design of the new prevention and treatment of androgen alopecia..Methods: the control group and the dihydrotestosterone groups with concentrations of 0.1ng/ml,1ng/ml,10ng/ml,100ng/ml and 1000ng/ml were respectively co-cultured with human hair papilla cells.The hair papilla cells were identified by ?-SMA immunofluorescence technology.The proliferation and apoptosis rate of human hair papilla cells were determined by MTT assay and flow cytometry.RNA-seq was used to detect gene expression in the Wnt and apoptosis pathway.Comparative and correlation analysis was conducted by SPSS 23.0.Results:(1)immunofluorescence: distribution density of dermal pailla cells in control group and 0.1 ng/ml,100 ng/ml,1000 ng/ml DHT group is sparser than which in the 1 ng/ml and 10 ng/ml DHT group,and the cells in 10 ng/ml DHT group are most densest.(2)MTT assay: compared with the control group,0.1ng/ml DHT can not promote the proliferation of hair papilla cells(P?0.05),while 1ng/ml and 10ng/ml DHT promoted the proliferation of hair papilla cells(P?0.01,P?0.01).There is no inhibitory effect on proliferation of hairy papilla cells in the group with 100ng/ml DHT(P?0.05),while 1000ng/ml DHT inhibite the proliferation of hairy papilla cells(P?0.05).(3)flow cytometry: 0.1ng/ml,1ng/ml and 100ng/ml DHT do not promote the apoptosis activity of DPCs(P>0.05).1000ng/ml DHT can promote apoptosis of DPCs(P?0.01).(4)The effect of DHT on Wnt pathway was detected by RNA-seq: low concentration DHT(?10ng/ml)up-regulates the expression of genes Wnt10 b and LEF1,and down-regulates the expression of genes Axin2,GSK-3?and DKK-1.high concentration DHT(>100ng/ml)up-regulates the expression of genes Axin2,GSK-3?and DKK-1,and down-regulates the expression of genes Wnt10 b and LEF1.Effects of DHT on apoptosis pathway detected by RNA-seq assay: low concentration DHT(?10ng/ml)up-regulates Bcl-2 gene expression,down-regulates Bax and Fas gene expression.high concentration of DHT(> 100ng/ml)up-regulates Bax and Fas gene expression and down-regulates bcl-2 gene expression.Conclusion: 1.DHT promotes the proliferation of dermal papilla cells at a certain concentration(1ng/ml,10ng/ml),high concentration DHT(1000ng/ml)inhibites the proliferation of dermal papilla cells and promotes the apoptosis.2.Different concentrations of DHT can influence the signaling factors in the Wnt and apoptosis pathway,and affecting the proliferation and apoptosis of human dermal papilla cells.