SangBai Hair Growth Formulas Influence In The Hair Regeneration Process Of The VEGF And Other Cytokines

Objective:Choose an old Chinese medicine prescription named SangBai Germinal Party as the studying object, the concept of different extracts SangBai Germinal Party on hair regeneration process of vascular endothelial growth factor (vascular endothelial growth factor, VEGF), interleukin-1β (interleukin-1 beta, IL-1β) and tumor necrosis factor a (tumor necrosis factor-alpha, TNF-α), and to explore its mechanisms of hair growth, provide a theoretical basis and experimental evidence for the clinical application of prescriptions.Methods:Complete scalp hair follicle transplantation occipital collected clinical surgery for isolation and culture of dermal papilla cells, dermal papilla cells take four to six generations orifice plate culture, divided into four groups:SangBai germinal water extraction liquid intervention group, SangBai germinal alcohol extract intervention group, positive control drug use 2% minoxidil, blank control group without drugs to stimulate; With tetramethyl azo thiazole blue (determined by MTT colorimetric determination of in vitro cultured human dermal papilla cells proliferation activity; Enzyme-linked immunosorbent assay (ELISA) to detect cells that IL-1 beta supernatant fluid, VEGF and the content of TNF alpha three kinds of cytokines; Reverse transcription polymerase chain reaction (RT-PCR) semi-quantitative detection of IL-1 beta cells, VEGF and TNF alpha mRNA expression; PI staining detected the dermal papilla cell cycle distribution flow; Using streaming Annexin/PI staining detection level changes between cell apoptosis; Using immunofluorescence flow cytometry to detect human dermal papilla cells IL-1 beta receptors, TNF alpha receptors and VEGF receptor expression levels.Results:Establish a perfect system of cultivating dermal papilla cells in vitro by determined by MTT test results from SangBai germinal side water extract(SBGW) stimulates the optimal concentration for cell proliferation is 0.7g/l and alcohol extract(SBGA) stimulates the optimal concentration for cell proliferation is 1g/1. ELISA results show that, compared with the blank control group, SBGW group and SBGA group can promote human dermal papilla cells secrete VEGF (P< 0.05, P< 0.01); Compared with positive control group, SBGW stimulates cells secrete VEGF and comparable (P> 0.05), SBGA group can significantly promote the human dermal papilla cells secrete VEGF (P< 0.05).Compared with the blank control group, SBGW can inhibit cell secretion of IL-1 beta (P< 0.05), SBGA and positive control group no significant effect of inhibiting the secretion of IL-1 beta (P> 0.05);Group compared with positive control group, SBGW could inhibit the secretion of IL-1 beta (P< 0.05), SBGA inhibition effect is not obvious (P> 0.05). Compared with the blank control group, both of SBGW and SBGA can promote the secretion and expression of TNF-alpha (P< 0.01, P< 0.05); Compared with positive control group, SBGW and SBGA has no statistical difference (P> 0.05). Rt-pcr results indicate:compared with the blank control group, SBGWcould promote the expression quantity of VEGFmRNA than the SBGA significantly (P< 0.01). Compared with positive control group, SBGA increase VEGFmRNA expression ability significantly (P< 0.01). Compared with the blank control group, both of SBGW and SBGA can inhibit IL-1 beta mRNA expression (P< 0.01); Compared with the positive control group, both the SBGW and SBGA inhibition of IL-1 beta mRNA expression was not significant (P> 0.05). Compared with the blank control group, the effect of SBGW to control theTNF-alpha mRNA expression was not significant (P> 0.05), SBGA can promote the expression of TNF-alpha mRNA (P< 0.05); Compared with positive control group, the difference of SBGW and SBGA was not significant (P> 0.05). Cell cycle and apoptosis detection results showed SangBai germinal extract can significantly reduce the apoptosis, help the dormant cells back into the growth period, the difference was statistically significant. Flow cell surface receptor experiment hints:compared with the blank control group, SBGW can promote the cell surface expression of VEGFR level (P< 0.05), SBGA and positive control group the levels of cell surface expression of VEGFR no difference (P> 0.05); Compared with positive control group, SBGW can obviously promote the cell surface VEGFR expression (P< 0.05), SBGA had no difference (P> 0.05);Compared with the blank group, SBGW and SBGA and positive control group both can inhibit human dermal papilla cells on the surface of the expression of IL-1 beta R (P< 0.05); Compared with positive control group, SBGW of human dermal papilla cells IL-1 beta receptors on the surface of inhibition, no statistical difference (P> 0.05); Compared with the blank control group, SBGA can promote human dermal papilla cells on the surface of the expression of TNF-alpha R (P< 0.05), SBGW and positive control group to the cell surface influence of TNF-alpha R ability is not obvious (P > 0.05); Compared with positive control group, SBGA significantly promote the expression of TNF-alpha R (P< 0.05), the compared with SBGW and positive control group has no statistical difference (P> 0.05).Conclusion:SangBai Germinal Party may be through the promotion of human dermal papilla cells synthesize and secrete VEGF, promote hair follicles to the synthesis of genetic material and proteins and inhibition of dermal papilla cells into early apoptosis to stimulate human dermal papilla cell proliferation, promote hair Freshmen role; by inhibiting the synthesis and secretion of inflammatory cytokines IL-1β to inhibit local inflammatory response, promoting the repair of damaged hair follicles effect; by promoting the synthesis and secretion of TNF-a to protect the role of the hair follicles.