Abstract:

Objective To investigate the effect of Chinese Herbal Anti-Alopecia and Hair Growth Solution （CHAaHGS） on the hair growth through in vitro experiments on the human dermal papilla cells （HDPCs）， in vivo experiments in the C57BL/6 mice， and human efficacy tests， and to clarify its potential mechanism. Methods The HDPCs were divided into control group， CHAaHGS group， and minoxidil group. MTT method was used to detect the proliferation activities of HDPCs in various groups； enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of vascular endothelial growth factor （VEGF）， hepatocyte growth factor （HGF）， insulin-like growth factor-1 （IGF-1）， and transforming growth factor β1 （TGF-β1） in the supernatant of HDPCs in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of VEGF， HGF， IGF-1， TGF-β1， and alkaline phosphatase （ALP） mRNA in the HDPCs in various groups； Western blotting method was used to detect the expression levels of β-catenin， dishevelled segment polarity protein 1 （DVL1）， glycogen synthase kinase 3β （GSK-3β）， phosphorylated GSK-3β （p-GSK-3β）， and wingless-type MMTV integration site family member 3a （Wnt3a） proteins in the HDPCs in various groups. A total of 18 mice were randomly divided into control group， CHAaHGS group， and minoxidil group， with 6 mice in each group. The mouse hair loss model was established using hair removal cream， and corresponding drug treatments were administered immediately after hair removal. The lengths and weights of newly grown hair on day 21 of the mice in various groups were detected； HE staining was used to observe the morphology of hair follicles in the dorsal depilated skin areas of the mice in various groups on day 7； ELISA method was used to detect the levels of VEGF， HGF， IGF-1， and TGF-β1 in the skin tissue of dorsal depilated areas of the mice in various groups. Sixty subjects were randomly divided into control group and CHAaHGS group， with 30 subjects in each group. The numbers of hair loss and hair densities of the subjects in various groups were detected at weeks 0， 4， 8， and 12. Results The MTT assay results showed that compared with control group， the proliferation activity of the cells in 50 mg·L^-1 CHAaHGS group was significantly increased （P<0.01）. The ELISA assay results showed that compared with control group， the levels of VEGF， HGF， and IGF-1 in the cell supernatant of HDPCs in CHAaHGS group were significantly increased （P<0.05 or P<0.01）， and the TGF-β1 level was significantly decreased （P<0.01）. The RT-qPCR results showed that compared with control group， the expression levels of VEGF， HGF， IGF-1， and ALP mRNA in the cells in CHAaHGS group were significantly increased （P<0.05 or P<0.01）， and the TGF-β1 mRNA expression level was significantly decreased （P<0.01）. The Western blotting results showed that compared with control group， the expression levels of β-catenin， DVL1， p-GSK-3β and Wnt3a proteins in the cells in CHAaHGS group were significantly increased （P<0.05 or P<0.01）， and the GSK-3β protein expression level was significantly decreased （P<0.05）. In animal experiments， on day 21， compared with control group， the length of newly grown hair of the mice in CHAaHGS group was significantly increased （P<0.05）， and the hair weight was significantly increased （P<0.01）. On day 7， the HE staining results showed that compared with control group， the hair follicle spacing of the mice in CHAaHGS group was significantly decreased （P<0.05）， and the number of hair follicles was significantly increased （P<0.01）； the ELISA assay results showed that compared with control group， the levels of VEGF， HGF， and IGF-1 in skin tissue of dorsal depilated area of the mice in CHAaHGS group were significantly increased （P<0.05 or P<0.01）， and the TGF-β1 level was significantly decreased （P<0.05）. In human efficacy test， compared with control group， the number of hair loss of the subjects in CHAaHGS group was significantly decreased at week 12 （P<0.01）， and the local hair density was increased （P<0.05）. Conclusion CHAaHGS promotes hair growth， and the mechanism may be related to its ability to increase the proliferation activity of HDPCs， induce the secretion of VEGF， HGF， and IGF-1， and activate the Wnt/β-catenin signaling pathway.

Key words: Chinese Herbal Anti-Alopecia and Hair Growth Solution, Human dermal papilla cell, Wnt/β-catenin signaling pathway, Hair growth, Minoxidil