| Objective:By subcutaneous injection of neuropeptide substance P(SP), observe the SP influence on hair follicle cycle in C57 BL / 6 mice, To explore the role of SP in hair loss and possible mechanism.Methods:1. Using rosin and paraffin mixture to depilate the hair of C57 BL / 6 mice, establishment of animal model, in the back of the neck skin of mice by subcutaneous injection concentration of 10- 5 mol/L and 10-6 mol/L SP solution, respectively, after the injection, in the day of 1, 5, 9, 19 observation around the injection site mice skin color change;2. After depilation,Select the day of 9,19 point in time, measure the grey value of the picture around the injection site;3. In 9 and 19 days,take to the tissues of the mice skin at the injection site, HE staining to observe the changes of the hair follicle histological morphology, toluidine blue staining to observe the effects of SP of mast cell degranulation release, immune histochemical method detection CK19 and Nestin expression in tissue section.Results:1. The skin color changes of processing parts : by means of grey value determination, after injection of 9 days, control group mice at the injection site is black, 10-6SPgroup is dark grey, 10-5SP group as the light grey;19 days after injection, control group mice at the injection site for pink, 10-6SP group is light grey, 10-5SP group for dark gray;2.Compared with the normal group, nine days after depilation 10-6SPgroup, 10-5SP image grey value sp group were significantly higher, the difference was statistically significant(P < 0.05);19 days after hair removal, 10-5SP and 10-6SP group image of grey value significantly higher than the normal group, the difference was statistically significant(P < 0.05);3. HE staining crosscutting in each hair follicle number comparison: 9 days after hair removal, compared with normal group, 10-6 SP and 10-5 SP groups in the number of follicles significantly reduced, the difference was statistically significant(P < 0.05);19 days after hair removal,compared with normal group, 10-6 SP group and the number of follicles in 10-5SP group increased significantly, the difference was statistically significant(P < 0.05);4. Number of mast cells degranulation comparison: 9 and 19 days after hair removal, 10-5SP and 10-6SP groups of local visible mast cell degranulation phenomenon, and 10-5SP groups than 10-6SP group had a significantly increased the number of MCS degranulation, the difference was statistically significant(P < 0.05);The normal group of mast cells in crumb, almost no degranulation phenomenon. 5. Immunohistochemical expression: after hair depilation in 9 days, in the normal group, 10-6SP, 10-5SP groups can all find CK19 positive cells in hair follicle outer root sheath, sweat glands, and the base layer were, and there was no evident difference between groups of positive expression of CK19 and it no statistical significance(P > 0.05);Nestin positive cells appeared in the three groups of skin epidermis, hair follicles, sebaceous glands, and part of the sweat glands, the positive expression rate had no statistical significance(P > 0.05);19 days, CK19 were and the position of Nestin positive cells appeared and 9 days, the same group of CK19 were and Nestin positive expression rate had no statistical significance(P > 0.05).Conclusion:1. After subcutaneous injection of SP solution can affect the normal progress of the hair follicles cycle in mice, showing that it can slow the growth of hair follicles at anagen.2. SP can make the mast cell around the hair follicle produce degranulation phenomenon,and is associated with concentration of SP. 3. SP- MC pathway may be involved in the regulation of hair follicle cycle in mice.4. SP had no effect on the hair follicle stem cell markers of CK19 and Nestin. |
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