Study Of The Role Of SCF/KIT Pathway In The Treatment Of Vitiligo With Monochromatic Excimer Light

Objective To study the role of stem cell factor(SCF)/KIT pathway in the pathogenesis of vitiligo. Methods The expression of SCF and KIT protein was detected by immunohistochemistry in 12 patients with trichromatic vitiligo. The enzyme-linked immunosorbentassay(ELISA) was used to detect the concentration of soluble SCF in the fluid of blister in 25 patients with vitiligo in stable stage. Results Compared with the non-lesional epidermis of trichromatic vitiligo, the expression of SCF protein was higher in the epidermis of milky white and white lesion, and that in the latter is more higher. In contrast with the expression of SCF, a lower protein expression of KIT was detected in the epidermis of milky white lesion, and in the white lesion non KIT protein was detected. The concentration of soluble SCF was higher in the lesional blister fluid of the patients with stable vitiligo than that in the non-lesional blister fluid, and the expression of soluble SCF was related to the recovery degrees after the whole epidermis transplantation. Conclusion The abnormal expression of SCF and KIT may be involved in the pathogenesis of vitiligo. And soluble SCF may be play a great role in the recovery of vitiligo lesions.中国分类号:R75PartⅡEffects of 308 nm monochromatic excimer light on cell characteristicsand SCF expression of human keratinocytesObjective To investigate the effect of 308 nm monochromatic excimer light(MEL) on cell characteristics and SCF expression of human keratinocytes. Methods We used MTT to determine the suitable dose of 308nm MEL on the keratinocytes, and flow cytometry was used to examine the effect of 308 nm MEL on the cell apoptosis and cell cycle. The mRNA and protein expression of SCF was detected by RT-PCR and Western blotting and ELISA, respectively. Results 308 nm MEL induced cell apoptosis at certain doses(p<0．05) , but significantly up-regulated SCF mRNA and protein expression, and protein expression increased with the increasing doses of 308 nm MEL. There was no differece in the mRNA expression in different doses groups though they were highei than that in the control. Conclusion 308 nm MEL induced apoptosis of human keratinocytes, but promoted the expression of SCF mRNA and protein, which might be the one of mechanisms of 308 nm MEL in the treatment of vitiligo.中图分类号: R75PartⅢEffects of 308 nm monochromatic excimer light on cell characteristics and KIT expression of human melanocytesObjective To evaluate the effect of 308 nm monochromatic excimer light(MEL) on cell apoptosis, cell cycle and KIT expression of human melanocytes. Methods MTT was used to detect the cell viability of melanocytes irradiated by 308nm MEL with low and high doses after 24h. Annexin-V with flow cytometry was used to examine the early apoptosis of melanocytes exposed to 308nm MEL with different doses after 24h, and PI with flow cytometry was used to detect the cell cycle of melanocytes irradiated by 308nm MEL with 80mJ/cm~2 and 400mJ/cm~2 after 24h and 48h. After exposed to 308nm MEL 24h, Real time RT-PCR and western-blot were used to detect the expression of mRNA and protein of c-kit in melanocytes, respectively. Results There was no difference of cell viability in melanocytes exposed to 308nm MEL with low doses between irradiaed cells and the control. But the cell viability decreased significantly after irradiated by 308nm MEL at dose of 600mJ/cm~2 (P＜0．05) . At the low doses group, 308nm MEL showed the tendency to inhibit the cell apoptosis, but there was no statistics significance compared to the control. The cell apoptosis increased obviously in melanocytes irradiated by 308nm MEL at the dose of 400mJ/cm~2 after 24h. Exposed to 308nm MEL at the dose of 80mJ/cm~2, the percentage of melanocytes in S phage was 29．06±0．75% and 12．28±0．82% at 24h and 48h, respectively, both were higher than that in the control(P<0．05) . However at the dose of 400 mJ/cm~2 , the percentage of melanocytes in S phage were both lower than that in the control(P<0．05) . The expression of mRNA and protein of c-kit were both upregulated at the low doses group, and to achieve the peak at the dose of 80mJ/cm~2．However, 308nm MEL at the dose of 400mJ/cm~2 inhibited significantly the expression of mRNA and protein of c-kit in 4 hours. Conclusion 308nm MEL promotes the proliferation and the expression of KIT protein of melanocytes at low doses ,which may be one of the mechanisms of 308nm MEL in the treatment of vitiligo. And at high doses, 308nm MEL inhibits the proliferation and the expression of KIT protein of melanocytes, which may be involved in the pathogenesis of vitiligo patients while exposed to strong sun light.中图分类号: R75PartⅣEffect of the co-culture of keratinocytes and melanocytes and soluble SCF on cell proliferation, melanogenesis and KIT expression of melanocytesObjective To investigate the effect of soluble SCF on cell proliferation, melanogenesis and KIT expression in melanocytes. Methods The soluble SCF was added into the melanocytes culture medium (50ng/ml) and the melanocyte and keratinocyte were co-cultured as the control. The c-kit mRNA and protein expression were detected by Real-time PCR and western-blot, respectively at 4h and 24h after the soluble SCF was added into the medium. The NaoH-assay and cell count were used to detected the melanogenesis and proliferation of melanocytes, respecrively. Results There was no difference of the c-kit mRNA expession in melanocytes between the co-culture of melanocytes and keratinocytes and the melanocytes added with soluble SCF at 4h and 24h (P>0．05) . But the expression of KIT protein decreased at 4h and incteased at 24h in the two groups compared to the control. And at 72h after the soluble SCF was added, the number of melanocytes was increased in the sSCF group compared to the control(P<0．05) , but the melanogenesis of melanocytes increased only in the co-culture of melanocytes and keranocytes(P<0．05) . Conclusion The KIT protein may be neutralized by the soluble SCF which stimulated the proliferation but the melanogenesis of melanocytes.中图分类号:R75...