ABSTRACT
Objective To study the effects of Fructus ligustri lucidi and its monomers, tyrosol and oleanotic acid, on the migration of mouse melanoblast cell line (NCCmelb4M5). Methods Cultured NCCmelb4M5 cells were treated with Fructus ligustri lucidi (0.0625, 0.125, 0.25, 0.5, 2 mg/mL), tyrosol (0.02, 0.04, 0.08, 0.16, 0.8 mg/mL) and oleanolic acid (0.0625, 0.125, 0.25, 0.5, 2.5 mg/mL), respectively,for 48 hours followed by the detection of cell proliferation with MTT assay. The working concentration of the three drugs was determined according to the results of MTT assay. Scratch and transwell assays were performed to observe the effect of Fructus ligustri lucidi and its monomers at working concentration on the migration of NCCmelb4M5 cells. Results Based on the results of MTT assay, the working concentration of Fructus ligustri lucidi, tyrosol and oleanolic acid was determined at 0.125 mg/mL, 0.08 mg/mL and 0.0625 mg/mL respectively, and at these concentrations, these drugs exhibited a cytotoxity lower than that of absolute alcohol with no obvious stimulation of cell proliferation. Scratch and transwell assay revealed a promoting effect of both Fructus ligustri lucidi and tyrosol on melanoblast migration (P<0.05), while oleanolic acid had little effect on melanoblast migration. Conclusions The extract of Fructus ligustri lucidi has a significant stimulatory effect on the migration of mouse melanoblasts, and tyrosol may be an active component of Fructus ligustri lucidi associated with confirmative effect on migration of mouse melanoblasts.
ABSTRACT
Objective To investigate the level of NF-E2 related factor 2 (Nrf 2) in vitiligo lesions.Methods Tissue samples were obtained by press suction blisters at lesional and donor sites of 12 patients with vitiligo who were managed with epidermal transplantation. Four lesional samples from the patients were subjected to primary culture and the level of Nrf 2 was detected by AEC immunohistochemistry after 48hours of culture. Western blotting was utilized to further detect the level of cytoplasmic and nuclear Nrf 2 in tissue samples from the other 8 patients with vitiligo. Results Immunohistochemistry revealed that Nrf 2 was predominantly expressed in cytoplasm, rather than nuclei, of keratinocytes in vitiligo lesions compared with the normal skin of patients. The level of nuclear Nrf 2 was significantly lower in lesions than that in normal skin (0.10 ± 0.03 vs 0.26 ± 0.03, P ＜ 0.01) of the patients. In contrast, there was no significant dif- ference in the level of cytoplasmic Nrf2 between lesional and normal skin (0.61 ± 0.03 vs 0.60 ～ 0.02, P ＞0.05) of patients. Conclusion These results reveal an abnormality of nuclear translocation of Nrf 2 in vitili-go lesions.
ABSTRACT
Objective: To observe the therapeutic efficiency of adenovirus-mediated interleukin-12 gene(AdmIL-12) and CD40 ligand gene(AdmCD40L) intratumoral transfer in established murine melanoma in vivo. Methods: C57BL/6 mice were inoculated subcutaneously with B16 cells to establish the murine melanoma model. The tumor-bearing mice were injected intratumorally with murine IL-12 gene and CD40L gene recombinant adenovirus. Tumor growth and the survival of tumor-bearing mice were observed. The CTL activity was measured in vitro by lactate dehydrogenase(LDH) release assay. Results: Both AdmIL-12 and AdmCD40L can be efficiently expressed in vitro and in vivo. The treatment with AdmIL-12 could significantly inhibit the tumor growth and prolong the survival period of the tumor-beraing mice. Splenic CTL activity of the mice was also enhanced after IL-12 gene transfer. But the anti-tumor effects of AdmCD40L gene were not significant. In contrast, Co-delivery of IL-12 gene and CD40L gene lead to stronger antitumor effects than IL-12 gene alone. Conclusion: Adenovirus-mediated interleukin-12 gene and CD40 ligand gene transfer together intratumorally has significant therapeutic effects on mice melanoma in vivo.