ABSTRACT
OBJECTIVE@#To evaluate the efficacy of rapmycin for treatment of experimental autoimmune encephalomyelitis (EAE) in mice and explore the underlying mechanism.@*METHODS@#An EAE model was established in C57BL/6 mice. After immunization, the mice were divided into model group and rapamycin groups treated daily with low-dose (0.3 mg/kg) or high-dose (1 mg/kg) rapamycin. The clinical scores of the mice were observed using Knoz score, the infiltration of IL-17 cells in the central nervous system (CNS) was determined using immunohistochemistry; the differentiation of peripheral Treg cells was analyzed using flow cytometry, and the changes in the levels of cytokines were detected with ELISA; the changes in the expressions of p-Smad2 and p- smad3 were investigated using Western blotting.@*RESULTS@#High-dose rapamycin significantly improved the neurological deficits scores of EAE mice. In high-dose rapamycin group, the scores in the onset stage, peak stage and remission stage were 0.14±0.38, 0.43±1.13 and 0.14±0.37, respectively, as compared with 1.14±0.69, 2.14±1.06 and 2.2±0.75 in the model group. The infiltration of inflammatory IL-17 cells was significantly lower in high-dose rapamycin group than in the model group (43±1.83 153.5±7.02). High-dose rapamycin obviously inhibited the production of IL-12, IFN-γ, IL-17 and IL-23 and induced the anti-inflammatory cytokines IL-10 and TGF-β. The percentage of Treg in CD4+ T cells was significantly higher in high- dose rapamycin group than in the model group (10.17 ± 0.68 3.52 ± 0.32). In the experiment, combined treatments of the lymphocytes isolated from the mice with rapamycin and TGF-β induced a significant increase in the number of Treg cells (13.66±1.89) compared with the treatment with rapamycin (6.23±0.80) or TGF-β (4.87±0.85) alone. Rapamycin also obviously up-regulated the expression of p-Smad2 and p-Smad3 in the lymphocytes.@*CONCLUSIONS@#Rapamycin can promote the differentiation of Treg cells by up-regulating the expression of p-Smad2 and p-smad3 to improve neurological deficits in mice with EAE.
Subject(s)
Animals , Mice , Anti-Inflammatory Agents , Therapeutic Uses , Cell Differentiation , Encephalomyelitis, Autoimmune, Experimental , Drug Therapy , Metabolism , Interferon-gamma , Metabolism , Interleukins , Metabolism , Lymphocytes , Cell Biology , Mice, Inbred C57BL , Sirolimus , Therapeutic Uses , Smad Proteins , Metabolism , T-Lymphocytes, Regulatory , Cell Biology , Transforming Growth Factor beta , Metabolism , Up-RegulationABSTRACT
Objective To investigate the expression pattern of transcription factor Olig 2 in cuprizone-induced mouse model of acute demyelination .Methods C57BL/6 mice were fed with 0.2%cuprizone to induce acute demyelina-tion.Immunofluorescence and qRT-PCR were used, and Olig2, MBP and GFAP were detected in the brain tissues of con-trol group and cuprizone-treated groups for 6 weeks and recovery for 2 weeks.Results Severe demyelination occurred in the corpus callosum following 6-weeks exposure to cuprizone , while remyelination was detected in the white matter after the mice were given diet without cuprizone .In the normal mice , Olig2 was expressed in a low level , while the experessions of Olig2 and GFAP were significantly increased , and Olig2 +/GFAP+cells were detected after demyelination .But the expres-sion of MBP was below the normal level with demyelination .After recovery for 2 weeks, the experession of Olig2 was lower, but the experessions of MBP and GFAP were increased .Conclusions Olig2 may play an important role in the glial differ-entiation from neural progenitor cells into active astrocytes , and in the glial scar formation .
ABSTRACT
@#Objective To investigate the inhibitory effect of Pingliukang (a prescription) medicated serum on the proliferation of cultured rat C6 glioma cells and influence on cell cycle in vitro. Methods MTT colorimetry were performed to measure the levels of the proliferation of rat C6 glioma cells cultured with 2.5%, 5%, 10% and 20% of Pingliukang medicated serum for 24 h, 48 h and 72 h in vitro. The effect of Pingliukang medicated serum on cell cycle were observed by FCM. Results The proliferation of C6 cell was obviously inhibited by Pingliukang medicated serum with dose-effect relationship. The inhibitory effect of 20% of medicated serum was the strongest. When the C6 glioma cells were treated with 10% and 20% medicated serum for 48 h and 72 h, the cells of S period declined. Conclusion The Pingliukang medicated serum can inhibit the proliferation and block cell cycle of cultured C6 glioma cells.