| 【Objective】To investigate the effects of Oxymatrine on maturation and functions of murine bone marrow derived dendritic cells in vitro.【Method】Murine myeloid dendritic cells(DC) were generated from bone marrow in vitro using rmGM-CSF and IL-4, DC were matured by LPS for 48h.Bone marrow derived dendritic cells were treated with 0.5mg/ml Oxymatrine at day 0 and 5,respectively.The control was treatd with medium alone.The phenotypes and functions of DCs were determined at day 7.There were 7 groups in this experiment: group A: DC; group B: DC+OMT (D0); group C: DC+OMT (D5); group D: DC+LPS; group E: DC+OMT(D0)+LPS; group F: DC+OMT(D5)+LPS; group G: T cell control.The phenotype of DC was determined by flow cytometry. The inhibition of dendritic cells and the effect of Oxymatrine on immunno-stimulatory capability of DCs was determined by【3H】Thymidine incorporation assay.ELISA was used to measure the consentration of interferon-γ(IFN-γ) in supernatants of cell cultures.【Result】1. The proliferation of dendritic cells was inhibited by 0.8mg/ml oxymatrine.Ranging from 1.0mg/ml to 5.0mg/ml, oxymatrine had the dose - dependent inhibitory effect on the proliferation of dendritic cells, which showed significant difference compared with control group (P 0.05). 2.The expression of CD40 on DCs:①The expression of CD40 on DCs treated with Oxymatrine at day 0 which was group B was higher than the control group A【n=4, (40.77±1.28)% vs (25.93±7.85)% (p<0.001)】;the expression of CD40 on DCs between group C and A had no significantly difference.②The expression of CD40 on DCs treated with Oxymatrine at day 0 and induced further maturation by LPS,as we named group E was higher than the control group D【n=4,(48.62±2.46)% vs (34.92±5.82)% (p<0.05)】.3.Mixed lymphocyte reaction:①The cellular proliferation of splenocytes stimulated by DCs were higher than the T cell control which was group G(p<0.05), whatever the ratioes between DCs and T cells were 1:5, 1: 10 and 1:20.②When DCs and T cells were in the ratio of 1:5,the cellular proliferation of splenpcytes stimulated by group B DCs was higher than the control group A【n=3, (49094.67±3048.50)cpm vs (26628.67±4268.55)cpm (p<0.05)】.There was no signficant difference between group C and A.③As DCs were induced further maturation by LPS, the cellular proliferation of splenpcytes stimulated by group E DCs was higher than the control group D【n=3, (149144.30±7212.92)cpm vs (120832.30±6825.45)cpm (p<0.05)】.4. We detected the concentration of INF-γin supernatants of MLR,when DCs and T cells were in the ratio of 1:5.①The concentration of INF-γin MLR supernatants of group A-F were higher than the T cell control (p<0.05).②The concentration of INF-γin MLR supematants of group B was higher than the control group A【n=3, (181.84±33.16)pg/ml vs (100.88±8.51)pg/ml (p<0.05)】. There was no signficant difference between group C and A.③As DCs were induced further maturation by LPS, The concentration of INF-γin MLR supematants of group E and F were not higher than the control group A.There were no signigficant difference among these three groups..【Conclusion】Oxymatrine can increasing the expression of CD40 on the dendritic cells and the capacity of stimulating lymphocyte proliferation and excretion ofγ-IFN.It can strengthen the dendritic cells functions of presenting antigen.It can promote the mature of dendritic cells induced by LPS. The specific mechanism need deeper study. |
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