Effects On Differentiation And Functional Maturation Of Rat Bone Marrow-derived Dendritic Cells By Sinomenine

【Objective】 To Investigate the effects Sinomenine on dendritic cells from rat bonemarrow origin (Dendritic cell, DC) and mature in vitro differentiation anddevelopment.And characterized in terms of morphology, phenotype and antigenpresentation, and to stimulate secretion of T cell activation ability.1Observe thesituation of cell morphology use the inverted phase contrast microscope;【Methods】 Bone marrow-derived precursor cells from rat DC;ImDC isolated andinduced magnetic activated cell sorting purification.Divided into a control group andthe low, medium and high doses of different treatment groups Sinomenine. Harvestethe DC after48hours.Detect the expression level of maturity:2Teck the expressionof cell phenotype CD80, RT1B’s by flow cytometry.3EIISA detect IL-12of cytokines；4.DC stimulated mixed lymphocyte reaction detection of T cell activation,【Results】Measured results1.Inverted microscope were used to observe cellappearance: LPS group cell meet the morphological characteristics of mature dendriticcells. The most cell mature in SN L group; SN M group of partial suspension,maturity difference; SN H group most immature.2.Expression of CD80in LPS group, SN L group, SN M group, SN H groupwere93.54%,79.29%,61.40%,35.20%, Compared with the control group,Sinomenine low, middle and high dose group, expression is significantly lower(p<0.05), the difference was statistically significant. Detection of RT1B expression inLPS group, SN L group, SN M group, SN group H rates were compared to97.70%, 91.20%,84.40%,65.90%and the control group, the expression of sinomenine, in highdose group significantly decreased (p<0.05), the difference was statisticallysignificant, but low dose group showed no significant difference (P>0.05).3. ELISA detection of cytokines in IL-12p70results, the control group and thelow, middle, high dose group were2320.26±60.21,2123.35±64.14850.01±71.21230.19±28.87. After statistical analysis, compared with the control group, AotoNaka, treatment of high dose of DC content of IL-12p70in the culture mediumdecreased (p<0.05), the difference has statistical significance, the content ofsinomenine treatment by a low dose of IL-12p70in the culture medium had nosignificant difference (P>0.05).4. Indicates that the detection of DC mixed lymphocyte reaction to stimulateallogeneic T lymphocyte activation：1:5mixing control, low, medium and high dosegroups OD values were4.53±0.63,4.48±0.50,3.91±0.48,2.85±0.51;1:10mixingcontrol, low, medium and high dose groups OD values were3.86±0.49,3.80±0.41,3.29±0.38,2.61±0.51;1:20mixing control, low, medium and high dose groupsOD values were3.08±0.31,2.99±0.40,2.16±0.42,1.65±0.38;1:40mixingcontrol, low, medium and high dose groups OD values were1.99±0.30,1.93±0.28,1.58±0.34,1.15±0.36.After statistical analysis,compared with the control group,the different mixing proportion, Aoto Naka, high dose group was decreasedsignificantly ((p<0.05), the difference was statistically significant, but no significantdifference between the low dosage group was changed (P>0.05).【Conclusions】A variety of cytokines can induce rat bone marrow derived dendriticcells and promote its maturity, and sinomenine could inhibit DC further mature; lowdose inhibitory effect is weak, with the increase of the dose of sinomenineon inhibition of DC maturation enhancement.