In Vitro Study On Dendritic Cells Function Modulated By Evodiamine

To study the effect of evodiamine on function regulation of dendritic cells (DC) and develop a new immunotherapy strategy of anti-inflammation. It has become a tendency to look for new anti-inflammatory drugs in Chinese traditional herbal medicine and to do new explorations and explanations for their pharmacological actions. Evodiamine, the major alkaloidal component isolated from Chinese herbal drug named Wu-Chu-Yu, exhibits various biological effects, and was reported to has the anti-inflammatory activities. DC, well known as the most powerful or professional antigen-presentation cells (APC), plays a key role in initiating and modulating innate and adaptive immune responses. During the process of maturation, DC's function of antigen uptake and presentation,migration,phenotype and secretion of cytokine influence the systemic immune system. however, there is no report about the effect of evodiamine on function of DC up to now. So it is important to study the effect of evodiamine on DC for modulating the immunologic abnormality induced by inflammatory reaction.Methods: immature dendritic cells (iDC) derived from Balb/c mice marrow cell were induced by GM-CSF for ten days in vitro and received corresponding treatment, then they were randomized into control groups (â… ), evoamine group (â…¡), LPS group (â…¢), evodiamine+LPS group (â…£)1. The capacity of murine marrow derived dendritic cell to present to nonadherent splenocytes after treated with evodiamine was detected by mixed lymphocyte reaction (MLR).2. The secretion level of IL-10 and IL-12 in murine marrow derived dendritic cell when cultured with evodiamine were evaluated by ELISA.3. Flow cytometric analysis was used for analysis of surface CD11c,MHCâ…¡,CD40,CD80 and CD86 of murine marrow derived dendritic cell after treated with evodiamine.4. cDNA microarrarys technique was employed to detect the effects of evodiamine on function regulation gene express in murine marrow derived dendritic cell.Results:1. Absorbance of LPS group and Evo group significant higher than control group(P<0.05); Evo+LPS group obvious higher than Evo group (P<0.05)and Evo+LPS group significant higher than LPS group(P<0.05).2. The secretion level of cytokine in iDC befor or after incubated with evodiamine is too low to be detected and it is no obvious change with the secretion levels of IL-10 and IL-12 in mDC after incubated with evodiamine for 24h and 48h compare with before(P>0.05).3. The expression of MHCâ…¡,CD40,CD80 in LPS group significant higher than control group(P<0.01,P<0.05); the expression of MHCâ…¡,CD40,CD80 and CD86 in Evo+LPS group significant higher than Evo group(P<0.01,P<0.05)and the expression of CD80,CD86 in Evo group were obvious downregulated compare with control group(P<0.05).4. there are seven genes up-regulated and two genes dwon-regulated over two-times withâ…¡/â… ; thirty four genes up-regulated and twelve genes dwon-regulated over two-times withâ…¢/â… ; forty six genes up-regulated and seven genes dwon-regulated over two-times withâ…£/â…¡; twenty four genes up-regulated and two genes dwon-regulated over two-times withâ…£/â…¢.Conclusion:1. Evodiamine not only promotes the antigen presentation of the iDC but also enhances the antigen presentation of the mDC.2. Evodiamine has no significant effect on secretion of IL-10,IL-12 in iDC and mDC.3. Evodiamine inhibit the expression of partial costimulatory molecules in iDC and has no obvious effect on MHCâ…¡and other costimulatory molecules in mDC.4. the effects of evodiamine on dendritic cell involve in regulation and expression of several genes, these genes contribute to the functions, differentiation and maturation of the dendritic cell, and provid the potentiality to seek the target of medicine.