Effect Of Jiawei Qinghao Biejia Decoction On IL-6 During CD34~+ Cell-derived DC Induction In AML-CR Patients

Objective:Under the guidance of the traditional Chinese medicine theory,apply the technology of modem serum pharmacology method,cell culture in vitro and vivo,flow cytometry and ELISA etc to observe the level of IL-6 in the process of induction of CD34+ cell-derived DC in AML-CR patients and discuss the relationship between IL-6 and DC.Methods:New Zealand white rabbits were given by gavage with Jiawei Qinghao Biejia Decoction of high,medium and low dose respectively.Collected rabbit blood from the heart on the third day of the experiment and prepare different concentrations of rabbit serum with drug.Then extract the bone marrow from patients who meet our experiment criterion and separate bone marrow mononuclear cells by the method of FICOLL gradient centrifugation.CD34+ cells were purified using CD34+immunomagnetic beads.After the amplification in vitro by Flt-3,IL-3,TPO and SCF cytokines,different concentrations of rabbit drug serum were prepared for induction in vitro accompanied with IFN-a,GM-CSF,IL-4 and TNF-af,including simple cytokine group,normal rabbit serum + cytokine group,high,medium and low-dose Jiawei Qinghao Biejia Decoction serum + cytokine group.On the 0d,6d and 9d of DC induction collected the serum and measured applied ELLISA to mesure IL-6 level.On the 9d detected the surface antigen expressions of CD80,CD83,CD86,CDla and HLA-DR in the dendritic cells by means of flow cytometry.Results:1.Phenotype of DC:The CD80,CD83,CD86,CDla and HLA-DR surface antigens were expressed in all groups.The expressions of CD80 surface antigen showed no difference between all groups(P>0.05);The expressions of CD83,CD86 and HLA-DR in drug serum + cytokine group showed statistical significance compared with normal serum + cytokine group and simple cytokine group(P<0.05).The expressions of CD83,CD86 and HLA-DR showed no significance between drug serum group and cytokine group(P>0.05).The expressions of CD83,CD86 and HLA-DR showed no difference between normal serum + cytokine group and simple cytokine group(P>0.05);CD1a showed statistical significance between medium and low-dose group compared with high-dose group,normal serum + cytokine group and simple cytokine group(P<0.05).But there is no obvious significance between medium-dose group and low-dose group(P>0.05).CD la showed no statistical significance between high-dose group,normal serum + cytokine group and single cytokine group(P>0.05).2.Level of IL-6:The contents of IL-6 were gradually increasing with DC induction maturity time in drug serum + cytokine group,normal serum + cytokine group and simple cytokine group;The contents of IL-6 showed statistical significance in the the corresponding period between drug serum group + cytokine group,normal serum + cytokine group and simple cytokine group(P<0.05),but showed no statistical significance between high-dose drug serum group and medium-dose group(P>0.05),between low-dose groups(P<0.05).The contents of IL-6 showed no statistical significance between high-dose group and low-dose group(P<0.05).The contents of IL-6 showed no statistical significance between normal serum + cytokine group and simple cytokine group(P>0.05).Conclusions:1.Jiawei Qinghao Biejia Decoction could promote the maturity of CD34+cell-derived DC in AML-CR patients.2.Jiawei Qinghao Biejia Decoction could promote DC to secrete IL-6 in AML-CR patients in the process of CD34+ cell-derived DC maturity.