Effect Of QingHaoBieJia Decoction On TNFα And STNFα R During The Inducing Process Of CD34~+Cell-derived DC Of MRD-L Patients

Abstract:Objective:Investigate the effect of Qinghao Biejia Decoction on minimal residual CD34+cell leukemia induced dendritic cells’ differentiation and maturation, as well as the influence on the secretion of tumor necrosis factor and its soluble receptor. Research the treatment of Qinghao Biejia Decoction on minimal residual CD34+cell leukemia induced dendritic cells’biological effects.Methods:Preparation of drug containing serum of rabbit after perfused with high, middle, low dose Qinghao Biejia Decoction. Mix with heparin after collect the bone marrow from acute myeloid leukemia (AML-CR) patients. Separate the bone marrow mononuclear cells(BMNCs) by FICOLL gradient centrifugation method, and purify CD34*cells by the method of CD34+magnetic activated cell sorting. Amplify CD34+cells in6-9days with TPO, SCF, FLt-3, IL-4in vitro, and culture them with containing rabbit serum and cytokines. Divided them into5groups:Qinghao Biejia Decoction of high, medium and low dose group, the blank serum group. Detecte the TNFα、 sTNF α R from the Oth,6th,9th day’s supernatant serum by ELISA kit. And detecte the dendritic cells surface antigen CD80, CD83, CD86, CDla, HLA-DR by flow cytometry.Results:1.DC s morphological observation:under microscope for morphology, they were CD34+cells on Oth days., The cells’surface became irregular after3days, which appearance were fuzzy. They clustered and formated colony. The cells became large and protruded branches after6days, interwoven with other dendritic cells to increase the number of colony. They came basically mature and have typical shapes on Ninth days. The QG, QZ, QD induced DCs were more typical than the KB, YZ group, and the number of DC cells in group QG was moer than QZ, QD group, KB, YZ group showed no significant difference.2. DCs’immunophenotype:each group can be expressed, The CD80did not differ between groups (P>0.05);the containing serum groups were different between KB group and YZ group in the expression of CD83、CD86, CDla and HLA-DR.And the QG group was the best(P<0.05) compared with others.3. Detection of3. TNFα, sTNF α R:the content of TNFα and sTNFR in each group were decreased, and there were significant differences (p<0.05) between the9th day,6th day and Oth day. The QG group, QZ group, QD group were different from others, but there was no different between themselves’.The sTNF α R detection on the9th day was different from6th day and Oth day respectively. There was no significant difference between the Oth and6th day (p>0.05). The QG group had significant difference (p<0.05) between the other4groups. QZ group, QD group and KB group, YZ group had difference(p<0.05), but there wsa no significant difference between the two groups. KB group and YZ group were no significant difference.Conclusion:1.The QingHaoBieJia Decoction could promot the maturation of CD34+Cell-derived DC directly or indirectly through changing the content of cytokines.2. The Medicated Serum combined with cytokines can promot the maturation of CD34+Cell-derived DC, higher expression of CD83, CD86, CDla, HLA-DR.3. QingHaoBieJia Decoction can reduce the secretion of TNF a and sTNF a R, and the high dose group does the best.