Studies On The Effects Of Qi-boosting Toxin-resolving Formula On The Regulatory T Cell Differentiation Induction Effect Mechanism

Objective:To investigate the effect of Qi-Boosting Toxin-Resolving Formula(QBTRF)on the differentiation inducing effect on T cells in the implanted tumors with human nasopharyngeal carcinoma(NPC)cell line CNE2 among BALB/c nude mice and the regulatory effect on the mechanisms responsible for intervening effects of herbal medicines with the effects of Qi-boosting toxin-resolving in the tumor microenviroment of NPC focus.Methods:Nude mice were used in this study to prepare implanted tumor models with human NPC cell line CNE2 by injecting method subcutaneously.Thirty two modeling mice were randomly divided into four groups,i.e.modeling group(MG),Integrative Medicine Group(IMG)with QBTRF and DDP,treating group(TG)treated with QBTRF and positive control group(PCG)treated by DDP.Then,they were treated with various treating procedures,with normal saline for the animals in MG,concentrated solution of QBTRF orally taken plus intraperitonealy injected DDP to those in IMG,concentrated solution of QBTRF given to those in TG,with concentrated solution of QBTRF given by forcedly oral perfusion,and DDP injected intraperitoneally to those in PCG respectively.Followed was observation on the dynamically growing status of implanted tumors during the experimental period among all groups,with growth curve drown by measuring the volumes of implanted tumors and body weight of animals every day for mice in each group.By the end of experimental period,all mice were put into death to dissect tumors for measuring the weight of implanted tumors to calculate the rate of implanted tumor growing-inhibiting and to take samples from the dissected tumors for histopathological observation on the cellular adhesive structure,by light microscopy respectively.Meanwhile,immunohistochemistry was made to detect the expressive levels of CD4,CD8 and Foxp3 in tumor tissues from all groups of animals.These results were also further confirmed by the procedures of Western blots.The RT-PCR technology was used to detect the expressions levels of all specimen.Results:1.As measured on the volumes of implanted tumors after treatment with a comparative analysis with that of MG,there were statistically significance in the differences of tumor volume among the nude mice of IMG,TG and PCG(P<0.01-0.05),with the inhibitory rates of tumor growing being 58.40%,18.70%and 43.80%for them respectively.2.Transplanted tumor tissues prepared for light microscopy of model group showed that their nucleolus was located in the central nucleus nucleolus.For light microscopy,the tissue samples were embedded in paraffin to make series of sections respectively,stained by conventional methods of hematoxylin-eosin procedure,for comparative observation on the histopathological changes among different groups.3.As determined by SABC imunohistochemistry followed by image analysis on the various groups,the expressive activities of CD8 were inhibition in the samples of MG than that of IG,TG and PCG,compared with CD8,CD4 and Foxp3 were much higher in the samples.4.The results of RT-PCR experiment:in the control group of IG,TG,PCG and MG TGF-? mRNA expression amount order 0.469±0.047?0.351±0.063,0.301±0.057,0.833±0.045,Foxp3mRNA expression of 0.569±0.037,0.512±0.036,0.339±0.058,0.306±0.062andIL-10mRNA expression of 0.896±0.052,0.803±0.080,0.412±0.026,0.409±0.029,with significantly statistical differences among them P<0.05.Conclusion:QBTRF inhibiting growth in nude mice and the mechanism may be strengthening its own secretion of cytokines and activation of effector T cell efficiency,mainly through down-regulation of the corresponding tissue CD4,Foxp3 expression,the up-regulation of CD8,IL-10 and TGF-? expression which reduces immune tolerance phenomenon.