| ObjectiveTumor threaten human health seriously,and traditional medication is under the treatment with severe side effects.Under this condition the current work is searching for highly effective and low-toxicity drugs.Periplaneta americana extract CⅡ-3,developed by yunnan Key Laboratory for Biomedical Research and Development of Insects at Dali University,is mainly peptide-based substances.CⅡ-3 anti-tumor function has been confirmed in many human tumor cells,and its anti-tumor mechanism is not yet totally explained.We have established MFC tumor-bearing mice model and dealt with CⅡ-3 and HFDT1.In order to provide theory and data support for anti-tumor therapy,we study the influence on lymphoid cells of MFC tumor-burdened mice,explore the mechanism of drugs affecting the immune system.MethodsWe selected BALB/c mice with age of 6-8weeks and weight in about 18-20g,and inoculated logarithmic growth of MFC cells in the right armpit of the mice.We established the subcutaneous tumor model in BALB/c mice,and divided them into 5groups including model group,CTX group,the periplaneta americana peptide complex CⅡ-3 high and low dose groups,synthetic peptide HFDT1 group.In addition,Normal BALB/c mice were served as normal control group.The normal control and model group mice were given normal saline by gavage(20mL/kg,qd).The C TX group mice were injected with CTX(45mg/kg,10mL/kg,q2d).The high and low doses of CⅡ-3 groups mice were given with CⅡ-3 by gavage(400mg/kg and 200mg/kg,20mL/kg,qd).The HFDT1 group were injected with synthetic peptide HFDT1(7.8mg/kg,10mL/kg,qd).In addition,the proliferation test in mice need to join Brd U drug for 5 days after giving 5 days’foregoing drugs.All the mices were injected with BrdU(4mg/ml,0.2mL each mice).We obtained blood through the inner canthus vein of mice,and used 70μL blood analysis peripheral blood immune cells.we separated the spleen to make Single cell suspension.FACS analysis the prolifetion of spleen’s immune cells.The NK and CTL cells were separated and pured by immune magnetic beads.CFSE markde the MFC tumor cells.Mixing C TL or NK cell with MFC tumor cells by fixed ratio were incubated for 4h.FACS analysis the killing rate of NK and CTL cell after dyed with PI for 15min.Results1.Compared with normal group and CTX group,The CⅡ-3 group and HFDT1group increased periphetal blood lymphocytes,B cells and NK cells(P<0.05).Compared with normal group,Model group’lymphocyte ratio also increased.CTX group’lymphocyte ratio was obviously lower than normal group and model group(P<0.05).2.Compared with model group and C TX group,the CⅡ-3 group and HFDT1group increased peripheral blood T cells,CD4~+T cells and C D8~+T cells(P<0.05);Compared with normal group,the CⅡ-3 group and HFDT1 group increased peripheral blood T cells and CD8~+T cells(P<0.05).3.Compared with normal group and CTX group,the high dose CⅡ-3 group and HFDT1 group enhanced the proliferation of B cells,NK cells and T cells.the low dose CⅡ-3 group also enhanced the proliferation of NK cells and T cell(P<0.05).4.Compared with normal group and CTX group,the low dose CⅡ-3 group enhanced the proliferation of CD4~+T cells and CD8~+T cells,the high dose CⅡ-3group also enhanced the of proliferation CD4~+T cells(P<0.05);Compared with model group,the low dose CⅡ-3 group enhanced the proliferation of CD4~+T cells(P<0.05).5.Compared with normal group and model group,the CⅡ-3 group and HFDT1group enhanced the function of CTL cells killing MFC tumor cells(P<0.05),when the ratio of CTL effector cel s and MFC target cel s were 5:1.6.Compared with normal group and model group,the CⅡ-3 group and HFDT1group enhanced the function of NK cells killing MFC tumor cells(P<0.05),when the ratio of CTL effector cells and MFC target cells were 5:1.Compared with the CTX group,the high dose group and HFDT1 group enhanced the function of NK cells killing MFC tumor cells(P<0.05),when the ratio of NK effector cells and MFC target cells were 5:1.Compared with model group,the CⅡ-3 group and HFDT1group increased the function of NK cells killing MFC tumor cells(P<0.05),when the ratio of NK effector cel s and MFC target cel s were 20:1.Conclusions1.Periplaneta americana extract CⅡ-3 and synthetic peptide HFDT1 can improve the the proportion of total lymphocytes,B cells,T cells,CD4~+T and CD8~+T cells in the peripheral blood.2.Periplaneta americana extract CⅡ-3 and synthetic peptide HFDT1 can enhance the proliferation of T cells,B cel s and NK cells.3 Periplaneta americana extract CⅡ-3 and synthetic peptide HFDT1 can enhance the function of CTL and NK cells killing tumor cells. |
