Study On Compound Zhebei Granules Reversing Multidrug Resistance In Mice Acute Lymphoblastic Leukemia

Acute lymphocytic leukemia(ALL)is a common malignant tumor of the hematopoietic system.It is characterized by clonal proliferation of abnormally differentiated malignant cells infiltrating the bone marrow,blood and other tissues.It is one of the most common childhood malignant tumors in the world,accounting for about 20%of adult leukemia.Chemotherapy,hematopoietic stem cell transplantation and targeted therapy are the main treatment methods.The current chemotherapy makes nearly 80%of patients have a remission period of more than 10 years.With the occurrence of recurrence,the final cure rate is about 25-40%.Bone marrow transplantation and the application of multi-drug combined intensive treatment tegimes have further improved the clinical cure rate of ALL,but there are still some patients with poor treatment results.Studies have shown that the resistance of leukemia cells to chemotherapy drugs is an important factor for the relapse and refractory ALL.Therefore,it is found that drugs that reverse ALL multi-drug resistance are of great significance to increase the sensitivity of chemotherapeutic drugs and improve clinical efficacy.Objectives:1.To Build the L1210/CDDP mouse model,and observe the effect of Compound Zhebei Granules on ALL from the perspective of pharmacodynamics and multidrug resistance mechanism.2.To construct the L1210/CDDP tail vein model,and explore the therapeutic effect of Compound Zhebei Granules on the tail vein model mice from the perspective of pharmacodynamics.Methods:1.L1210/CDDP cells were cultured by maintaining drug resistance,and 1×106 cells were injected into the right armpits of mice to replicate the tumor-bearing ALL model.The therapeutic effect of Compound Zhebei granules on the model mice was observed.2.Real-time PCR method was used to observe the effect of Compound Zhebei Granules on Mdr-1 gene expression in tumor-bearing ALL mice;immunohistochemical method was used to observe the effect on P-gp,GST,Topo II,Bcl-2 and Bax expression.3.L1210/CDDP tail vein mice model was constructed and used to observe the therapeutic effect of Compound Zhebei Granules.Results:1.L1210/CDDP model was successfully replicated 7 days after cell injection.Treatment with Compound Zhebei Granules combined with cisplatin resulted in reduction of tumor size and mass,increase in tumor inhibition rate and prolonged survival.2.Compared to model group,CDDP group showed increased expression of Mdr-1,P-gp,Topo ? and Bax,and reduced expression of GST and Bcl-2(P<0.05).Compared to CDDP group,medium-dosed Compound Zhebei Granules group and medium-dose combination group showed increased Mdr-1 expression;medium-dose Compound Zhebei Granules group,low-,medium-and high-dose combination groups showed increased P-gp expression;high-dose combination group showed increased Topo ? expression;medium-dose Compound Zhebei Granules group and low-dose combination group showed increased GST expression while medium-and high-dose combination groups showed reduced GST expression;medium-dose Compound Zhebei Granules group showed increased Bcl-2 expression while haigh-dose combination group showed reduced Bcl-2 expression;medium-dose combination group showed increased Bac expression(P<0.05).3.21 days after tail vein injection with L1210/CDDP cells,bone marrow of 6 randomly selected mice were obtained and the blast count was observed to be between 5%to 15%.No blast cell was found in peripheral blood.Compared to control group,the model group showed lower white blood cell and platelet counts(P<0.05).Compared to model group,low-dose combination group showed prolonged survival and lower bone marrow blast count(P<0.05).Compared to CDDP group,low-dose combination group showed higher hemoglobin concentration,white blood cell and platelet counts(P<0.05).Conclusions:Compound Zhebei granules combined with cisplatin can prolong the survival time of L1210/CDDP tumor-bearing mice,reduce tumor mass and volume,and increase tumor inhibition rate.For L1210/CDDP tail vein model,it can prolong the life span of mice,reduce the proportion of bone marrow blast cells,and maintain normal blood parameters,thus playing a therapeutic role.It can improve the therapeutic effect in the two ALL drug resistance models and reverse L1210/CDDP multi-drug resistance through Mdr-1,P-gp,GST,Bcl-2,and Bax,multiple mechanisms enhancing the effectiveness of chemotherapy.