| BackgroundCancer is the second reason that causes the human death in the world, but the most important way to treat tumor is radiotherapy and chemotherapy in clinics. As we all know, most of the radiotherapy and chemotherapy have inhibition effects to bone marrow. Leucocytopenia is the most important side effect that will lead to the failure of therapy because of secondary infection. As a result, the death rate will be increased because some better therapy scheme cannot continue. It is the key point to ensure the effect of radiotherapy and chemotherapy to cancer that improving the recovery effect of immunity system. Recent evidence indicates that CD34+ stem cells is the earlist hematopoietic cells with the developing of isolating method of hematopoietic stem cells, which have the capability to differentiate to lineage mature functional cells under the stimulate of inside and outside cell environment and cell factor. Cepharathine Hydrochloride (CH) is a bisbenzy lisoquinoline alkaloid isolated from the tubers of Stephanie delavayi Diels is half synthesized CH has the stronger biological actives such as the effect of eliminate inflammation, suppress germ and regulate immunity. This study confirmed the leukocytotic effect of CH through imitated the inside environment of tumor patients and made the leukocytopenia model, and investigated the relationship between leukocytotic reason and the proliferation of CD34+ stem cells, provided theory foundation for the further development, exploitureand research of CH.MethodsMurine xenografts model of mouse hepatocarcinoma cell line H22 was used in vivo, the pharmacodynamic studies that CH prevent and treat leucocytopenia caused by cyclophosphamide(CTX). Bone marrow cells was isolated and cultured with CH while CTX inhibited the differentiation of stem cells in vitro. The effect that CH proliferated bone marrow CD34+ stem cells was observed. Leucocytopenia model was used to study the relationship between therapy of CH on Leucocytopenia induced and bone marrow cells (BMC) and bone marrow CD34+ hematopoietic stem cells in mice. Adjuvant antitumor effect of CH in vivo was investigated in a xenograft animal modle.Results1. The effect of CH on proliferating bone marrow CD34+ hematopoietic stem in vitroThe value of positive percentage rate of CD34 cells: blank, 40ng.ml-1CH, 20ng.ml-1, 10ng.ml-1CH and 50ng.ml-1G-CSF was (49.38±1.12)%,(83.67±2.72)%, (64.94±3.77)%,(53.58±1.72)%,(61.84±2.51)%.Comparing with CTX control groups's CD34-positive values with every concentration of CH and G-CSF, there were significant differences (P<0.05) . It also has statistically significant differences (P<0.05) between 40ng.ml-1 CH groups' CD34-positive value and the CTX control group's CD34-positive value.2. Pharmacodynamic Studies2.1 The precaution effect of CH on leucocytopenia caused by chemotherapy:The WBC of tumor-bearing group, 30mg.kg-1CTX group, 30mg.kg-1+20mg.kg-1CH and 30mg.kg-1+10mg.kg-1CH groups is (10.93±1.34)×l09, (1.83±0.31)×l09,(4.76±0.56)×l09, (4.03±0.58)×l09, the WBC number of CTX was the lowest. CHgroups compared with CTX control group, there were significant differences(P<0.05) . It is 6 days after stop drugs, the WBC has recovered, each groups was(9.79±1.76)×l09, (3.88±0.59)×l09, (9.83±1.24)×l09, (8.99±1.30)×l09. There were statistically significant differences (P<0.05) between experimental groups' WBC values and the CTX control group's WBC value.2.2 The therapy effect of CH on leucocytopenia caused by chemotherapyThe WBC of G-CSF was recovered in third days after beginning give drugs, the number of Tumor-bearing group, 30mg.kg-1CTX group, 20mg.kg-1 CH group, 10mg. kg-1 CH group and 50μg.kg-1 G-CSF group is:(9.80±1.50)×109, (2.35±0.51)×109, (5.48±0.88)×109,(4.38±0.67)×109, (10.64±2.70)×109 and The peak of WBC was higher than other days in fifth days, the value is:(9.48±1.72)×109, (3.98±0.54)×109, (9.49±1.00)×109,(7.28±0.47)×109, (19.48±1.75)×109,we can analyse from the date, the leukocyte of 20mg.kg-1CH has recovered. Both of CH groups have recovered in sixth day, the value of every groups was:(9.61±1.46)×109, (4.91±0.63)×109, (10.01±1.30)×109, (9.09±1.42)×109, (17.06±1.35)×109.There were statistically significant differences (P<0.05) between experimental groups' WBC values and the CTX control group's WBC value.3.The effect of CH on proliferating BMC and bone marrow CD34+ hematopoietic stem cells in vivo3.1 Proliferating BMC and bone marrow CD34+ hematopoietic stem cells in vivo with precautionary experiment. The BMC of bone marrow of each group were Tumor-bearing group (7.79±0.90)×106, 30mg.kg-1CTX group (1.53±0.30)×106, 30mg.kg-1CTX+20mg.kg-1 CH group(4.52±0.59)×106, 30mg.kg-1CTX+10mg.kg-1 CH group (3.63±0.54)×106,respectively,If the BMC of different concentration CH group was compared with that of the CTX group, there was statistical significant difference (P<0.05).The result of flow cytometry indicated that every group were (4.99±1.38)%, (1.67±0.72)%, (20.63±4.36)%, (11.47±2.59)%. There was statistical significant difference (P<0.05) between CH control group's CD34+ values with that of CTX group's.3.2 Proliferating BMC and bone marrow CD34+ hematopoietic stem cells in vivo with therapeutic administrationThe result of the BMC were Tumor-bearing group (7.61±1.25)×106, 30mg.kg-1CTX group (1.59±0.37)×106,20mg.kg-1 CH group (4.92±0.68)×106, 10mg.kg-1 CH group (3.63±0.69)×106, 50μg.kg-1 G-CSF group (7.10±0.66)×106 and the result of CD34+ BM cells were (5.40±1.22)%, (1.66±0.38)%, (58.66±8.19)%, (37.39±3.95)%, (18.00±4.12)%. Both BMC and CD34+ BM cells were statistical significant difference (P<0.05) compared CH and G-CSF control group's with CTX group's.4. Effect of CH on the growth of H22 tumor xeografts in miceThe tumor weights of the Tumor-bearing group CTX group 20mg.kg-1 CH group 10mg.kg-1 CH group and 50μg.kg-1G-CSF group were 2.1 l±0.29g, 1.20±0.20g, 0.93±0.14g, 0.79±0.14g, 1.10±0.26g. The inhibitory rate of tumors of each group except the blank control group were 43.1%, 55.9%,62.6%,47.9. If the tumor weights of the 20mg.kg-1 CH group and 10mg.kg-1 CH group were compared with that of the CTX group, there was statistical significant difference (P<0.05) .Conclusion1. Above studies in the precaution and therapy experiment indicates that CH could inhibit the leukocytopenia caused by chemotherapy.2. CH promotes the proliferation of bone marrow CD34+ hematopoietic stem cells in vivo and in vitro. It may be one of the important mechanisms of increasing WBC numbers.3. The application with CH and CTX has a significant increase in chemotherapy. It suggests that CH can induce the toxic of chemotherapy, as well as, supply scientific theory basis for further study and clinical application...
|
PDF Full Text Request