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The Relationship between Therapy of Cepharathine Hydrochloride on Leucocytopenia Induced by Chemotherapy and Bone Marrow CD34～+ Hematopoietic Stem Cells in Mice

Author: LiJie
Tutor: WangQingDuan
School: Zhengzhou University
Course: Pharmacology
Keywords: Of Stephania base hydrochloride Leukocyte CD34 ~ stem cells Nucleated bone marrow cells
CLC: R730.5
Type: Master's thesis
Year: 2007
Downloads: 104
Quote: 0
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Abstract


The background cancer is the second leading cause of the world the human toll, the clinical treatment of tumors is still the important means of radiotherapy and chemotherapy. As we all know, the majority of radiotherapy and chemotherapy on bone marrow has a certain inhibition therapy often cause severe peripheral leukocytes reduced, resulting in immune function and reduce the patient secondary infection, and eventually lead to treatment failure. Thus, the prevention and treatment of bone marrow suppression, speed up the discharge, the reconstruction of the immune system after chemotherapy, guarantee malignancy put the key of the effect of chemotherapy. As hematopoietic stem cell separation and purification technology progress the of CD34 hematopoietic stem cell research early hematopoietic cells, it can, in the role of cytokines in the stimulation of intracellular and extracellular environment to each department as functional differentiation and maturation of cells. The Stephania alkali (cepharathine) is a two-benzyl isoquinoline alkaloids from Menispermaceae Stephania is a plant extract separated by salt to Stephania hydrochloride the alkali (Cepharathine Hydrochlorid CH), which has the role of anti-inflammatory, antibacterial, regulation of the immune function, this study simulated clinical tumor patients in vivo environment in the body, resulting in leukopenia model, confirmed hydrochloric Stephania alkali effect of elevated white blood cell, and explore a liter in both in vitro and in vivo leukocyte the the mechanism promote bone marrow of CD34 hematopoietic stem cell proliferation relationship, provides a theoretical basis for further research and development for the of Stephania base hydrochloride. The method experimental animals transplanted tumor hepatoma H22 cells transplanted subcutaneously model, pharmacodynamics observed leukopenia caused by the different concentrations of CH prevention and treatment of cyclophosphamide. In vitro experiments using isolated and purified nucleated bone marrow cells were cultured with different concentrations of CH inhibit the differentiation of stem cells, determination of CH on bone marrow CD34 hematopoietic stem cell proliferation in vitro; vivo experiments cyclophosphamide (cyclophosph-amide, CTX) caused by Dutch H 22 -cell mouse leukopenia model, observe different concentrations of CH prevention and treatment of leukopenia induced by CTX mouse bone marrow nucleated cells (Bone Marrow Cells BMC) and CD34 hematopoietic stem cell proliferation and tumor growth. Results 1. Impact of different doses of hematopoietic stem cell proliferation in vitro of bone marrow CD34 CH nucleated bone marrow cells co-cultured for 5 days, the results of the control group (49.38 ± 1.12)%, 40ng/mlCH (83.67 ± 2.72) %, 20ng/ml CH (64.94 ± 3.77)% 10ng/ml CH group (53.58 ± 1.72)%, 50ng/ml G-CSF group (61.84 ± 2.51)% administered group and negative control group the difference was statistically significant (P <0.05), CH high-dose group compared with the G-CSF group difference was statistically significant (P <0.05). 2. Induced leukopenia prevention role to chemotherapeutic drugs in vivo leukocyte 2.1: The results found that 24 hours after stopping white blood cell count were: tumor-bearing control group (10.93 ± 1.34) × 10 9 , 30mg/kgCTX group (1.83 ± 0.31) the × 10 , 30mg/kgCTX 20mg/kgCH group (4.76 ± 0.56) are × 10 9 , 30mg/kgCTX 10mg/kgCH group (4.03 ± 0.58) × 10 9, CTX group of white blood cells to a minimum. Different concentrations of CH group of white blood cell count the CTX group the difference was statistically significant (P <0.05). After stopping 6d administration group, white blood cells are returned to normal white blood cell count in each group were (9.79 ± 1.76) are × 10 9 (3.88 ± 0.59) are × 10 9 , (9.83 ± 1.24) are × 10 9 (8.99 ± 1.30) × 10 9 dose group of white blood cell count compared with the CTX group differences were statistically significant (P <0.05). 2.2 to chemotherapy induced leukopenia therapeutic effect: different drugs treated white blood cell count results found 50μg/kgG-CSF group of white blood cells in the administration 3d back to normal white blood cell count, each dose group were: tumor group (9.80 ± 1.50) × 10 9 , 30mg/kgCTX group (2.35 ± 0.51) are × 10 9 20mg/kgCH group (5.48 ± 0.88) × 10 9 , 10mg/kgCH group (4.38 ± 0.67) × 10 9 , 50μg/kgG-CSF group (10.64 ± 2.70) are × 10 9 5d reached peak order (9.48 ± 1.72) × 10 9 (3.98 ± 0.54) are × 10 9 (9.49 ± 1.00) × 10 , (7.28 ± 0.47) × 10 9 , (19.48 ± 1.75) × 10 9 , can be seen from the data, CH high-dose group of white blood cells has been restored properly. Each dose group after administration 6d all back to normal, are as follows: (9.61 ± 1.46) the × 10 (4.91 ± 0.63) are × 10 9 , ( 10.01 ± 1.30) × 10 9 , (9.09 ± 1.42) × 10 9 , (17.06 ± 1.35) × 10 9 , each administration group difference was statistically significant (P <0.05) compared with the CTX model group. 3. In vivo bone marrow hematopoietic stem cell proliferation in the number of nucleated cells and CD34 3.1 prophylaxis in mouse bone marrow nucleated cells and CD34 hematopoietic stem cell proliferation: drugs treated bone marrow nucleated cell count results were: tumor-bearing control group (7.79 ± 0.90) × 10 6 30mg/kg CTX group (1.53 ± 0.30) × 10 6 < / sup>, 30mg/kgCTX 20mg/kgCH group (4.52 ± 0.59) × 10 6 30mg/kgCTX 10mg/kg CH group (3.63 ± 0.54) × 10 6 , the treatment group compared with the CTX group differences were statistically significant (P <0.05); flow cytometry CD34 cell expression of bone marrow cell suspension, each set of results: (4.99 ± 1.38)%, (1.67 ± 0.72)%, (20.63 ± 4.36)%, (11.47 ± 2.59)%. Treatment group compared with the CTX group difference was statistically significant (P <0.05). 3.2 Therapeutic use of bone marrow nucleated cells and CD34 hematopoietic stem cell proliferation: After the administration of bone marrow nucleated cell count were as follows: control group (7.61 ± 1.25) × 10 6 , 30mg/kg CTX group (1.59 ± 0.37) × 10 6 , 20mg/kgCH group (4.92 ± 0.68) × 10 6 10mg/kg CH group (3.63 ± 0.69) x 10 6 , 50μg/kgG-CSF group (7.10 ± 0.66) × 10 . Hematopoietic stem cells positive rate of CD34 , respectively: (5.40 ± 1.22)% and (1.66 ± 0.38)%, (58.66 ± 8.19)%, (37.39 ± 3.95)%, (18.00 ± 4.12)%, treatment group compared with the CTX group difference was statistically significant (P <0.05). 4 role in the treatment of drug adjuvant chemotherapy: tumor weighing results: the control group (2.11 ± 0.29) g 30mg/kgCTX group (1.20 ± 0.20) g 20mg/kgCH group (0.93 ± 0.14) g, 10mg/kg CH group (0.79 ± 0.14) g, 50μg/kg G-CSF group (1.10 ± 0.26) g, In addition to the tumor-bearing group remaining groups inhibition rates were 43.1%, 55.9%, 62.6%, 47.9%. The difference was statistically significant (P <0.05) compared with the CH group and CTX group. Conclusion 1. CH has a significant cancer prevention and treatment of chemotherapy-induced leukopenia role. 2. CH in vivo and in vitro can promote bone marrow the of CD34 hematopoietic stem cell proliferation, which may be one of the important mechanisms for CH elevated white blood cell. 3. CH application has obvious synergies with chemotherapy drugs. The prompts CH has a role to reduce the toxicity of chemotherapy to enhance the efficacy of chemotherapy. To provide for the future of CH-depth study and clinical application of scientific theory.

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