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1、 2A B S T R A C T Objective: Bazhen Decoction cured syndrome of deficiency of blood and qi in traditional Chinese medicine of clinic, it was a classics famous prescription of nourish the blood and promoting blood flow. In the present study, our aim is to explore the effect of Bazhen Decoction on hem
2、atopoietic regulation and its mechanisms and provide the experimental data for the improvement form of medication. The experimental research has original significance. Methods: By using experimental hematology techniques including: Hemopoietic progenitor cell culture, immunohistochemistry, flow cyto
3、metry, and reverse transcriptase- polymerase chain reaction (RT- PCR)etc. With anemic mice model resulted from myelosuppression by complexes methods, Effects and mechanisms of extracts from tonifying qi herbs studied systematically on cell, protein and gene levels. Results: within the range of certa
4、in dosage or concentration: (1) Bazhen Decoction could promote obviously the increase of the number of bone marrow cell (BMC), the number of white blood cell (WBC), red blood cell (RBC) and hemoglobin (HB). It is also promote evidently the increase of blood platelet (PLT). (2) Bazhen Decoction could
5、 promote the entry of BMC into cell cycle, and decrease the cell proportion of G0/G1 phase, and increase the cell proportion of S phase and G2/M phase in myelosuppressed mice. It also could increase the proliferation index (PI) of BMC. (3) Bazhen Decoction could promote the Colony forming unit- eryt
6、hroid (CFU- E), colonyformingunit granulocytemacrophage (CFU- GM), colony forming unit- megakaryocyte (CFU- Meg). The proliferate effect of on hemopoietic progenitor cell of the middle- dosage of Bazhen Decoction Prescription Granules was best. (4) Bazhen Decoction could promote significantly the ex
7、pression of Bcl- 2 protein, inhibit the expression of Bax protein of BMC in myelosuppressed mice. (5) Bazhen Decoction could promote obviously the expression of erythropoietin 3EPO mRNA of blood serum in myelosuppressed mice. Conclusions: Within the range of certain dosage or concentration Bazhen De
8、coction could promote the recovery of hematopoietic function in myelosuppressed mice. Bazhen Decoction could promote the entry of BMC into cell cycle, and decrease the cell proportion of G0/G1 phase, and increase the cell proportion of S phase and G2/M phase in myelosuppressed mice. So it could incr
9、ease the proliferation index (PI) of BMC. Promote significantly the expression of Bcl- 2 protein and inhibit the expression of Bax protein of BMC in myelosuppressed mice. Promote obviously the expression of erythropoietin EPOmRNA of blood serum in myelosuppressed mice. KEY WORDS: Myelosuppressed mic
10、e Bazhen Decoction Cell cycle Apoptosis haematopoiesis regulation前 言 随着社会的发展，人们生活方式和生活环境的改变，医疗模式已由单纯的治疗疾病转变为预防、保健、治疗、康复相结合的方式，传统医学因而再度倍受世人的青睐。尤其是经典方独特的疗效，更是激发了现代医学工作者们的研究兴趣。 八珍汤源于明代薛己的正体类要 ，作为经典方中气血双补的代表方其组成有当归、人参、白术、白茯苓、川芎、白芍药、熟地黄、炙甘草，用于治疗病后虚弱、 各种慢性病， 以及妇女月经不调等属气血两虚者。 中医基础理论认为 “气为血之帅，血为气之母” ， 医方考卷
11、3 记载： “血气俱虚者，此方主之。气者百骸之父，血者百骸之母。气旺则百骸资之以生，血旺则百骸资之以养。 ”现代化学研究1表明,八珍汤富含益于人体的多种微量元素、氨基酸、还原糖、磷脂、维生素、叶酸、甘草酸、芍药苷等活性成分，近几年来八珍汤的研究日趋活跃,随着研究工作的深入开展,其药理研究和临床应用均得到了更大范围的拓展。 在日常生活中，肿瘤仍是人们谈“瘤”色变的一类疾病，放、化疗是肿瘤病人目前常用的治疗手段之一， 但治疗引起的骨髓造血抑制等反应成为影响患者生存质量的重要原因之一，严重者还会危及患者生命。现代医学临床放化疗辅助药物作用较局限，很难较全面的调整由放化疗所致的机体免疫功能损伤，一些药
12、物用量较大且价格昂贵，极大地加重了患者的经济负担。而中医药无疑在这方面具有其独特的优势。 本研究采用临床气血双补的经典方在骨髓抑制小鼠模型上， 应用血液学常规技术、造血祖细胞培养、流式细胞分析术、免疫组织化学、半定量聚合酶链反应等技术，从造血效应，造血祖细胞增殖、细胞周期、凋亡相关蛋白表达、造血细胞因子等多方面探讨八珍汤对骨髓抑制小鼠造血调控的作用及其机制。 为益气补血的中医理论提供实验室依据。 1缩写词表 缩写词 英文全称 中文全称 AP Ammonium persulfate 过硫酸铵 BMC Bone marrow cell 骨髓细胞 BFU- E Burst forming unit-
13、 erythroid 红系爆式集落形成单位，早期红系祖细胞 BM Bone marrow 骨髓 ECM Extracellular matrix 细胞外基质 CFU- E Colony forming unit- erythroid 红系集落形成单位，晚期红系祖细胞 CFU- GM Colony forming unit- granulocyte- macrophage 粒巨噬系集落形成单位，粒巨噬系祖细胞 CFU- Meg Colony forming unit- megakaryocyte 巨核系集落形成单位，巨核系祖细胞 CY Cyclophosphamide 环磷酰胺 DEPC Die
14、thyl pyrocarbonate 二乙基焦磷酰胺 EPO Erythropoietin 促红细胞生成素 EDTA Ethylene diamine tetraacetic acid 乙二胺四乙酸 FCM Flow cytometry 流式细胞术 GAG Glycosaminoglycans 糖胺多糖 G- CSF Granulocyte colony stimulating factor 粒细胞集落刺激因子 G- CSFR G- CSF receptor G- CSF 受体 G0 cell Quiescent cell G0期细胞，静止期细胞 GM- CSF Granulocyte mac
15、rophage colony stimulating factor 粒- 巨噬细胞集落刺激因子 G1 phase Pre- DNA- synthetic phase G1期，脱氧核糖核酸合成前期 G2 phase Post- DNA- synthetic phase G2期，脱氧核糖核酸合成后期 HB Hemoglobin 血红蛋白 HGFs Hemopoietic growth factors 造血生长因子 HM hemopoietic microenvironment 造血微环境 HS Horse serum 马血清 HSCs Hematopoictic stem cells 造血干细胞
16、IL Interleukin 白细胞介素 M phase Mitotio phase M期，分裂期 2LCA leukocyte common antigen 白细胞共同抗原 L- Glu L- Glutamine L- 谷胺酰胺 MDS Myelodysplastic syndrome 骨髓增生异常综合征 2- ME 2- Mercaptoethanol 2- 巯基乙醇 PI Proliferation index 增殖指数 PLT Platelet 血小板 RBC Red blood cell 红细胞 rhEPO Recombinant Human Erythropoietin 重组人促红细胞生成素 rhG- CSF Recombinant Human G- CSF 重组人粒细胞集落刺激因子 rh IL- 3 Recombinant Human interleukin- 3 重组人白细胞介素- 3 rmGM- CSF Recombinant Murine GM- CSF 重组小鼠粒单系集落刺激因子 rmIL-
