圣愈汤及其拆方调控血虚小鼠造血生长因子的研究

【作者】 王均宁；

【导师】 刘持年；

【作者基本信息】 北京中医药大学 ， 中医方剂学， 2004， 博士

【摘要】 血虚证是临床常见证候之一,常见于多种疾病病程中。方剂的配伍理论是方剂学理论的精髓,也是方剂学研究的重点内容之一。论文以中医理论为指导,在系统分析血虚证的实验研究和临床文献资料的基础上,探讨血虚证的发病机制、病势演变规律、治疗思路及其组方用药规律,并通过对圣愈汤调控血虚小鼠造血的作用机制及配伍机理的实验研究,以诠释中医补血方剂的组方配伍机制和可能的作用机理。本文分二部分:第一部分理论研究,通过对血虚证的发病机理和病机演变特点的深入分析,提出了损气、耗津(阴)、致瘀是影响血虚证病势演变的潜在病机,也是影响血虚证病势进退的重要因素和治疗的关键环节。提出了补其不足,资其化源,先其所因,以制其变的组方思路。认为补血剂中配伍补气、滋津(阴)、行血药的意义,不只是为增强补血之功,更重要的还在于针对损气、耗津(阴)、致瘀的潜在病机进行干预,以扭转病势转归的不良演变,促进疾病的向愈转化。对补血名方圣愈汤的源流发展、组方意义进行了深入分析,并对补血剂的组方配伍规律进行了探讨。第二部分实验研究,以 Coγ射线诱发小鼠血虚模型对圣愈汤调控造血的疗效及作用 60机制进行了探讨。实验中主要检测了小鼠外周血液中的有形成分、血清EPO、IL-6 水平、骨髓细胞的GM-CSF 表达以及CD34 细胞比例和细胞周期,观测了骨髓组织的损伤与修复 +情况。结果显示圣愈汤能明显升高血虚小鼠血液中的WBC、RBC、HGB、PLT,提高胸腺及脾脏指数,升高循环中 EPO、IL-6 水平,增强骨髓造血细胞 GM-CSF 蛋白表达和 CD34分子的抗原表达,诱导骨髓细胞由静止期进入增殖周期,抑制细胞凋亡,促进骨髓造血功能的修复。提示本方调控造血的作用机制可能是主要通过调控造血生长因子EPO、IL-6 的分泌和GM-CSF 蛋白表达,促进骨髓造血干/祖细胞的增殖、分化,刺激血细胞的生成和释放来实现的。这也很可能是补血剂发挥“补血”作用的重要机制。拆方研究表明,各拆方组分的作用靶点和作用强度存在一定的差异,地黄、芍药组合升高全血红细胞、血红蛋白、促进EPO 分泌的作用较好,与川芎、当归合用效果更佳,人参、黄芪的加入,在某种程度上似对 EPO 的分泌有抑制作用,可避免因循环中 EPO 水平的过度升高而引起的不良反应;而含人参、黄芪的组方对IL-6 分泌、MG-CSF 表达均有促进作用,加入川芎、当归可使作用明显增强;当归、川芎的加入,能够显著增强熟地、白芍及人参、黄芪、熟地、白芍刺激骨髓造血细胞由相对静止时相进入细胞增殖活跃时相,促进骨髓细胞 DNA 合成<WP=7>2 圣愈汤及其拆方调控血虚小鼠造血生长因子的研究的作用;当归、川芎能增强熟地、白芍、人参、黄芪及人参、黄芪增强熟地、白芍、当归、川芎组合(即四物汤)促进骨髓造血干/祖细胞的增殖作用。本研究结果反映了圣愈汤各药组之间既相济相助、又相制为用的组方配伍特点,以及复方活性多组分、作用多途径的整体优势,对血虚证的治疗过程,突出的是多层次、多脏器的调控作用,符合中医方剂组方及效用特点。在一定程度上反映了益精补血药在补血剂中的主导作用,体现了在补血剂中配伍补气健脾药、滋生阴津药、调血行血药在调控造血的某些环节中的协同增效或相制为用的组方特点,对于促进造血功能尽早恢复、抑制病势的进一步演变发展具有重要意义。从而初步验证了针对血虚证损气、耗津(阴)、致瘀的潜在病机,补血兼以补气、滋津(阴)、行血可扭转病势转归的不良演变的工作假说。初步揭示了益精生血、补气生血及行血活血可助生血的科学内涵。这同时也为该方临床治疗多种原因引起的贫血、防治恶性肿瘤放疗乃至化疗引起的骨髓抑制提供了实验依据。更多还原

【Abstract】 Blood deficiency is a common syndrome of TCM in clinic, which appears in the course ofmany diseases. The compatibility theory is an essence and important component in Formulas ofTCM. Under the guidance of TCM, based on the experimental and clinical literature, we explorethe pathogenesis, development and treatment of blood deficiency. In order to reveal thecompatibility principle and possible mechanism of the nourishing blood formulas, we made anexperiment on the effects of regulating hemopoietic growth factors in blood deficiency mice andthecompatibilitymechanismofShengyuDecoction. Thearticlecontainstwoparts. Part one is theoretical study. We analyze the pathogenesis, development of blood deficiencytheoretically, and consider that damage to Qi, damage to liquid and blood stasis are the potentialpathogenesis and important factors affecting prognosis and treatment of blood deficiency. We alsoconsider that tonifying Qi, nourishing Yin, activating blood not only strengthen enriching blood,but also intervene the potential pathogenesis and restrain further development of the disease. Westudy the origin and development and the compatibility significance of Shengyu Decoction, andexplorethecompatibilityprincipleofthenourishingbloodformulas. Part two is experimental study. This study is to explore the effect and mechanism of ShengyuDecoction on blood deficiency. the Shengyu Decoction was given to mice with blood deficiencyinduced by γ-rays, and then the blood cell, platelet, EPO and IL-6, also GM-CSF, CD34+ andcell cycle in bone marrow cell were studied thoroughly. the damage and repairment of myeloidtissue were observed. The results showed that Shengyu Decoction can increaseWBC, RBC, HGBandPLTinblood.ItseffectonWBCandPLTwassuperiortoFourHerbsDecoctionandtheeffecton RBC and HGB was inferior. The results also showed that EPO, IL-6 in blood serum and theexpression of GM-CSF in bone marrow cell increased. We deduce that the enriching blood effectwas close related to EPO, IL-6, which can induce bone marrow cell from resting phase toproliferating phase, accelerate the proliferation and differentiation of hemopoietic stem cell,stimulate the production and releasing of blood cell. These also may be the most importantmechanismofnourishingblood. Research on the different ingredient groups indicated that the different ingredient groups ofShengyu Decoction had different target and strength. The groups with prepared rhemanniarhizome and white peony root were superior to the groups without them in enhancing the RBC,HGB and EPO, and when prepared rhemannia rhizome and white peony root combined withChinese angelica root and chuanxiong rhizome, this effect can be improved remarkably, andWhen combined with ginseng and astragalus root, EPO in blood was not increased, it is seemed<WP=9>4 圣愈汤及其拆方调控血虚小鼠造血生长因子的研究thatginsengandastragalusrootcaninhibittheexcretionofEPO. In the groups with ginseng and astragalus root IL-6 in blood serum and the expression of theMG-CSF in bone marrow cell increased, when combined with Chinese angelica root andchuanxiong rhizome, this effect was improved. The herbs of Chinese angelica root andchuanxiong rhizome combined with prepared rhemannia rhizome , peony root and ginseng,astragalus root, Chinese angelica root and chuanxiong rhizome can promote hemopoietic cell intoproliferating phase from resting phase, and DNA synthesis of bone marrow cell. When Chineseangelica root and chuanxiong rhizome combined with the groups containing prepared rhemanniarhizome, white peony root, ginseng and astragalus root, the effects of improving the proliferationand the differentiation of HSC can be enhanced. The similar effects also observed in the groupwithGinseng,astragalusrootcombinedFourHerbsDecoction. Characteristics of compatibility principle of Shengyu decoction are that the ingredientsinteract with each other cooperatively or restrictedly. multiple acti更多还原