ABSTRACT

ABSTRACT

Objective: To evaluate the impact of Peishishengxue Granule (PG) on anti-tumor effect and mechanism through observing PG of H22-bearing mouse apoptosis of tumor cells and the expression of NF-κB.

Methods: The models of solid tumor were developed by hepatoma cell H22 transplantation in

裴正学系列方药的研究

mice. When the experiment finished, the weights of thymus or spleen were measured. The thymus index (TI) and the spleen index (SI) were calculated. Tumor-inhibiting-rate was calculated by the tumor weight in mice. NF-κB, cell cycle and apoptotic rate were determined by Flow cytometer.

Results: Animal experiments show that ① The TI and SI were increased in PG groups. In high dose group they were significantly increased (p<0.05), and rose by 36.9% and 19.3% respectively. The TI in medium dose group was 18.1mg/10g, and rose by 31.2% (p<0.05). ② The inhibition rate of the tumor was higher in PG groups than that in model group, especially it was markedly higher in medium dose group (p<0.01). ③ The rate of the expression of NF-κB in medium PG dose group was 15.74%. Compared with model group, there was significant difference (p<0.01). ④ The proportion of G₀/G₁ phase cell in model group was the lowest, whereas the proportion of S phase cell was the highest. The proportion of G₀/G₁ phase cell rose and the proportion of S phase cell fell after treatment. ⑤ The apoptosis rate of the tumor cells was higher in PG groups than those in model group, especially it was markedly higher in medium dose group (p<0.01).

Conclusion: PG plays an important role in anti-tumor effect, especially in medium PG dose group. The mechanisms maybe associated with several aspects. It can play a positive role to improve immunity. It may exert influence on the transcripts of the Gene associated with proliferation and apoptosis of tumor cell. It has the effect of disturbing cell cycle. It can improve apoptosis of tumor cell.

Keywords: Peishishengxue Granule, NF-κB, cell cycle, Apoptotic rate, Inhibition rate, Flow cytometer

前言

裴氏升血颗粒是我国著名中西医结合专家、导师裴正学教授拟定的治疗白血病的专方，主要由六味地黄汤合生脉散加太子参、北沙参、党参等组成，重用太子参、党参、北沙参以健脾；六味地黄汤以补肾。该方三十年前因完全彻底治愈了急性单核细胞性白血病（M₅）患者马长生，1974年在苏州召开的血液病会议上被定名为"兰州方"（2003年，裴氏升血颗粒成功治愈了又一例白血病患者（L₂）刘力刚，先后在《南昌日报》及《甘肃日报》进行报道），1997年该方作为甘肃省医学科学研究院院内制剂，做成"裴氏升血颗粒"。该药剂于2002年起作为甘肃省医科院临床科研课题进行实验研究，通过了成果鉴定。曾获得2003年甘肃省皇甫谧科技成果一等奖。

30多年的临床应用，证明裴氏升血颗粒（即"兰州方"）不仅对白血病有很好的疗效，对恶性肿瘤，以及放、化疗引起的免疫功能低下状态有很好的改善作用。临床实践证明该方对食道癌、肝癌、恶性淋巴瘤、再生障碍性贫血等病有显著疗效，其中完全治愈食道癌2例、恶性淋巴

裴正学系列方药的研究

瘤3例、再生障碍性贫血3例、小肝癌2例[4]。薛氏[5]运用"兰州方"治疗原发性肝癌，使患者肿块明显缩小，肝功能完全恢复，提高了患者的生活质量，延长了患者的生存期；李氏等用兰州方配合化疗治疗癌症10例，疗效观察显示：近期疗效兰州方组优于单纯化疗组（p<0.05）；从毒副反应观察，兰州方组明显轻于单纯化疗组（p<0.01）。裴氏升血颗粒与核素并用治疗肝癌15例，治疗组用核素加服PG，对照组仅用核素，发现治疗组毒副作用明显轻于对照组，二者之疗效对比亦有显著性差异[7]。

近年来通过动物实验研究证明：裴氏升血颗粒可明显升高再障模型小鼠（经3.0GY直线加速器全身照射所建立）的外周血红蛋白、白细胞、血小板值，能明显恢复再障模型小鼠骨髓的造血功能[8]。小鼠脾脏病理组织切片观察显示，裴氏升血颗粒可减轻再障模型小鼠脾脏病理学改变，降低淋巴细胞的凋亡率，促进生发中心的恢复，使白髓面积扩大，被膜变薄，扩张的血窦基本恢复正常。免疫组化法检测脾脏CD4、CD8含量结果表明，裴氏升血颗粒可使再障模型小鼠脾脏CD4含量呈增高趋势、CD8含量呈降低趋势，增高CD4/CD8值，从而提高再障模型小鼠的免疫功能[9]。用MTT比色法测定淋巴细胞增殖，其结果显示淋巴细胞增殖活性增高；用双抗体夹心ELISA法测定脾淋巴细胞培养上清液中IL-2的浓度呈增高趋势；用反转录-聚合酶链式反应（RT-PCR）法测定脾淋巴细胞IL-2mRNA和IFN-γmRNA的表达水平亦呈增高趋势[10]。

裴氏升血颗粒以扶正固本为大法，注重对机体的整体调节，使机体之免疫低下状态得到不同程度的改善，从而达到临床疗效。中医这一"扶正固本"之法与现代医学之免疫学、分子生物学、基因组学等[11-17]有着异曲同工之处，前人在这方面虽然已做了一些工作，但是还有许多方面需要我们进一步去探讨。

本实验通过建立荷瘤（H₂₂）小鼠模型，借助分子生物学技术观察裴氏升血颗粒对荷H₂₂瘤小鼠肿瘤细胞凋亡及NF-κB表达的影响情况，从分子水平探讨裴氏升血颗粒的抗肿瘤效果。旨在从分子水平探讨裴氏升血颗粒的抗肿瘤效果以及中草药抗肿瘤的机制。为该药在抗肿瘤治疗中诱导细胞凋亡、调节细胞信号转导等方面进一步提供实验依据，为其在临床广泛应用中提供充分的科学理论依据，为将该制剂开发成高效防治肿瘤的新型中药制剂奠定基础。

实验研究