ABSTRACT

ABSTRACT

Objective: To evaluate the impact of Peishishengxue Granule (PG) on immune function of H₂₂-bearing mice and explore the mechanism through observing the effects of PG on the immune organs (the thymus, the spleen), the lymphocyte proliferation, and the secretion of the cytokines (IL-2, IFN-γ).

Methods: The models of solid tumor were developed by hepatoma cell H₂₂ transplantation in mice. When the experiment finished, the weights of thymus and spleen were measured. The thymus index (TI) and the spleen index (SI) were calculated. The proliferation of lymphocyte was assessed by MTT assay. The concentration of interleukin-2 (IL-2) in splenocyte culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA), and the mRNA levels of the cytokines (IL-2, IFN-γ) in splenocytes were assayed by reverse transcription-polymerase chain reaction (RT-PCR).

Results: The TI and SI was increased in PG groups. In high dose group they were significantly increased (p<0.05), and rose by 36.0% and 17.9% respectively; The TI in medium dose group was 18.0mg/10g, and rose by 29.5% (p<0.05). The proliferation of lymphocyte was higher in PG groups than that in model group, especially it was markedly higher in high dose group (p<0.05). The concentration of IL-2 in splenocyte culture supernatants was increased in PG groups, significantly in high dose group (p<0.05). The mRNA levels of IL-2 and IFN-γ were higher in PG groups than those in model group, markedly higher in medium dose group and high dose group (p<0.05).

Conclusion: PG has the function of weightening the immune organs (the thymus, the spleen), stimulating activation and proliferation of splenocyte, and facilitating the secretion and

裴正学系列方药的研究

gene transcription of cytokines (IL-2, IFN-γ) in H22-bearing mice. It can improve the immune function of tumor-bearing mouse and inhibit the growth of the tumor.

key words: Peishishengxue Granule(PG); immune function; H22; lymphocyte proliferation; IL-2; IFN-γ

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治疗白血病的专方[1]，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[2,3]。该方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，尤其对放、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[6]运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）；从毒副反应看，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]。

裴氏升血颗粒以扶正固本为大法，注重对机体的整体调节，即增强机体的免疫功能，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[10-18]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瘤小鼠脾淋巴细胞培养上清液中IL-2浓度、脾淋巴细胞IL-2mRNA和IFN-γmRNA转录水平的影响，同时观察其对荷瘤小鼠免疫器官重量、脾淋巴细胞增殖活性的影响，探讨裴氏升血颗粒对荷H22瘤小鼠免疫功能的调节作用，旨在寻找其可能的潜在作用靶点，为裴氏升血颗粒临床广泛应用提供科学、微观的依据。

实验研究