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

Research on Pei Zhengxue's Series of Formulas

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

Preface

Peishishengxue Granule is a specialized formula for treating leukemia formulated by Professor Pei Zhengxue, a renowned expert in integrated traditional Chinese and Western medicine in China [1]. It mainly consists of Liuwei Dihuang Tang combined with Shengmai San, supplemented with Taizishen, Beishashen, Dangshen, and other herbs. Taizishen, Dangshen, and Beishashen are used to strengthen the spleen, while Liuwei Dihuang Tang is used to tonify the kidneys [2,3]. This formula gained fame after completely curing a patient with acute monocytic leukemia (M5), Ma Changsheng, in 1974 at a hematology conference held in Suzhou, where it was officially named the "Lanzhou Formula" (In 2003, Peishishengxue Granule successfully cured another leukemia patient, Liu Ligang, with L2 type, as reported in both the Nanchang Daily and the Gansu Daily). In 1997, the formula was developed into "Peishishengxue Granule" as an internal preparation of the Gansu Provincial Academy of Medical Sciences. Since 2002, this formulation has been studied as a clinical research project at the Gansu Provincial Academy of Medical Sciences and has passed the evaluation of research outcomes. It was awarded the First Prize of the Huangfu Mi Science and Technology Achievement Award in Gansu Province in 2003.

Over more than 30 years of clinical application, Peishishengxue Granule (also known as the "Lanzhou Formula") has proven to be highly effective not only in treating leukemia but also in improving immune function in patients with malignant tumors, particularly those whose immune systems have been weakened due to radiotherapy or chemotherapy. Clinical practice has shown that this formula is remarkably effective in treating esophageal cancer, liver cancer, malignant lymphoma, and aplastic anemia. Specifically, it has completely cured two cases of esophageal cancer, three cases of malignant lymphoma, three cases of aplastic anemia, and two cases of small liver cancer [4]. Xue [5] used the "Lanzhou Formula" to treat primary liver cancer, resulting in a significant reduction in tumor size, complete recovery of liver function, improved quality of life, and prolonged survival. Li et al. [6] conducted a study on the efficacy of combining the Lanzhou Formula with chemotherapy in treating ten cancer patients, finding that the short-term efficacy of the Lanzhou Formula group was superior to that of the chemotherapy-only group (p<0.05), while the incidence of adverse reactions was significantly lower in the Lanzhou Formula group (p<0.01). In addition, when the Lanzhou Formula was used in combination with radionuclide therapy for liver cancer in 15 cases—where the treatment group received both radionuclides and PG, while the control group received only radionuclides—it was found that the treatment group experienced significantly fewer side effects than the control group, and the therapeutic effects of the two groups also differed significantly [7].

In recent years, animal experiments have demonstrated that Peishishengxue Granule can significantly increase the levels of hemoglobin, white blood cells, and platelets in the peripheral blood of aplastic anemia model mice (established through whole-body irradiation with a 3.0 Gy linear accelerator), and can effectively restore the hematopoietic function of the bone marrow in these mice [8]. Histopathological examination of the spleens of these mice revealed that Peishishengxue Granule can alleviate pathological changes in the spleens of aplastic anemia model mice, reduce the rate of lymphocyte apoptosis, promote the restoration of germinal centers, expand the area of the white pulp, thin the capsule, and essentially normalize the dilated sinusoids. Immunohistochemical analysis of CD4 and CD8 levels in the spleens showed that Peishishengxue Granule can increase the CD4 level while decreasing the CD8 level in aplastic anemia model mice, thereby increasing the CD4/CD8 ratio and enhancing the immune function of these mice [9].

Peishishengxue Granule follows the principle of "supporting the body's vital energy and consolidating the root," focusing on overall regulation of the body—specifically, strengthening the immune system and alleviating varying degrees of immune deficiency, thus achieving clinical efficacy. This traditional Chinese medical approach of "supporting the body's vital energy and consolidating the root" shares similarities with modern immunology, molecular biology, and genomics [10–18]. Although previous researchers have made some progress in this field, many aspects still require further exploration.

This experiment establishes a tumor-bearing (H22) mouse model and uses molecular biological techniques to observe the effects of Peishishengxue Granule on the concentration of IL-2 in the supernatant of splenocyte cultures, as well as on the transcription levels of IL-2 mRNA and IFN-γ mRNA in splenocytes. At the same time, the experiment examines the impact of Peishishengxue Granule on the weight of immune organs and the proliferation activity of splenic lymphocytes in tumor-bearing mice, aiming to explore its potential mechanisms of action and provide scientific, microscopic evidence for the widespread clinical application of Peishishengxue Granule.

Experimental Study