Abstract in Chinese

Abstract in Chinese

Objective: To explore the anti-tumor effects and mechanisms of Pei’s Blood-Boosting Granules by observing their impact on H22-bearing mouse tumor cell apoptosis and NF-κB expression.

Method: Using an H22 solid tumor-bearing mouse model, calculate the thymus index (TI) and spleen index (SI) via immune organ weight measurement; calculate the tumor suppression rate of PG; observe the ultrastructure of tumor tissue using transmission electron microscopy; measure the impact of PG on tumor cell NF-κB, cell cycle, and apoptosis rate using flow cytometry.

Results: Animal experiments show: (1) In the high-, medium-, and low-dose groups of Pei’s Blood-Boosting Granules, the thymus index and spleen index of H22-bearing mice were all higher than those of the model group. Specifically, the medium-dose group had TI and SI values of 18.9 mg/10g and 76.1 mg/10g, respectively, representing increases of 36.9% and 19.3% compared with the model group (P<0.05); the high-dose group had TI of 18.1 mg/10g, a 31.2% increase over the model group (P<0.05); (2) The tumor suppression rate of PG was significantly higher than that of the model group, with the medium-dose group showing the best effect at 45.60% (P<0.01); (3) The NF-κB expression in the medium-dose group was 15.74%, significantly lower than that of the model group (P<0.01); (4) The model group had the lowest proportion of cells in G0/G1 phase and the highest proportion in S phase, while the drug-treated groups showed a clear increase in the G0/G1 proportion (P<0.05) and a clear decrease in the S phase proportion (P<0.05); (5) The apoptosis rate in the medium-dose group was 33.46%, significantly higher than that of the model group (P<0.01).

Conclusion: Pei’s Blood-Boosting Granules have anti-tumor effects, with the medium-dose group showing the most pronounced effect. The anti-tumor mechanism of PG is related to enhancing the body’s immune function, influencing gene transcription associated with tumor proliferation and apoptosis, affecting the cell cycle, and promoting tumor cell apoptosis.

Keywords: Pei’s Blood-Boosting Granules; NF-κB; cell cycle; apoptosis rate; tumor suppression rate; flow cytometry Methods: The models of solid tumor were developed by hepatoma cell H22 transplantation in

Research on Pei Zhengxue's Series of Formulas and Medicines

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

Preface

Pei's Hematopoietic Granules is a specialized formula for treating leukemia formulated by Professor Pei Zhengxue, a renowned expert in integrated traditional Chinese and Western medicine in China. It mainly consists of Liuwei Dihuang Decoction combined with Shengmai Powder, plus Taizishen, Beishashen, Dangshen, etc., with heavy use of Taizishen, Dangshen, and Beishashen to tonify the spleen; while Liuwei Dihuang Decoction is used to tonify the kidney. Thirty years ago, this formula completely cured a patient with acute monocytic leukemia (M₅), Ma Changsheng, and was named "Lanzhou Formula" at the Hematology Conference held in Suzhou in 1974 (in 2003, Pei's Hematopoietic Granules successfully cured another leukemia patient (L₂), Liu Ligang, which was reported in both Nanchang Daily and Gansu Daily). In 1997, the formula was made into "Pei's Hematopoietic Granules" as an internal preparation of the Gansu Provincial Academy of Medical Sciences. Since 2002, this preparation has been used as a clinical research project of the Gansu Provincial Academy of Medical Sciences and has passed the achievement appraisal. It also won the First Prize of Huangfu Mi Science and Technology Achievement Award of Gansu Province in 2003.

More than 30 years of clinical application have proven that Pei's Hematopoietic Granules (also known as the "Lanzhou Formula") not only has excellent efficacy against leukemia, but also significantly improves immune function in malignant tumors and in patients with immunosuppression caused by radiotherapy and chemotherapy. Clinical practice has shown that this formula is highly effective against esophageal cancer, liver cancer, malignant lymphoma, aplastic anemia, and other diseases, including complete cure of two cases of esophageal cancer, three cases of malignant lymphoma,

Research on Pei Zhengxue's Series of Formulas and Medicines

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 significant reduction of tumor size, complete recovery of liver function, improved quality of life, and prolonged survival time; Li et al. used the Lanzhou Formula in combination with chemotherapy to treat 10 cancer patients, and the efficacy observation showed that the short-term efficacy of the Lanzhou Formula group was better than that of the chemotherapy-only group (p<0.05); from the perspective of adverse reactions, the Lanzhou Formula group had significantly milder side effects than the chemotherapy-only group (p<0.01). Pei's Hematopoietic Granules was used in combination with radionuclide therapy for 15 cases of liver cancer, with the treatment group receiving radionuclide therapy plus PG, while the control group received only radionuclide therapy, and it was found that the treatment group had significantly milder toxic side effects than the control group, with a significant difference in therapeutic efficacy between the two groups [7].

In recent years, animal experiments have demonstrated that Pei's Hematopoietic Granules can significantly increase the levels of peripheral blood hemoglobin, white blood cells, and platelets in aplastic anemia model mice (established by whole-body irradiation with a 3.0GY linear accelerator), and can effectively restore the hematopoietic function of the bone marrow in these mice [8]. Pathological tissue section observations of mouse spleens show that Pei's Hematopoietic Granules can alleviate pathological changes in the spleens of aplastic anemia model mice, reduce the apoptosis rate of lymphocytes, promote the recovery of germinal centers, expand the white pulp area, thin the capsule, and essentially normalize dilated sinusoids. Immunohistochemical detection of CD4 and CD8 levels in the spleen indicates that Pei's Hematopoietic Granules can increase CD4 levels and decrease CD8 levels in the spleens of aplastic anemia model mice, thereby increasing the CD4/CD8 ratio and enhancing the immune function of these mice [9]. MTT colorimetric assay shows increased lymphocyte proliferation activity; double-antibody sandwich ELISA reveals an upward trend in IL-2 concentration in the supernatant of splenic lymphocyte cultures; reverse transcription-polymerase chain reaction (RT-PCR) demonstrates an upward trend in the expression levels of IL-2 mRNA and IFN-γ mRNA in splenic lymphocytes [10].

Pei's Hematopoietic Granules takes "supporting vital energy and consolidating the root" as its main principle, focusing on overall regulation of the body to varying degrees improve the state of immune deficiency, thus achieving clinical efficacy. This "supporting vital energy and consolidating the root" approach in traditional Chinese medicine shares similarities with modern medical disciplines such as immunology, molecular biology, and genomics [11–17]. Although predecessors have done some work in this area, many aspects still require further exploration.

This experiment establishes a tumor-bearing (H₂₂) mouse model and uses molecular biology techniques to observe the effects of Pei's Hematopoietic Granules on tumor cell apoptosis and NF-κB expression in H₂₂-bearing mice, exploring the anti-tumor effects of Pei's Hematopoietic Granules at the molecular level. The aim is to explore the anti-tumor effects of Pei's Hematopoietic Granules and the mechanisms of herbal medicines in combating tumors at the molecular level. It seeks to provide further experimental evidence for the drug's ability to induce cell apoptosis and regulate cellular signal transduction in anti-tumor treatment, offering sufficient scientific theoretical basis for its widespread clinical application and laying the foundation for developing this preparation into a new type of highly effective traditional Chinese medicine for preventing and treating tumors.

Experimental Study