Chinese Abstract

Chinese Abstract

Objective: By observing the effect of Pei’s Blood-Boosting Granules (PG) on the expression of VEGF in tumor tissue and serum IL-12 levels in H₂₂ tumor-bearing mice, this study aims to explore the pharmacological mechanism of PG in treating tumors and provide a scientific basis for its antitumor effects.

Methods: A solid tumor model was established by transplanting H₂₂ hepatoma cells into mice, which were then divided into a blank group, a model group, a high-dose PG group, a medium-dose PG group, a low-dose PG group, and a ZG group. After 10 days of medication, the tumor inhibition rate, thymus index (TI), and spleen index (SI) were calculated. Immunohistochemistry was used to determine the expression of VEGF in the tumor tissue of the bearing mice. ELISA was employed to measure the concentration of IL-12 in the serum of the bearing mice.

Results:

1. The high-, medium-, and low-dose PG groups all showed significant inhibition of tumor weight in the H₂₂ tumor-bearing mice, with the average tumor weight significantly different from that of the model group (p<0.01). There was also a significant difference in average tumor weight between the high- and medium-dose groups and the low-dose group (p<0.05 or p<0.01). The tumor inhibition rates for the high-, medium-, and low-dose groups were 36.26%, 37.56%, and 24.91%, respectively.
2. The thymus index and spleen index of the mice in the various PG dose groups were higher than those of the model group. The thymus index of the high-, medium-, and low-dose groups differed significantly from that of the model group (p<0.05 or p<0.01). Similarly, the spleen index of the mice in the various PG dose groups also differed significantly from that of the model group (p<0.05 or p<0.01).
3. The number of VEGF-positive cells in the tumor tissue of the mice in the various PG dose groups was lower than that of the model group to varying degrees, with the high- and medium-dose groups showing significant differences compared with the model group (p<0.05).
4. The serum IL-12 concentrations in the mice of the various PG dose groups were higher than those of the model group, with the high- and medium-dose groups showing significant differences compared with the model group (p<0.05).

Conclusion: This experiment demonstrates that PG has a significant inhibitory effect on the growth of H₂₂ tumors in mice. PG increases the weight of the immune organs—thymus and spleen—in tumor-bearing mice, thereby enhancing the body's nonspecific immune function. Furthermore, PG markedly suppresses the expression of VEGF in tumor tissue while simultaneously increasing the serum IL-12 levels. These findings suggest that one of the mechanisms underlying PG's antitumor effects may be downregulating VEGF expression in tumor tissue and upregulating serum IL-12 levels, thus inhibiting tumor angiogenesis and achieving an antitumor effect.

Keywords: Pei’s Blood-Boosting Granules (PG); H₂₂ (liver cancer); vascular endothelial growth factor (VEGF); interleukin-12 (IL-12)

Research on Pei Zhengxue’s Series of Formulas and Medicines

ABSTRACT

Objective: By observing the effect of Peishishengxue Granules (PG) on the expression of VEGF in tumor and IL-12 in serum on H22 bearing mice to explore the mechanism of PG treatment of cancer medicine and to provide a scientific basis for the antitumor effects of the drug.

Methods: The models of solid tumor were developed by hepatoma cell H22 transplantation in mice. Divided into: normal group, model group, PG high-dose group, PG medium-dose group, PG low-dose group and ZG group. Except the normal and model groups were dealt with drug-intervention. After ten days, all mice were killed. The weights of solid tumor, thymus and spleen were measured. The tumor inhibiting rates, the thymus index (TI) and the spleen index (SI) were calculated. Assessing effects of PG on VEGF in tumor by immunohistochemistry method. Detected the concentration of IL-12 in serum by ELISA.

Results:

1. PG high, medium and low-dose group on H22 tumor-bearing mice showed significant inhibition of tumor weight. The average tumor weight compared with the model group were significantly different (p<0.01). The average tumor weight of PG high and medium-dose group compared with PG low-dose group have significant difference (p<0.05, or p<0.01). PG high, medium and low-dose group tumor inhibition rates were 36.26%, 37.56%, 24.91%.

2. The thymus index and spleen index of tumor-bearing mice dealt with PG various doses were higher than model group. The thymus index of PG high, medium and low-dose mice compared with model group significant difference (p<0.05, or p<0.01). PG spleen index of mice with various doses compared with model group significant difference (p<0.05, or p<0.01).

3. The expression of VEGF in tumor on PG different dose groups have less than the model group. PG high and medium-dose group compared with the model group were significantly different (p<0.05).

4. PG with various doses of IL-12 in serum concentrations higher than those in model group. PG high and medium-dose group compared with the model group were significantly different (p<0.05).

Conclusion: PG had remarkable restrain effect on tumor growth in H22 tumor-bearing model mice. PG has the function of weightening the immune organs (the thymus, the spleen). PG can improve the body's non-specific immune function. Experimental results suggest that: PG antitumor effect may be one of the mechanisms by inhibiting the expression of VEGF in tumor and regulating the content of IL-12 in serum.

Key words: Peishishengxue Granule (PG); transplanted liver cancer (H22); vascular endothelial cell growth factor (VEGF); interleukin-12 (IL-12)

Research on Pei Zhengxue’s Series of Formulas and Medicines

Foreword

Today, malignant tumors have become one of the most serious diseases threatening human life and health. Currently, the main treatments for tumors are surgery, radiotherapy, and chemotherapy. Surgical treatment can only remove tumors visible to the naked eye, but it is powerless against recurrence and metastasis. Meanwhile, radiotherapy and chemotherapy, while killing tumor cells, also damage normal cells, especially suppressing bone marrow hematopoiesis and weakening the body's immune function. With the development of traditional Chinese medicine, continuous research has revealed that TCM offers multiple pathways, multi-target effects, fewer toxic side effects, and a lower likelihood of drug resistance in tumor treatment. In particular, TCM excels at addressing the multifactorial and multistep nature of tumor pathogenesis. The antitumor effects of Chinese herbal medicines are increasingly recognized and have become an important adjunct in tumor therapy.

Professor Pei Zhengxue, drawing on his more than forty years of clinical experience, believes that "deficiency of vital energy" is the root cause of the occurrence and development of malignant tumors; strengthening vital energy and consolidating the body's foundation is the fundamental principle in treating malignant tumors, with "tonifying the spleen and kidney" as its essence. Based on this principle, he formulated the "Lanzhou Formula," which, after more than forty years of continuous practice, refinement, and reorganization, eventually evolved into "Pei’s Blood-Boosting Granules." Over the years, this medicine has achieved satisfactory clinical results in tumor treatment, often used in conjunction with preoperative, postoperative, and radiochemotherapy regimens to enhance efficacy, reduce toxicity, and boost immunity. Moreover, for patients with advanced-stage tumors who have missed the opportunity for surgery, radiotherapy, or chemotherapy, it can alleviate symptoms and improve quality of life. The purpose of this experimental study is to objectively demonstrate the antitumor and anti-metastatic effects of Pei’s Blood-Boosting Granules, explore their mechanisms of action, and provide a scientific basis for the widespread clinical application of this medicine.

I. Rationale for the Study

Pei's Hematopoietic Granules is a specialized formula for treating leukemia, formulated by Professor Pei Zhengxue, a renowned expert in integrated Chinese and Western medicine and doctoral supervisor, based on more than forty years of clinical experience [1]. The overall principle of the formula is to strengthen the spleen and tonify the kidney, supporting the body's vital energy and consolidating its foundation [2]. Due to its complete and thorough cure of a patient with acute monocytic leukemia (M₅), Ma Changsheng, thirty years ago, the formula was named "Lanzhou Formula" at the 1974 National Hematology Conference in Suzhou [3]. In 2003, Pei's Hematopoietic Granules successfully cured another leukemia patient (L₂), Liu Ligang, which was subsequently reported in both the Nanchang Daily and the Gansu Daily. In 1997, the formula was designated as an internal preparation of the Gansu Provincial Academy of Medical Sciences and named "Pei's Hematopoietic Granules." Some experimental studies on this formulation have passed the evaluation of scientific achievements and have won the First Prize of the Huangfu Science and Technology Achievement Award of Gansu Province.

Animal experiments have demonstrated that Pei's Hematopoietic Granules significantly increase the weight of immune organs—thymus and spleen—in tumor-bearing mice, thereby enhancing their nonspecific immune function. The granules can promote the proliferation of splenic lymphocytes stimulated by ConA, strengthening the cellular immune function of tumor-bearing mice and improving their anti-tumor effects. They also stimulate the secretion of IL-2 and the expression of IL-2 and IFN-γ mRNA, suggesting that Pei's Hematopoietic Granules regulate the body's immune function by influencing cytokines [4]. Furthermore, Pei's Hematopoietic Granules can modulate the body's dysregulated immune status, enhancing delayed-type hypersensitivity reactions in tumor-bearing mice, increasing their cellular immune function, boosting the phagocytic activity of peritoneal macrophages, and causing an upward trend in cytokines such as IL-1, IFN-γ, and NO [5]. Additionally, Pei's Hematopoietic Granules exhibit a significant inhibitory effect on tumors in tumor-bearing mice. The granules can markedly elevate peripheral blood hemoglobin, white blood cell count, and platelet levels in aplastic anemia model mice (established through whole-body irradiation with a 3.0 Gy linear accelerator), effectively restoring the hematopoietic function of bone marrow in these models [6]. Immunohistochemical analysis of CD4⁺ and CD8⁺ levels in the spleen shows that Pei's Hematopoietic Granules tend to increase CD4⁺ levels while decreasing CD8⁺ levels in aplastic anemia model mice, thus raising the CD4⁺/CD8⁺ ratio and enhancing their immune function [7]. Moreover, Pei's Hematopoietic Granules can significantly inhibit mutant P53 and upregulate Caspase-3 protein expression, indicating that the drug may induce apoptosis of tumor cells by downregulating mutant P53 and upregulating Caspase-3 protein expression [8]. Pei's Hematopoietic Granules also suppress NF-κB expression, affecting gene transcription and blocking the progression of tumor cells from G1 phase to S phase, thereby inducing apoptosis [9]. In addition, Pei's Hematopoietic Granules exhibit a certain antagonistic effect against the suppression of immune organs—thymus and spleen—caused by 5-FU, maintaining the body's immune function by increasing the weight of these organs and resisting damage from external stimuli [10]. After chemotherapy, Pei's Hematopoietic Granules result in higher concentrations of TNF-α and IFN-γ compared with the 5-FU-only group, regulating the body's immune capacity and thereby inhibiting tumor growth [11]. Xue et al. [12] used the "Lanzhou Formula" to treat primary liver cancer, achieving significant reduction in tumor size, complete recovery of liver function, improved quality of life, and prolonged survival; Li et al. [13] conducted an efficacy observation on ten cases of cancer treated with the Lanzhou Formula combined with chemotherapy, showing that the short-term efficacy of the Lanzhou Formula group was superior to that of the chemotherapy-only group (p<0.05); moreover, the incidence of adverse reactions was significantly lower in the Lanzhou Formula group than in the chemotherapy-only group (p<0.01). When Pei's Hematopoietic Granules were used in combination with radionuclide therapy for liver cancer in fifteen cases, the treatment group received radionuclide therapy plus Pei's Hematopoietic Granules (PG), while the control group received only radionuclide therapy, revealing that the treatment group experienced significantly fewer side effects than the control group, and there was also a significant difference in therapeutic efficacy between the two groups [14]. During my apprenticeship under the guidance of my mentor, according to incomplete statistics, this formula has cured three cases of esophageal cancer, one case of gastric cancer, three cases of leukemia, three cases of aplastic anemia, and three cases of primary liver cancer [15]. Notably, it has shown remarkable efficacy in treating esophageal cancer, gastric cancer, liver cancer, leukemia, and aplastic anemia.

Given its significant clinical efficacy, Pei's Hematopoietic Granules has become a well-known specialized remedy for hematological and oncological diseases both within and outside the province. Although considerable experimental research data are already available on this formulation, further studies at the molecular biology level remain insufficient. Recent medical research indicates that tumor angiogenesis is the pathological basis for tumor growth and metastasis. Vascular endothelial growth factor (VEGF) is a large-molecule bioactive substance discovered in recent years that plays an important biological role in changes in vascular permeability, proliferation of vascular endothelial cells, and angiogenesis, closely related to tumor growth, invasion, and metastasis. Interleukin-12 (IL-12) is an immunomodulatory factor and also an angiogenesis inhibitor, possessing multiple biological functions. IL-12 plays an important role in the prevention and treatment of tumors, infections, autoimmune diseases, and other conditions, capable of inhibiting tumor angiogenesis, preventing tumor growth and metastasis, and improving the body's immune function. This experiment aims to establish a mouse H₂₂ model to observe the effects of Pei's Hematopoietic Granules on VEGF expression in H₂₂ tumor tissue and serum IL-12 levels, thereby exploring the anti-tumor effects of Pei's Hematopoietic Granules and the anti-tumor mechanisms of traditional Chinese medicine from the perspective of tumor angiogenesis.

II. Experimental Study