Chinese Abstract

Chinese Abstract

Objective: To observe the effects of Pei’s Shengxue Granules (PG) on tumor cell apoptosis and the expression of mutant P53 and Caspase-3 proteins, and to analyze their mechanisms of action, thereby providing a scientific basis for the drug’s tumor-suppressing effects.

Methods: Using a solid tumor model in H22 (liver cancer) bearing mice, we observed PG’s inhibitory effect on H22 tumors, measured PG’s impact on P53 expression in H22 tumor cells via immunohistochemistry, and assessed PG’s influence on Caspase-3 protein expression in H22 tumor cells using flow cytometry.

Results: All dosage groups of PG—low, medium, and high—showed greater inhibition of H22 tumors in tumor-bearing mice than the model group. The low-dose group had an inhibition rate of 29.3%, which was significantly different from the model group (P<0.05). The medium- and high-dose groups had inhibition rates of 35.9% and 36.8%, respectively, both significantly higher than the model group (P<0.01). Pathological tissue section observations of mouse tumors showed that PG could promote necrosis of H22 tumor tissue in model mice. The thymus index and spleen index of tumor-bearing mice in all PG dosage groups were higher than those of the model group, with the high-dose group showing TI and SI values of 36.0 mg/10 g and 64.4 mg/10 g, respectively—increases of 29.0% and 20.4% compared with the model group (P<0.01); the medium-dose group had a TI of 35.2 mg/10 g, a 26.2% increase over the model group (P<0.05). The expression of mutant P53 in tumor cells of all PG dosage groups was lower than that of the model group, with the medium- and high-dose groups showing the best results; the average number of positive cells for mutant P53 was 54.25 and 55.38, respectively, both significantly different from the model group (P<0.05). The expression of Caspase-3 protein in tumor cells of all PG dosage groups was higher than that of the model group, with the high- and medium-dose groups showing significant differences compared with the model group (P<0.01).

Conclusion: This experiment demonstrates that PG has a significant inhibitory effect on the growth of H22 tumors in mice, and pathological tissue section observations reveal that PG can promote necrosis of H22 tumor tissue in model mice. PG also has a weight-increasing effect on the immune organs—thymus and spleen—of H22 tumor-bearing mice. Furthermore, PG induces apoptosis in liver cancer cells and, after acting on tumor cells, significantly downregulates mutant P53 (Mt) while promoting the expression of Caspase-3. This suggests that one of the mechanisms underlying PG’s anti-tumor effects may be through suppressing mutant P53 and upregulating Caspase-3 protein expression. PG may therefore have broad application prospects in tumor treatment.

Keywords: Pei’s Shengxue Granules, H22 (liver cancer), cell apoptosis, mutant p53, Caspase-3

Research on Pei Zhengxue’s series of prescriptions

ABSTRACT

Objective: To provide scientific evidence of PG on tumor cell inhibiting through observing the effects of PG on tumor cell apoptosis, the effects on Mild type P53, caspase-3 protein expression and analyzing its effect and theories.

Methods: The models of solid tumor were developed by hepatoma cell H22 transplantation in mice. Experiment parameters were obtained by observing PG's inhibiting on hepatoma cell H22, assessing effects of PG on cell H22 P53's expression by immune histochemistry method; checking the effects of PG on cell H22's caspase-3 protein expression by Flow cytometry method.

Results: PG had remarkable restrain effect on tumor growth in H22 tumor-bearing model mice. The inhibitory effect of each group was higher than the model group. The restrain effect of low group was 29.3%, it has remarkable difference than the control group (p<0.05). The restrain effect of medium and high group was 35.9% and 35.9%, it has remarkable difference than the control group (p<0.01). The TI and SI was increased in PG groups. In high dose group they were significantly increased (p<0.01), and rose by 29.0% and 20.4% respectively; The TI in medium dose group was 18.0mg/10g, and rose by 26.2% (p<0.05). The express of Mild type p53 in every group of PG was lower than control group, and the medium group and the high group was very obvious, they number positive cell was 54.25 and 55.38, it has remarkable difference than the control group (p<0.05). The caspase-3 protein expression in each group of PG was increased than control group, the medium group and the high group has remarkable difference than the control group (p<0.05).

Conclusion: PG had remarkable restrain effect on tumor growth in H22 tumor-bearing model mice. PG make H22 tumor organize rotten. PG has the function of weightening the immune organs (the thymus, the spleen), PG has the effect of make H22 cell death, PG make Mild type p53 descend and impel the expression of caspase-3 protein.

Keywords: Peishishengxue Granule (PG), H22, apoptosis, Mild type p53, caspase-3

Foreword

Peishishengxue Granule is a specialized prescription for treating leukemia formulated by Professor Pei Zhengxue, a nationally renowned expert in integrated traditional Chinese and Western medicine, based on more than forty years of clinical experience and the theory of reinforcing vital energy and consolidating the root. Building upon the holistic view of traditional Chinese medicine, Professor Pei employs the principle of reinforcing vital energy and consolidating the root to develop various herbal prescriptions combined with chemotherapy for leukemia treatment. His approach respects tradition while innovating, resulting in the "Lanzhou Formula," which has demonstrated remarkable clinical efficacy. The "Lanzhou Formula" gained its name at the 1974 National Hematology Conference in Suzhou after completely curing a patient with acute monocytic leukemia (M5) named Ma Changsheng thirty years earlier. In 2003, Peishishengxue Granule successfully treated another leukemia patient (L2) named Liu Ligang, following previous successes in the 1980s...

Research on Pei Zhengxue’s series of formulas and medicines

This research has been reported by the Nanchang Daily and the Gansu Daily. In addition to treating leukemia, Professor Pei has also used this formula to treat various cancers and refractory anemias, achieving significant therapeutic effects in esophageal cancer, gastric cancer, myelodysplastic syndromes, aplastic anemia, and other conditions.[3][4] In 1997, the formula was designated as an internal preparation of the Gansu Provincial Academy of Medical Sciences and renamed "Peishishengxue Granule." Some experimental studies on this formulation have already passed evaluation and received awards, including the First Prize for Scientific and Technological Achievements from Huangfu Mi in Gansu Province. Due to its outstanding clinical efficacy, Peishishengxue Granule has so far treated tens of thousands of patients both within and outside the province, earning recognition from clinicians and patients alike and gaining considerable fame domestically and internationally.

According to rough recollections, this formula 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] Notably, it has shown remarkable efficacy in treating esophageal cancer, liver cancer, malignant lymphoma, and aplastic anemia. For instance, Xue[5] used the "Lanzhou Formula" to treat primary liver cancer, resulting in a significant reduction in tumor size, complete restoration of liver function, improved quality of life, and prolonged survival. Li et al.[6] conducted a study on ten cancer patients treated with the Lanzhou Formula in combination with chemotherapy, 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). Furthermore, when Peishishengxue Granule was used in conjunction with radionuclide therapy for fifteen liver cancer patients—where the treatment group received both radionuclides and Peishishengxue Granule, while the control group received only radionuclides—the treatment group exhibited markedly milder side effects, and the therapeutic outcomes were also significantly better compared to the control group.[7]

Animal experiments have demonstrated that Peishishengxue Granule significantly increases the weight of immune organs such as the thymus and spleen in tumor-bearing mice, thereby enhancing their non-specific immune functions. It can promote the proliferation of splenic lymphocytes stimulated by ConA, strengthen cellular immunity in tumor-bearing mice, and enhance anti-tumor effects. Additionally, it stimulates the secretion of IL-2 and the expression of IL-2 and IFN-γ mRNA, suggesting that Peishishengxue Granule regulates immune function by influencing cytokines.[8] Peishishengxue Granule can also regulate disordered immune states, increasing delayed-type hypersensitivity responses in tumor-bearing mice and boosting their cellular immunity. It enhances the phagocytic function of peritoneal macrophages and leads to increased levels of cytokines such as IL-1, IFN-γ, and NO. Moreover, Peishishengxue Granule exhibits a clear inhibitory effect on tumors in H22-bearing mice. Strengthening immune function is a key mechanism underlying its anti-tumor activity.[9] Peishishengxue Granule can significantly elevate peripheral blood hemoglobin, white blood cell count, and platelet levels in mice with aplastic anemia models (established through whole-body irradiation with a 3.0 Gy linear accelerator), effectively restoring bone marrow hematopoietic function in these models.[10] Histopathological examination of mouse spleen tissue reveals that Peishishengxue Granule alleviates pathological changes in the spleen of aplastic anemia models, reduces lymphocyte apoptosis, promotes the recovery of germinal centers, expands the white pulp area, thins the capsule, and restores dilated sinusoids to normal. Immunohistochemical analysis of CD4 and CD8 levels in the spleen shows that Peishishengxue Granule increases CD4 levels while decreasing CD8 levels in aplastic anemia models, raising the CD4/CD8 ratio and thus enhancing immune function.[11] These findings are consistent with the long-term clinical efficacy of Peishishengxue Granule in extending survival and slowing tumor growth. Despite the efforts of colleagues in researching and developing Peishishengxue Granule, microscopic investigations into this clinically proven, highly effective formula remain insufficient. Therefore, building on previous studies, we employed the H22 solid tumor mouse model to explore Peishishengxue Granule's anti-tumor effects and related mechanisms. By observing how different doses of PG affect the growth of H22 tumors in mice, we determined its anti-tumor efficacy. We also investigated the anti-tumor mechanisms of PG from the perspective of inducing tumor cell apoptosis, examining protein and gene-level expressions of P53 and Caspase-3 in H22 tumor cells, and exploring the molecular mechanisms behind Peishishengxue Granule's ability to induce tumor cell apoptosis. This research aims to provide scientific evidence for the use of this formula in treating malignant tumors and for integrating traditional Chinese medicine with Western medicine in oncology, thereby offering a scientific and detailed basis for the widespread clinical application of Peishishengxue Granule.

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Experimental Study