4.2 Preparation of Tumor-Bearing Mouse Models

4.2 Preparation of Tumor-Bearing Mouse Models

The method used in this experiment to create syngeneic transplant models is to inoculate H22 liver cancer cells into the right anterior axillary region of mice. This method has been widely used in liver cancer research in recent years, offering many advantages, such as stable model creation, direct observation of tumor growth, clear responses in host survival quality and lifespan, and short experimental cycles [112]. After successfully establishing a subcutaneous tumor model in the right anterior axillary region of mice, the formation and proliferation of solid tumors are observed. Subcutaneous transplant tumor models are easy to observe and intervene in, convenient for inoculation, and have a high success rate.

Research on Pei Zhengxue’s Series of Formulas and Medications

4.3 Selection of Positive Control Drugs

Positive control drugs are often chosen for clinical trials of new traditional Chinese medicines. A positive drug control trial compares the test drug with a known active and effective drug [114]. There are two basic principles for selecting positive control drugs: comparability within the same category and recognized effectiveness [115], meaning searching for and testing drugs in the Chinese Pharmacopoeia and ministry-issued standards that have similar indications, functions, dosage forms, and routes of administration [116]; and choosing drugs currently recognized by academia or society as safe and effective [117].

Compound Cantharid Capsules are a traditional Chinese medicine preparation with cantharid as the main ingredient, possessing functions such as breaking blood stasis, eliminating blood clots, fighting cancer, and detoxifying. Zhang Jianwu [118] and others have studied and proposed that the active ingredient cantharid in Compound Cantharid Injection has a good in vivo anti-tumor effect on mouse H22 tumors. Studies have shown that Compound Cantharid Capsules can improve the proportion of T-cell subsets, enhance the body’s cellular immunity, and induce tumor cell apoptosis; their mechanism of action may involve increasing the activity of NK and LAK cells in tumor-bearing mice and raising IL-2 and TNF-α levels [119].

In this experiment, Compound Cantharid Capsules were selected as the positive control drug because their functions and modern pharmacological research are quite similar to Pei’s Soft Liver and Anti-Bloating Pill.

4.4 Anti-Tumor Effects of Pei’s Soft Liver and Anti-Bloating Pill on Tumor-Bearing Mice

The results of this experiment show that the average tumor weight in the low-, medium-, and high-dose groups of Pei’s Soft Liver and Anti-Bloating Pill was lower than that of the model control group, at (0.935±0.227) g, (0.776±0.122) g, and (0.926±0.237) g respectively, all statistically significant compared with the model control group (p<0.05), with anti-tumor rates of 23.9%, 36.8%, and 24.5% respectively. This indicates that Pei’s Soft Liver and Anti-Bloating Pill has a good effect in inhibiting tumor growth.

4.5 Effects of Pei’s Soft Liver and Anti-Bloating Pill on Immune Organs of Tumor-Bearing Mice

As is well known, the thymus and spleen are the most important immune organs in the human body. The thymus is the main site for the development, differentiation, and maturation of T cells, which play an extremely important role in the body’s cellular immunity and immune regulation. The spleen is the key location where lymphocytes migrate and mount immune responses upon antigen stimulation, producing immune effector molecules and performing vital immune functions. In a sense, the weight of the thymus and spleen can reflect the body’s immune function, so their weights can be used as preliminary indicators for observing immune function [120].

The experimental results show that the thymus and spleen of the model control group mice shrank in size and weight, with some thymuses having unclear lobes and appearing grayish-white, and the spleens turning pale red. The thymus index of mice in each PR-GXP dose group was higher than that of the model control group, with the medium-dose group’s TI reaching (53.2±15.6) mg/10 g, statistically significant compared with the model control group (p<0.05); compared with the blank group, the model control group’s thymus index decreased (p<0.05). The spleen index of mice in each PR-GXP dose group was also higher than that of the model control group, statistically significant compared with the model control group (p<0.05); compared with the blank group, the model control group’s spleen index decreased (p<0.05). The experimental results indicate that Pei’s Soft Liver and Anti-Bloating Pill significantly enhances the thymus and spleen indices of tumor-bearing mice, thereby improving their non-specific immune function.

4.6 Pei’s Soft Liver and Anti-Bloating Pill Inhibits Cell Proliferation and Promotes Cell Apoptosis