4.3 Effects of Pei’s Soft Liver and Anti-Bloating Pill on Cytokines

4.3 Effects of Pei’s Soft Liver and Anti-Bloating Pill on Cytokines

Cytokines are small protein molecules with important biological activity secreted by activated immune cells and related cells (such as fibroblasts and endothelial cells), playing a crucial role in the initiation, propagation, and regulation of immune responses, even serving as a central regulatory factor. In addition to their individual multiple biological activities, cytokines also interact with each other. Modern research shows that IFN-γ is mainly secreted by activated T cells and natural killer cells, and participates in immune responses in various ways. TNF-α is produced by activated macrophages/monocytes, NK cells, and fibroblasts, and is currently considered the cytokine with the strongest anti-tumor effect [19], possessing systemic effects, primarily participating in inflammatory responses, regulating immune responses, and exerting anti-tumor effects. IFN-γ and TNF-α, as specific indicators reflecting the body’s immune function, participate in the entire process of tumor occurrence, development, and demise.

4.3.1 Effects on Tumor Necrosis Factor (TNF-α)

TNF-α is a cytokine produced by activated macrophages/monocytes, NK cells, and fibroblasts, possessing multiple biological functions such as mediating inflammation, regulating immunity, and exerting anti-tumor effects. TNF-α has a direct cytotoxic and apoptosis-inducing effect on tumor cells [20–21]. An important characteristic of TNF-α’s anti-tumor effect is that it only exerts cytotoxicity on tumor cells without harming normal cells, demonstrating highly selective killing power [22]. Its mechanism of action involves binding to TNF receptors on the cell surface, generating reactive oxygen species and peroxides, leading to cell lysis. At the same time, TNF-α can also promote the synthesis and secretion of various anti-tumor cytokines (such as GM-CSF, IL-1, IL-2, and IL-6, etc.), thereby enhancing the anti-tumor immune defense of cancer patients and indirectly exerting multiple anti-tumor immune effects.

In this experiment, ELISA was used to measure the concentration of TNF-α in the serum of tumor-bearing mice, in order to assess the therapeutic effect of Pei’s Soft Liver and Anti-Bloating Pill on tumors and its immune-regulating effect on the body. The results showed that the serum TNF-α concentration in the Blister Beetle group increased, which was statistically significant compared with the Model Control Group (P<0.05); the serum TNF-α concentrations in all PRGXP dosage groups were higher than those of the Model Control Group, with the medium-dose group reaching (85.97±19.41) pg·ml⁻¹, significantly higher than the Model Control Group (P<0.05), and moreover, the medium-dose group’s TNF-α concentration was also higher than that of the positive drug control group (Blister Beetle group) (P<0.05). This suggests that Pei’s Soft Liver and Anti-Bloating Pill can enhance the body’s immunity and increase the concentration of TNF-α in the serum, thereby achieving the ultimate goal of anti-tumor treatment.

4.3.2 Effects on Interferon (IFN-γ)

IFN-γ is mainly secreted by activated T cells and natural killer cells, and participates in immune responses in various ways, being one of the most potent inflammatory mediators in the body. Its biological effects are very broad, capable of triggering inflammatory responses through multiple mechanisms and playing an important role in tumor treatment. The main mechanisms by which IFN-γ inhibits tumor cell proliferation are as follows: ① It can inhibit angiogenesis within the tumor [10]; ② It has a direct cytotoxic effect on malignant tumor cells; ③ Interferon can affect the mitotic cycle of tumor cells, inducing apoptosis [11]; ④ It can exert anti-tumor activity by affecting the extracellular matrix [13].

In this experiment, ELISA was used to measure the concentration of IFN-γ in the serum of tumor-bearing mice, in order to assess the therapeutic effect of Pei’s Soft Liver and Anti-Bloating Pill on tumors and its immune-regulating effect on the body. The results showed that the serum IFN-γ concentration in the Blister Beetle group increased, which was statistically significant compared with the Model Control Group (P<0.05); the serum IFN-γ concentrations in the medium- and high-dose groups of Pei’s Soft Liver and Anti-Bloating Pill increased to varying degrees, but all were higher than those of the Model Control Group, with the medium-dose group reaching (90.85±11.91) pg·ml⁻¹, higher than the Model Control Group (P<0.05). Moreover, the medium-dose group’s IFN-γ concentration was also higher than that of the positive drug control group (Blister Beetle group) (P<0.05), suggesting that Pei’s Soft Liver and Anti-Bloating Pill can enhance the body’s immunity and increase the concentration of IFN-γ in the serum, thereby achieving the ultimate goal of anti-tumor treatment.

4.4 Selection of Positive Control Drugs

According to the principles for selecting positive drugs [25]: ① Comparability: its functions, indications, dosage forms, and routes of administration should be similar to those of the new drug; ② Legality: the positive drug must be a legally approved medication listed in the Pharmacopoeia, a ministerial standard, or newly approved for production; ③ Preference for the best: the selected drug should be a representative “good drug” that is currently recognized as effective by the academic community or society. Cantharidin—pungent in taste, warm in nature, and enters the Liver, Kidney, and Stomach meridians. It has the effects of breaking blood stasis, eliminating tumors, attacking toxins, and dispersing nodules. Its active ingredient is cantharidin, which can inhibit the growth of tumor cells and induce apoptosis [26]. Experiments have shown that it first inhibits protein synthesis in cancer cells, then affects the biosynthesis of RNA and DNA, ultimately blocking the growth and division of cancer cells. The main component of Compound Cantharidin Capsules is cantharidin. Studies have demonstrated that these capsules block the M phase of the cancer cell division cycle, disrupt the cytoskeleton and ultrastructure of cancer cells, and increase the rate of respiratory inhibition and lysosomal enzyme activity in cancer cells [27]. Zhang Jianwu [28] et al. suggested that Compound Cantharidin Injection exhibits good in vivo anti-tumor effects against H₂₂ tumors in mice, attributing this effect to the direct action of cantharidin, the active ingredient in the injection.

The Compound Cantharidin Capsules used in this experiment share considerable similarities with Pei’s Soft Liver and Anti-Bloating Pills in terms of efficacy and modern pharmacological research; therefore, they were selected as the positive control drug.

4.5 Modern Research on Single Herbs in Pei’s Soft Liver and Anti-Bloating Pills

To explore the mechanisms by which traditional Chinese medicine treats primary liver cancer, scholars both domestically and internationally have conducted extensive experimental studies. Professor Pei Zhengxue’s prescription for treating liver cancer—Pei’s Soft Liver and Anti-Bloating Pills—has been studied experimentally, revealing that its potential anti-cancer mechanisms may involve directly killing and inhibiting liver cancer cells, as well as protecting and regulating the body’s immune function. Specifically:

• Scutellaria barbata: pungent and bitter, cold in nature. Enters the Lung, Liver, and Kidney meridians. Functions and indications include clearing heat and detoxifying, resolving blood stasis, and promoting diuresis. Zhang Nina et al. [29] found that Scutellaria barbata can effectively inhibit tumor angiogenesis, possibly through mechanisms such as blocking endothelial cell migration and downregulating VEGF protein expression.

• Astragalus membranaceus: has the effects of tonifying qi, strengthening defensive qi, and supporting sores. Modern pharmacological research shows that Astragalus can enhance stress resistance in mice, boost immune function, and accelerate the repair of bodies damaged by radiation. Its contained astragalosides, polysaccharides, trace elements, and other beneficial components can increase lymphocyte proliferation in tumor-bearing mice and enhance NK cell activity, thereby regulating immune function and achieving an anti-tumor effect [30]. According to “Zhenzhuang,” Astragalus can tonify all deficiencies and replenish vital energy.

• Hedyotis diffusa: has the effect of clearing heat and detoxifying; modern research proves that it can kill liver cancer cells and significantly enhance the body’s immunity.

• Laminaria japonica: salty and cold, enters the Liver and Kidney meridians, with the effects of eliminating phlegm and softening hard masses. As recorded in “Compendium of Materia Medica”: “Laminaria is salty, which moistens downward, and cold, which clears heat and draws out water; thus it can eliminate goiters, tubercles, and stubborn accumulations such as yin ulcers.” Its laminarin polysaccharides can indirectly inhibit or kill tumor cells by enhancing the body’s immunity [32].

• Laminaria japonica: salty and cold, enters the Liver and Kidney meridians, with the effects of eliminating phlegm and softening hard masses. Modern research shows that laminarin polysaccharides and nucleic acids in Laminaria japonica can activate macrophages, thereby inhibiting tumor cell proliferation and killing tumors [33].

• Bupleurum chinense: enters the Liver and Gallbladder meridians, and its extracts have a significant inhibitory effect on tumors [34–35]. Modern research indicates that Bupleurum has good effects in protecting the liver from damage, promoting bile flow, and lowering transaminase levels.

• Paeonia lactiflora: bitter, sour, sweet, and slightly cold in nature, entering the Liver and Spleen meridians. It nourishes blood, regulates menstruation, calms the Liver and relieves pain, and astringes yin to stop sweating.

In summary, Pei’s Soft Liver and Anti-Bloating Pills are a proven prescription for treating primary liver cancer, with some of the herbs in the formula indeed having anti-cancer effects. However, Pei’s Soft Liver and Anti-Bloating Pills are an organic whole for treating liver cancer; their therapeutic effect does not stem solely from a few anti-cancer herbs, but rather from overall regulation and activation of the body’s own immune function. As Professor Pei himself said: “Primary liver cancer is fundamentally deficiency with superficial excess—it appears locally, but is actually a manifestation of a systemic disease, a condition of overall deficiency with localized excess. Therefore, in treatment, I propose strengthening the root to dissipate the accumulation, and dissipating the accumulation to consolidate the root.” Anti-cancer drugs only treat the symptoms, while strengthening the root and consolidating the foundation is the core principle of this prescription.

Research on Pei Zhengxue’s series of prescriptions

Conclusion