Chinese Abstract

Chinese Abstract

Objective: To observe the effect of Peishiruanganxiaopiwan (PRGXP) on serum TNF-α and IFN-γ in H₂₂ tumor-bearing mice, and to explore the regulatory effect of PRGXP on the immune system of tumor-bearing mice as well as its inhibitory effect on tumors.

Methods: A tumor-bearing mouse model was established by subcutaneous inoculation at the right anterior axilla of mice. The mice were randomly divided into a blank group, a model control group, a high-dose PRGXP group, a medium-dose PRGXP group, a low-dose PRGXP group, and a Fufang Banmao capsule (BM) group. The drugs were administered by gavage for 10 days. Twenty-four hours after the last dose, the mice were weighed, their eyeballs were removed for blood collection using clean test tubes, the blood was allowed to clot at room temperature for 2 hours, then centrifuged at 1500 rpm for 10 minutes. The serum was collected, packaged, and stored at −20°C. The mice were euthanized by spinal dislocation, and the tumor tissue, thymus, and spleen were dissected out and weighed. The tumor inhibition rate, thymus index (TI), and spleen index (SI) were calculated. The concentrations of TNF-α and IFN-γ in the serum of tumor-bearing mice were measured using the ELISA method.

Results:

① The average tumor weights in the high-, medium-, and low-dose PRGXP groups were all lower than those in the model control group, at (0.926±0.237)g, (0.776±0.122)g, and (0.935±0.227)g, respectively, all showing statistically significant differences compared with the model control group (P<0.05). The corresponding tumor inhibition rates were 24.5%, 36.8%, and 23.9%.

② The thymus index (TI) and spleen index (SI) in the high-, medium-, and low-dose PRGXP groups were all higher than those in the model control group. Specifically, the TI of the medium-dose group was (53.2±15.6)mg/10g, an increase of 31.4% compared with the model control group, which was statistically significant (P<0.05); the SI of the medium-dose group was (65.0±10.1)mg/10g, an increase of 45.1% compared with the model control group, also statistically significant (P<0.05).

③ The concentrations of TNF-α in the high-, medium-, and low-dose PRGXP groups were all higher than those in the model control group. Among them, the TNF-α concentration in the medium-dose group was (85.97±19.41)pg/ml, which was statistically significant compared with the model control group (P<0.05). The TNF-α concentration in the Fufang Banmao capsule (BM) group was (51.34±9.78)pg/ml, also statistically significant compared with the model control group (P<0.05). Moreover, the medium-dose PRGXP group showed a statistically significant difference compared with the BM group (P<0.05).

④ The concentrations of IFN-γ in the high- and medium-dose PRGXP groups were both higher than those in the model control group. Specifically, the IFN-γ concentration in the medium-dose group was (90.85±11.91)pg/ml, which was statistically significant compared with the model control group (P<0.05). The IFN-γ concentration in the Fufang Banmao capsule (BM) group was (66.73±11.76)pg/ml, also statistically significant compared with the model control group (P<0.05). Furthermore, the medium-dose PRGXP group showed a statistically significant difference compared with the BM group (P<0.05).

Conclusion: Peishiruanganxiaopiwan has an inhibitory effect on tumors, can increase the indices of the thymus and spleen in H₂₂ tumor-bearing mice, enhance the secretion of TNF-α and IFN-γ, boost the activity of cytokines, improve the body’s immune function, thereby exerting an antitumor effect.

Keywords: Peishiruanganxiaopiwan; H₂₂ (liver cancer); TNF-α; IFN-γ; immune function

ABSTRACT

Objective: After the observation of Peishiruanganxiaopiwan (PRGXP) on the serum TNF-α and IFN-γ in H₂₂ Tumor-bearing Mice, we have discussed the regulating and antitumor effects of PRGXP on the immune system of Tumor-bearing Mice.

Methods: Firstly, we established the tumor-bearing mice model by hypodermic inoculation in the right front axilla of mice. All the mice were divided into blank group, model group, great dose group of PRGXP, middle dose group of PRGXP, little dose group of PRGXP and FufangBanmao capsule (BM) group. Every group was dosed for ten days. After 24h of the last dose, the eyeballs of the mice were extirpated for blood collection with clean test tube. The blood was solidified for 2 hours at room temperature, and then centrifuged 1500 rpm for 10 minutes. The serum was collected, packaged and saved at −20°C. And then we measured the density of the TNF-α and IFN-γ in H₂₂ Tumor-bearing Mice using the ELISA method. Lastly, we observed regulating effects of PRGXP on the tumor.

Results:

1. According to the dosage of the PRGXP, three groups can be divided: the big, the middle and the small. The weight of tumor in the three groups separately are (0.926±0.237)g, (0.776±0.122)g and (0.935±0.227)g, which lower than the model group, corresponding to the antitumor rate are 24.6%, 36.8% and 23.9%.

2. The TI and SI of the three groups are higher than the model group. The TI of the middle group is (53.4±15.6)mg/10g, the percentage increase is 31.4% than the model group. The SI of the middle group is (65.0±10.1)mg/10g. The percentage increase is 45.1% than the model group.

3. The density of the TNF-α in three group are higher than the model group, out of which, the TNF-α density of the middle group is (85.97±19.41)pg·ml⁻¹. The middle group has statistic significance in compare with the model group (p<0.05). The density of the TNF-α in the FufangBanmao capsule (BM) group is (51.34±9.78)pg·ml⁻¹. The middle group has statistic significance in compare with the model group (p<0.05). The PRGXP middle group has statistic significance in compare with the BM group (p<0.05).

4. The density of IFN-γ in the two groups is higher than the model group, out of which, the IFN-γ density of the middle group is (90.85±11.91)pg·ml⁻¹. The middle group has statistic significance in compare with the model group (p<0.05). The density of the IFN-γ in the FufangBanmao capsule (BM) group is (66.73±11.76)pg·ml⁻¹. The PRGXP group also has statistic significance in compare with the Fu

裴正学系列方药的研究

fangBanmao capsule(BM) group (p<0.05).

conclusion: Peishiruanganxiaopiwan (PRGXP) can control tumor growth, increase the weight of thymus and spleen in H22 Tumor—bearing Mice, enhance the secretion of TNF—α and IFN—γ, boost up the activity of cells and improve the immunologic function of the body. All of these can play the roles of antitumor.

key words: Peishiruanganxiaopiwan; H22 (transplanted liver cancer); TNF—α; IFN—γ; immunologic function

Foreword

Primary hepatocellular carcinoma (PHC), commonly referred to as liver cancer, is one of the most common malignant tumors. It ranks third among malignant tumor-related deaths in China, second only to gastric cancer and esophageal cancer [1], and is particularly prevalent among middle-aged men. Due to its high malignancy, rapid disease progression, and insidious nature, it is generally difficult to detect in the early stages. Most liver cancer patients seek medical attention only when the disease has already progressed to the middle or late stages, making surgical resection unsuitable. Even for patients who are eligible for surgery, the two-year recurrence rate remains as high as 50% [2]. There is currently no standard treatment protocol for advanced liver cancer, resulting in a poor prognosis. Therefore, there is an urgent need to find more effective therapeutic approaches. At present, primary liver cancer has been listed as one of the key diseases for cancer prevention and control in China under the "Outline of China's Cancer Prevention and Control Plan" [3].

Currently, the mainstay of liver cancer treatment in the early stages is surgical resection, supplemented by radiotherapy and chemotherapy. However, according to existing data, the radical cure rate of surgery is relatively low, and the postoperative recurrence rate is high. In addition, liver cancer is often insensitive to radiotherapy and chemotherapy and prone to metastasis. As a result, most patients are diagnosed at an advanced stage, rendering surgery ineffective. While the medical community continues to search for and use anticancer drugs, it has also discovered that these drugs often come with significant toxic side effects: many chemical anticancer agents, while targeting cancer cells, tend to affect normal cells as well, and may cause varying degrees of mutagenic and genotoxic effects. Consequently, treating tumors can sometimes increase the risk of patients developing secondary cancers. In contrast, plant-based medicines appear to have less obvious genotoxicity [4–5]. Traditional Chinese medicine, on the other hand, offers unique advantages in alleviating clinical symptoms, improving quality of life, and prolonging survival. Therefore, finding a comprehensive and clinically effective treatment plan for liver cancer is an important means of extending patients' survival time and improving their quality of life, and holds tremendous practical significance.

Primary liver cancer falls under categories such as "liver accumulation," "fat qi," "accumulation," and "distension" in traditional Chinese medicine. The understanding of its etiology and pathogenesis is that it arises from deficiency of vital energy. As stated in the "Suwen: On Acupuncture Techniques": "When vital energy is intact, evil cannot invade; where evil gathers, it must be due to deficiency of vital energy." Similarly, the "Yizong Bidu" notes: "Accumulation occurs because vital energy is insufficient, allowing pathogenic factors to take hold." Thus, traditional Chinese medicine theory holds that deficiency of vital energy is the primary cause of cancer. As Zhang Yuan put it: "Healthy people do not develop accumulations; only those with weak vital energy do."

Peishiruanganxiaopiwan is a specialized formula developed over forty years by Professor Pei Zhengxue, a nationally renowned expert in integrated Chinese and Western medicine, for treating primary liver cancer and gastric cancer. It has demonstrated remarkable clinical efficacy. Several cases of primary liver cancer and gastric cancer have been completely cured using this formula, and it has also shown varying degrees of clinical effectiveness in treating most patients who are not suitable for surgery. Professor Pei believes that primary liver cancer results from deficiency of vital energy, dysfunction of internal organs, and stagnation of qi, blood stasis, and phlegm accumulation. This condition is characterized by underlying deficiency and overt excess—deficiency of vital energy combined with invasion of pathogenic factors. Based on this understanding, he concluded that the fundamental cause of malignant tumor occurrence and development is "deficiency of vital energy," and the basic principle for treating malignant tumors is to strengthen vital energy and consolidate the root. Accordingly, he proposed the concept of "strengthening vital energy to eliminate accumulation, eliminating accumulation to consolidate the root," and subsequently formulated the "Liver Cancer Formula." Through continuous clinical practice, refinement, and reorganization, this formula eventually evolved into "Peishiruanganxiaopiwan" (composed of Bupleurum, raw turtle shell, pangolin scales, soapberry spines, Citrus aurantium, Hedyotis diffusa, Scutellaria barbata, Sparganium stoloniferum, Curcuma wenyujin, white peony, seaweed, kelp, Salvia miltiorrhiza, Astragalus membranaceus, and other ingredients). Over many years

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裴正学系列方药的研究

clinical application has proven that this medicine has excellent effects in treating primary liver cancer, preventing and controlling recurrence and metastasis, improving patients' weakened immune function caused by radiotherapy and chemotherapy, enhancing their quality of life, and prolonging their survival. This experiment aims to scientifically and objectively study and demonstrate the antitumor effects and mechanisms of Peishiruanganxiaopiwan, to systematically organize the valuable clinical experience of the mentor, and to provide sufficient scientific theoretical basis for its widespread clinical application.

Rationale for the Study