Chinese Abstract

Chinese Abstract

Objective: By observing the effects of Pei’s Soft Liver and Relieve Distention Pill (PRGXP) on the expression of P27 and Bcl-2 in H22 transplanted liver cancer tissue in mice, this study aims to explore how PRGXP regulates the immune system of tumor-bearing mice and inhibits tumors, thereby analyzing its mechanism of action in treating primary liver cancer and providing a theoretical basis for its clinical application.

Methods: A mouse model of H22 (liver cancer) solid tumor was established by subcutaneous inoculation in the right anterior axilla. The mice were randomly divided into a blank group, a model control group, a low-dose PRGXP group, a medium-dose PRGXP group, a high-dose PRGXP group, and a Fufang Banmao Capsule (BM) group. After 10 days of gavage administration, the mice were euthanized by spinal cord dislocation, and the tumors, thymuses, and spleens were excised and weighed. The tumor inhibition rate, thymus index (TI), and spleen index (SI) were calculated. The expression levels of P27 and Bcl-2 proteins in the H22 transplanted liver cancer tissue were determined using immunohistochemistry.

Results:

1. The average tumor weights in the low-, medium-, and high-dose PRGXP groups were all lower than those in the model control group, at (0.935±0.227)g, (0.776±0.122)g, and (0.926±0.237)g, respectively, with statistically significant differences compared with the model control group (p<0.05). The corresponding tumor inhibition rates were 23.9%, 36.8%, and 24.5%.
2. The thymus index (TI) and spleen index (SI) in the low-, medium-, and high-dose PRGXP groups were all higher than those in the model control group, with the medium-dose group showing a TI of (53.2±15.6) mg/10g, a 31.4% increase compared with the model control group, which was statistically significant (p<0.05). The SI values for the low-, medium-, and high-dose PRGXP groups were (58.9±14.5) mg/10g, (65.0±10.1) mg/10g, and (55.2±15.9) mg/10g, respectively, all showing statistically significant differences compared with the model control group (p<0.05).
3. The positive expression of P27 in the low-, medium-, and high-dose PRGXP groups was stronger than that in the model control group, with statistically significant differences compared with the model control group (p<0.05).
4. The positive expression of Bcl-2 in the low-, medium-, and high-dose PRGXP groups was weaker than that in the model control group, with statistically significant differences compared with the model control group (p<0.05). Notably, the medium-dose PRGXP group had lower Bcl-2 expression in the tumor tissue than the BM group, which was also statistically significant (p<0.05).

Conclusion: Pei’s Soft Liver and Relieve Distention Pill has the effect of inhibiting tumor growth, can increase the thymus index and spleen index in H22 tumor-bearing mice, and exerts an antitumor effect by increasing the expression of P27 protein while reducing the expression of Bcl-2 protein.

Keywords: Pei’s Soft Liver and Relieve Distention Pill; H22 (liver cancer); P27; Bcl-2

Research on Pei Zhengxue’s Series of Formulas and Medicines

ABSTRACT

Objective: Through observing the effect of Peishiruanganxiaopi pills (PRGXP) on the P27 and Bcl-2 in H22 tumor-bearing mice, we have discussed the regulating and antitumor effects of PRGXP on the immune system of tumor-bearing mice. To support for its clinical use, we aim to explore the basic mechanism of PRGXP's effect on primary carcinoma of the liver.

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, PRGXP low-dose group, PRGXP medium-dose group, PRGXP high-dose group, and Fufang Banmao capsule (BM) group. Ten days later, they were fed and then killed. Those tumors, thymuses and spleens were peeled off and weighted. The antitumor rate, the thymus index (TI) and the spleen index (SI) were calculated. By studying the expression of P27 and Bcl-2 protein in the tumor tissue by immunohistochemistry, we analyzed the statistics.

Results:

1. According to the dosage of the PRGXP, three groups can be divided: the low, the medium and the high. The weight of tumor in the three groups of PRGXP, separately is (0.935±0.227)g, (0.776±0.122)g and (0.926±0.237)g, which is lower than the model group (p<0.05) while the antitumor rate are 23.9%, 36.8% and 24.5%.
2. The TI and SI of the three groups are higher than the model group (p<0.05). The TI of the PRGXP medium-dose group is (53.2±15.6) mg/10g, and the percentage increase is 31.4% than the model group (p<0.05). The SI in low-dose group, medium-dose group and high-dose group of the PRGXP were (58.9±14.5) mg/10g, (65.0±10.1) mg/10g, (55.2±15.9) mg/10g, which indicated that the three groups were all statistical significances compared with model group (p<0.05).
3. The expression of P27 was increased in PRGXP groups and BM group, which were all statistical significances compared with model group (p<0.05).
4. The expression of Bcl-2 was decreased in PRGXP groups and BM group, which were all statistical significances compared with model group (p<0.05). The middle dose group of PRGXP has significant antitumor effect.

Conclusion: PRGXP could inhibit the growth of liver cancer, and could significantly increase the thymus index and the spleen index in H22 tumor-bearing. PRGXP was also shown significant antitumor effect through the increased expression of P27 and the expressions of Bcl-2 reduced.

Keywords: Peishiruanganxiaopi pill; H22 (transplanted liver cancer); P27; Bcl-2

Preface

Primary hepatocellular carcinoma (referred to as liver cancer) is a common malignant tumor clinically occurring in bile duct cells and hepatocytes. According to the "WHO 2008"

Research on Pei Zhengxue’s Series of Formulas and Medicines

The "World Cancer Report" lists primary hepatocellular carcinoma as one of the top ten malignant tumors, ranking fifth globally and third in mortality. The incidence of liver cancer has been increasing year by year worldwide, with approximately 620,000 new cases annually, of which 55% occur in China, making it the second leading cause of cancer-related deaths in our country [1][2]. Liver cancer has an insidious onset, with inconspicuous early symptoms, rapid disease progression, high malignancy, and a tendency to recur, resulting in a low five-year survival rate and posing a serious threat to human health. Liver cancer has a variety of treatment options, including surgery, chemotherapy, radiotherapy, minimally invasive therapy, biological therapy, and orthotopic liver transplantation. Surgery is the preferred treatment for early-stage liver cancer; however, most patients lose the opportunity for surgery by the time they seek medical attention. Additionally, due to low tolerance to radiotherapy, severe adverse reactions after chemotherapy, and poor sensitivity of liver cancer to chemotherapeutic drugs, the therapeutic outcomes are often unsatisfactory. Traditional Chinese medicine, a distinctive feature of China's healthcare system, has achieved good results in treating liver cancer. In the early stages of liver cancer, TCM treatment can improve patients' tolerance to surgery; postoperative use can strengthen their physical condition and eliminate residual lesions; for patients undergoing radiotherapy or chemotherapy, it can alleviate toxic side effects and enhance therapeutic efficacy; long-term use of TCM can effectively prevent recurrence and metastasis of liver cancer, improve patients' quality of life, and prolong survival. Therefore, comprehensive prevention and treatment of liver cancer primarily based on TCM is increasingly gaining attention.

Based on clinical manifestations such as hepatomegaly, pain in the hepatic region, jaundice, and signs of cirrhosis, the TCM disease names for liver cancer fall under categories like "accumulation," "liver accumulation," "jaundice," and "distention." According to "The Treatise on Causes and Symptoms of Various Diseases," "Piqi is a condition where liver disease spreads to the spleen, and the spleen should then transmit the pathogenic factor to the kidneys. However, since the kidneys are strongest in winter and cannot accept external pathogens, while the spleen wishes to return to the liver but the liver refuses, the pathogenic factor accumulates and forms an accumulation." Furthermore, "Jisheng Fang·Tumor Accumulation Chapter" states, "The manifestation of fatty qi is a mass in the left hypochondrium, about the size of an overturned cup, large and with a head and tail-like appearance—this is liver accumulation." The main TCM etiology and pathogenesis of liver cancer involve deficiency of vital energy and stagnation of pathogenic factors, thus classifying liver cancer as a syndrome of deficiency of vital energy and excess of pathogenic factors. As stated in "Essential Readings on Medical Principles," "Accumulation arises because vital energy is deficient, allowing pathogenic factors to take hold." The primary TCM therapeutic principles for liver cancer include strengthening the spleen and benefiting qi, clearing heat and detoxifying, tonifying the liver and kidneys, promoting blood circulation and removing blood stasis, and transforming phlegm and eliminating dampness.

Pei's Soft Liver and Qi-Relieving Pill is an effective prescription for treating liver cancer, summarized by Professor Pei Zhengxue, a renowned expert in integrated traditional Chinese and Western medicine, through more than forty years of clinical practice. The formula mainly consists of Salvia miltiorrhiza, Astragalus membranaceus, Bupleurum chinense, Paeonia lactiflora, Curcuma wenyujin, Laminaria japonica, Ziziphus jujuba spines, Hedyotis diffusa, and Scutellaria barbata, among others. It has the effects of soothing the liver and regulating qi, softening hard masses and dispersing nodules, breaking blood stasis and removing blood clots, clearing heat and detoxifying, reinforcing vital energy while expelling pathogenic factors, and addressing both the root cause and symptoms. Pei's Soft Liver and Qi-Relieving Pill has achieved satisfactory therapeutic effects in treating primary liver cancer and gastric cancer, particularly in alleviating adverse reactions caused by radiotherapy and chemotherapy and improving patients' quality of life.

This experiment employs animal studies, using immunohistochemistry and image analysis to calculate and analyze the tumor inhibition rate, thymus index, and spleen index in H22 tumor-bearing mice treated with Pei's Soft Liver and Qi-Relieving Pill. It also examines the expression levels of P27 and Bcl-2 from the perspectives of cell proliferation and apoptosis, aiming to investigate the mechanism of tumor suppression by Pei's Soft Liver and Qi-Relieving Pill and provide a theoretical basis for its widespread clinical application.

Rationale for the Study