A traditional Chinese medicine composition for preventing and treating recurrence of liver cancer after operation

Abstract

The application provides a traditional Chinese medicine composition for preventing and treating recurrence of liver cancer after operation, and relates to the technical field of biological medicine. The patient after operation of liver cancer is mainly characterized by deficiency of healthy qi, uncleaned residual toxin and mutual knotting of blood stasis and heat, the application takes strengthening healthy qi and tonifying the body, detoxifying and removing blood stasis and regulating qi movement as the main therapeutic principle, and is matched with the traditional Chinese medicine composition composed of caragana, red ginseng, giant knotweed, radix curdlariae, lantana camara, eurypalum odoratum, kudou, magnolia flower and licorice root, and the application of the traditional Chinese medicine composition in preparing the medicine for preventing and treating recurrence of liver cancer after operation is provided. The whole prescription can strengthen healthy qi and tonify the body without leaving evil, expel evil without damaging healthy qi, and the combined drugs can jointly play the effects of invigorating the spleen and kidney, detoxifying and removing blood stasis and promoting qi and activating blood, and are suitable for the syndrome of deficiency of healthy qi and blood stasis of residual toxin after operation of liver cancer, and can effectively reduce the risk of recurrence of liver cancer after operation.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to a traditional Chinese medicine composition for preventing and treating postoperative recurrence of liver cancer. Background Technology

[0002] Primary liver cancer (PLC) is one of the most common malignant tumors worldwide and one of the leading causes of death from malignant tumors, with hepatocellular carcinoma (HCC) accounting for approximately 95%. Despite significant advancements in the diagnosis and treatment of HCC in recent years, surgical resection remains one of the most effective treatments for early-stage liver cancer. However, the 5-year recurrence rate after surgery is as high as 70%, severely impacting treatment outcomes and patients' quality of life. Furthermore, there is currently a lack of standard adjuvant therapy regimens for early-stage liver cancer recurrence and metastasis after surgery.

[0003] Currently, Western medicine treats recurrent liver cancer using methods such as re-hepatectomy, salvage liver transplantation, radiofrequency ablation combined with sorafenib, and targeted therapies like regorafenib and ramucirumab. These methods are expensive and often have limited efficacy. Traditional Chinese medicine (TCM) offers advantages over Western medicine, including synergistic effects, reduced toxicity, and prolonged survival. TCM plays a vital role in alleviating clinical symptoms and reducing the risk of liver cancer occurrence and recurrence. The integration of TCM and Western medicine has become an important principle in the multidisciplinary treatment system for liver cancer. Summary of the Invention

[0004] (a) Technical problems to be solved

[0005] In view of the shortcomings of the prior art, the present invention provides a traditional Chinese medicine composition for preventing and treating postoperative recurrence of liver cancer.

[0006] (II) Technical Solution

[0007] To achieve the above objectives, the present invention provides the following technical solution:

[0008] This invention provides a traditional Chinese medicine composition for preventing postoperative recurrence of liver cancer. The composition is made from the following traditional Chinese medicinal materials in parts by weight: 10-30 parts of Caragana korshinskii, 1-15 parts of red ginseng, 5-20 parts of Polygonum cuspidatum, 5-20 parts of Codonopsis pilosula, 5-20 parts of Lycopus lucidus, 5-20 parts of Semiaquilegia adoxoides, 5-20 parts of Viola yedoensis, 5-20 parts of Magnolia officinalis, and 1-15 parts of prepared licorice root.

[0009] Preferably, the traditional Chinese medicine composition of the present invention for preventing postoperative recurrence of liver cancer is made from the following traditional Chinese medicinal materials in parts by weight: 15-25 parts of Caragana korshinskii, 3-10 parts of red ginseng, 10-20 parts of Polygonum cuspidatum, 10-20 parts of Codonopsis pilosula, 10-20 parts of Lycopus lucidus, 10-20 parts of Semiaquilegia adoxoides, 10-20 parts of Viola yedoensis, 10-20 parts of Magnolia officinalis, and 1-10 parts of prepared licorice root.

[0010] Preferably, the traditional Chinese medicine composition of the present invention for preventing postoperative recurrence of liver cancer is made from the following traditional Chinese medicinal materials in parts by weight: 21 parts of Caragana korshinskii, 6 parts of red ginseng, 15 parts of Polygonum cuspidatum, 12 parts of Codonopsis pilosula, 18 parts of Lycopus lucidus, 12 parts of Semiaquilegia adoxoides, 15 parts of Viola yedoensis, 15 parts of Magnolia officinalis, and 6 parts of prepared licorice root.

[0011] Furthermore, the present invention also provides the application of a traditional Chinese medicine composition made from the above-mentioned parts by weight of Chinese medicinal materials in the preparation of a drug for preventing and treating postoperative recurrence of liver cancer.

[0012] Specifically, the drug is made from the above-mentioned proportions of Chinese medicinal materials by weight, prepared according to conventional methods, and supplemented with pharmaceutically acceptable excipients, into any one of the following oral dosage forms: granules, formulated granules, oral liquids, capsules, tablets, pills, and decoctions.

[0013] All medicinal materials in the herbal composition of this invention are selected from those processed according to relevant regulations. The source and efficacy of each medicinal material are described below:

[0014] Caragana: The flower of Caragana, a plant in the genus Caragana of the legume family. It is sweet and slightly warm in nature, and enters the spleen and kidney meridians. It has the effects of strengthening the spleen and kidney, harmonizing blood and dispelling wind, and detoxifying. It is mainly used to treat consumptive cough, dizziness and tinnitus, soreness and weakness of the waist and knees, qi deficiency, poor eruption of smallpox, gout, and traumatic injuries.

[0015] Red ginseng: The dried root and rhizome of Panax ginseng CAMey., a plant of the Araliaceae family, after steaming. It has the effects of greatly replenishing vital energy, restoring pulse and consolidating the body, and benefiting qi and controlling bleeding. It is suitable for those with physical weakness and impending collapse, cold limbs and weak pulse, qi failing to control blood, and metrorrhagia.

[0016] Polygonum cuspidatum: The dried rhizome and root of Polygonum cuspidatum Sieb. et Zucc., a plant belonging to the Polygonaceae family. This herb has the effects of promoting diuresis and relieving jaundice, clearing heat and detoxifying, dispersing blood stasis and relieving pain, and relieving cough and resolving phlegm. It is mainly used to treat damp-heat jaundice, urinary tract infections, leukorrhea, rheumatic pain, carbuncles and boils, burns, amenorrhea, abdominal masses, traumatic injuries, and cough due to lung heat.

[0017] Codonopsis pilosula: The dried root of Codonopsis pilosula Wolff, a plant of the genus Codonopsis in the family Apiaceae; it has the effects of moistening the lungs and resolving phlegm, nourishing yin and stomach, calming the liver and detoxifying; it is often used to treat lung heat cough, vomiting and nausea, poor appetite and dry mouth, red eyes and dizziness, boils and carbuncles and other diseases.

[0018] Ze Lan: The dried aerial parts of Lycopus lucidus Turcz. var. hirtus Regel, a plant belonging to the genus Lycopus in the Lamiaceae family; it has the effects of promoting blood circulation and regulating menstruation, removing blood stasis and eliminating carbuncles, and promoting diuresis and reducing swelling; it is mainly used to treat irregular menstruation, amenorrhea, dysmenorrhea, postpartum abdominal pain due to blood stasis, carbuncles and boils, and edema and ascites.

[0019] Seminiaquilegia adoxoides (DC.) Makino, a plant in the Ranunculaceae family, is a dried tuberous root. It has the effects of clearing heat and detoxifying, reducing swelling and dissipating nodules. It is used for carbuncles, boils, mastitis, scrofula, and snake and insect bites.

[0020] Corydalis bungeana Turcz., a plant belonging to the genus Corydalis in the Papaveraceae family, is a dried whole herb. It has the effects of clearing heat and detoxifying, dispersing nodules and reducing swelling; it is mainly used to treat epidemic colds, sore throat, boils and carbuncles, mumps, and erysipelas.

[0021] Magnolia officinalis flower: The dried flower buds of Magnolia officinalis Rehd. et Wils. or Magnolia officinalis Rehd. et Wils. var. biloba Rehd. et Wils., belonging to the genus Magnolia of the family Magnoliaceae; it has the effects of aromatically resolving dampness, regulating qi, and relieving chest tightness. It is mainly used to treat dampness obstruction and qi stagnation in the spleen and stomach, chest and epigastric fullness and distension, and poor appetite.

[0022] Prepared licorice root: A processed form of licorice root in traditional Chinese medicine, it is the dried root and rhizome of Glycyrrhiza uralensis Fisch., Glycyrrhiza inflata Bat., or Glycyrrhiza glabra L., all belonging to the legume family. It is prepared through cleaning, cutting, and honey-processing. Its main functions are to tonify the spleen and stomach, replenish qi, and restore pulse. It is commonly used to treat symptoms such as spleen and stomach weakness, fatigue, insufficient heart qi, palpitations, and irregular pulse.

[0023] (III) Beneficial Effects

[0024] Based on extensive literature review and clinical experience, postoperative liver cancer patients often present with a core pathogenesis of deficiency of vital energy, residual toxins, and the accumulation of blood stasis and heat. Therefore, this invention adopts the main treatment principles of strengthening the body's resistance, detoxifying and resolving blood stasis, and regulating Qi flow. It incorporates a traditional Chinese medicine composition consisting of Caragana korshinskii, red ginseng, Polygonum cuspidatum, Codonopsis pilosula, Lycopus lucidus, Semiaquilegia adoxoides, Viola yedoensis, Magnolia officinalis flower, and prepared licorice root. The invention also provides the application of this composition in the preparation of drugs for preventing postoperative recurrence of liver cancer. In this invention, Caragana korshinskii is the principal ingredient. Caragana korshinskii is sweet and warm, entering the spleen and kidney meridians. The spleen is the foundation of acquired constitution, and the kidney is the foundation of innate constitution. Its spleen-strengthening and kidney-tonifying effects support vital energy and enhance the body's anti-cancer ability; its blood-regulating, wind-dispelling, and detoxifying effects can also clear residual cancer toxins after surgery. Therefore, it addresses both the root cause and the symptoms, adhering to the principle of simultaneously strengthening the body's resistance and eliminating pathogenic factors. Polygonum cuspidatum clears heat and detoxifies, disperses blood stasis and relieves pain, and also promotes diuresis and reduces jaundice; Semiaquilegia adoxoides clears heat and detoxifies, reduces swelling and dissipates nodules; Viola yedoensis clears heat and detoxifies, dissipates nodules and eliminates masses; the three herbs are cold in nature and bitter in taste, which can clear the residual heat and toxins after surgery, promote blood circulation and dissipate nodules to inhibit residual lesions, and together they serve as assistant herbs to the principal herbs to attack pathogens. Red ginseng greatly replenishes vital energy and nourishes blood, assisting the principal herb in strengthening the body's resistance and consolidating its foundation. It also counteracts the harmful effects of bitter and cold herbs like Polygonum cuspidatum, which can damage the body's vital energy. Furthermore, its warm nature complements the principal herb, providing warmth without causing dryness. Eupatorium fortunei invigorates blood circulation, removes blood stasis, promotes diuresis, and reduces swelling, assisting the principal herb in dispersing nodules and detoxifying, clearing stasis and unblocking the meridians, and eliminating residual stagnation in the liver meridians. Codonopsis pilosula nourishes yin and harmonizes the stomach, moistens the lungs and resolves phlegm, and clears and protects the liver. It can replenish postoperative liver yin deficiency, clear liver stagnation and deficiency heat, and moisten dryness without retaining pathogens. Magnolia officinalis flower regulates qi, relieves stagnation, and resolves dampness, harmonizing the qi mechanism of the middle jiao, ensuring that tonification is not stagnant and clearing is not harmful to the stomach. These four herbs serve as adjuvant herbs. Finally, prepared licorice root is used as the guiding herb to invigorate qi and harmonize the middle jiao, harmonizing the properties of all the herbs in the formula. The combined effects of these herbs strengthen the body's resistance without retaining pathogens and eliminate pathogens without harming the body's vital energy, achieving the effects of strengthening the spleen and kidneys, detoxifying and removing blood stasis, and invigorating qi and blood. This formula is suitable for patients with deficiency and toxicity after liver cancer surgery and can effectively reduce the risk of recurrence after liver cancer surgery.

[0025] Following the principles of traditional Chinese medicine (TCM) formulation, in vitro evaluation of its anti-hepatocellular carcinoma activity further demonstrates that the components of this invention are rationally combined, exhibiting synergistic effects. This significantly inhibits hepatocellular carcinoma cell proliferation and promotes apoptosis, demonstrating excellent anti-hepatocellular carcinoma activity. Furthermore, in vivo experiments show that this invention's TCM composition significantly slows tumor growth in mice with recurrent hepatocellular carcinoma in situ after surgery, reducing tumor volume and weight, exhibiting excellent in vivo tumor-suppressing effects. Simultaneously, compared to 5-FU, this invention's TCM composition better improves the survival status of mice with recurrent hepatocellular carcinoma in situ after surgery. Additionally, this invention's TCM composition can promote CD4+... + T lymphocyte levels, inhibiting CD8 + By regulating T lymphocyte levels, the immune function can be improved, immune imbalance in relapsed mice can be improved, and immune function can be strengthened after liver cancer surgery, thereby achieving the goal of preventing liver cancer recurrence. Attached Figure Description

[0026] Figure 1Cell viability (a) and apoptosis rate (b) of HepG2 cells after treatment with different drug-containing serums; compared with the control group, **P<0.01; compared with drug 1, # P<0.05, ## P<0.01.

[0027] Figure 2 Tumor growth curve (a) and tumor quality (b); **P<0.01 compared with the model group.

[0028] Figure 3 CD4 in peripheral blood (ac) and spleen (df) + CD8 + T lymphocyte subset expression and CD4 + / CD8 + Ratio; compared with the model group, **P<0.01. Detailed Implementation

[0029] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0030] Example 1

[0031] A drug for preventing recurrence after liver cancer surgery, the drug being a traditional Chinese medicine composition consisting of the following parts by weight of Chinese medicinal materials: 10 parts of Caragana korshinskii, 1 part of red ginseng, 5 parts of Polygonum cuspidatum, 5 parts of Codonopsis pilosula, 5 parts of Lycopus lucidus, 5 parts of Semiaquilegia adoxoides, 5 parts of Viola yedoensis, 5 parts of Magnolia officinalis, and 1 part of Glycyrrhiza uralensis (processed), which are prepared according to conventional methods, supplemented with pharmaceutically acceptable excipients, and made into any one of the following oral dosage forms: granules, formulated granules, oral liquid, capsules, tablets, pills, or decoction.

[0032] Example 2

[0033] A drug for preventing recurrence after liver cancer surgery, the drug being a traditional Chinese medicine composition consisting of the following parts by weight of Chinese medicinal materials: 30 parts of Caragana korshinskii, 15 parts of red ginseng, 20 parts of Polygonum cuspidatum, 20 parts of Codonopsis pilosula, 20 parts of Lycopus lucidus, 20 parts of Semiaquilegia adoxoides, 20 parts of Viola yedoensis, 20 parts of Magnolia officinalis flower, and 15 parts of prepared licorice root, which are prepared according to conventional methods and supplemented with pharmaceutically acceptable excipients to produce any one of the following oral dosage forms: granules, formulated granules, oral liquid, capsules, tablets, pills, or decoction.

[0034] Example 3

[0035] A drug for preventing postoperative recurrence of liver cancer, comprising a traditional Chinese medicine composition in the following parts by weight: 14 parts of *Caragana korshinskii*, 2 parts of red ginseng, 8 parts of *Polygonum cuspidatum*, 9 parts of *Codonopsis pilosula*, 9 parts of *Lycopus lucidus*, 6 parts of *Semiaquilegia adoxoides*, 6 parts of *Viola yedoensis*, 6 parts of *Magnolia officinalis*, and 2 parts of prepared licorice root. These are prepared using conventional methods, supplemented with pharmaceutically acceptable excipients, and formulated into any one of the following oral dosage forms: granules, formulated granules, oral liquid, capsules, tablets, pills, or decoction.

[0036] Example 4

[0037] A drug for preventing recurrence after liver cancer surgery, the drug being a traditional Chinese medicine composition consisting of the following parts by weight of Chinese medicinal materials: 15 parts of Caragana korshinskii, 3 parts of red ginseng, 10 parts of Polygonum cuspidatum, 10 parts of Codonopsis pilosula, 10 parts of Lycopus lucidus, 10 parts of Semiaquilegia adoxoides, 10 parts of Viola yedoensis, 10 parts of Magnolia officinalis, and 3 parts of Glycyrrhiza uralensis (processed), which are prepared according to conventional methods, supplemented with pharmaceutically acceptable excipients, and made into any one of the following oral dosage forms: granules, formulated granules, oral liquid, capsules, tablets, pills, or decoction.

[0038] Example 5

[0039] A drug for preventing postoperative recurrence of liver cancer, the drug being a traditional Chinese medicine composition consisting of the following parts by weight of Chinese medicinal materials: 25 parts of Caragana korshinskii, 8 parts of red ginseng, 20 parts of Polygonum cuspidatum, 20 parts of Codonopsis pilosula, 20 parts of Lycopus lucidus, 20 parts of Semiaquilegia adoxoides, 20 parts of Viola yedoensis, 20 parts of Magnolia officinalis, and 6 parts of Glycyrrhiza uralensis (processed), which are prepared according to conventional methods, supplemented with pharmaceutically acceptable excipients, and made into any one of the following oral dosage forms: granules, formulated granules, oral liquid, capsules, tablets, pills, or decoction.

[0040] Example 6

[0041] A drug for preventing recurrence after liver cancer surgery, the drug being a traditional Chinese medicine composition consisting of the following parts by weight of Chinese medicinal materials: 18 parts of Caragana korshinskii, 5 parts of red ginseng, 12 parts of Polygonum cuspidatum, 10 parts of Codonopsis pilosula, 12 parts of Lycopus lucidus, 10 parts of Semiaquilegia adoxoides, 12 parts of Viola yedoensis, 12 parts of Magnolia officinalis, and 3 parts of Glycyrrhiza uralensis (processed), which are prepared according to conventional methods, supplemented with pharmaceutically acceptable excipients, and made into any one of the following oral dosage forms: granules, formulated granules, oral liquid, capsules, tablets, pills, or decoction.

[0042] Example 7

[0043] A drug for preventing recurrence after liver cancer surgery, the drug being a traditional Chinese medicine composition consisting of the following parts by weight of Chinese medicinal materials: 24 parts of Caragana korshinskii, 9 parts of red ginseng, 18 parts of Polygonum cuspidatum, 15 parts of Codonopsis pilosula, 15 parts of Lycopus lucidus, 15 parts of Semiaquilegia adoxoides, 18 parts of Viola yedoensis, 18 parts of Magnolia officinalis, and 8 parts of Glycyrrhiza uralensis (processed), which are prepared according to conventional methods, supplemented with pharmaceutically acceptable excipients, and made into any one of the following oral dosage forms: granules, formulated granules, oral liquid, capsules, tablets, pills, or decoction.

[0044] Example 8

[0045] A drug for preventing recurrence after liver cancer surgery, the drug being a traditional Chinese medicine composition consisting of the following parts by weight of Chinese medicinal materials: 21 parts of Caragana korshinskii, 6 parts of red ginseng, 15 parts of Polygonum cuspidatum, 12 parts of Codonopsis pilosula, 18 parts of Lycopus lucidus, 12 parts of Semiaquilegia adoxoides, 15 parts of Viola yedoensis, 15 parts of Magnolia officinalis, and 6 parts of Glycyrrhiza uralensis (processed), which are prepared according to conventional methods, supplemented with pharmaceutically acceptable excipients, and made into any one of the following oral dosage forms: granules, formulated granules, oral liquid, capsules, tablets, pills, or decoction.

[0046] Experimental Example 1

[0047] In vitro anti-hepatocellular carcinoma activity evaluation

[0048] 1 Experimental Drugs

[0049] Drug 1: 21 parts Caragana korshinskii, 6 parts red ginseng, 15 parts Polygonum cuspidatum, 12 parts Codonopsis pilosula, 18 parts Lycopus lucidus, 12 parts Semiaquilegia adoxoides, 15 parts Viola yedoensis, 15 parts Magnolia officinalis, 6 parts prepared licorice root.

[0050] Drug 2: 15 parts Polygonum cuspidatum, 12 parts Codonopsis pilosula, 18 parts Lycopus lucidus, 12 parts Semiaquilegia adoxoides, 15 parts Viola yedoensis, 15 parts Magnolia officinalis, 6 parts Glycyrrhiza uralensis (processed).

[0051] Drug 3: 21 parts of Caragana korshinskii, 6 parts of red ginseng, 15 parts of Hedyotis diffusa, 12 parts of Adenophora stricta, 18 parts of Lycopus lucidus, 12 parts of Semiaquilegia adoxoides, 15 parts of Viola yedoensis, 15 parts of Magnolia officinalis flower, and 6 parts of prepared licorice root.

[0052] Drug 4: 21 parts Caragana korshinskii, 6 parts red ginseng, 15 parts Polygonum cuspidatum, 12 parts Codonopsis pilosula, 18 parts Lycopus lucidus, 12 parts Cremastra appendiculata, 15 parts Forsythia suspensa, 15 parts Magnolia officinalis flower, 6 parts prepared licorice root.

[0053] Drug 5: 21 parts Caragana korshinskii, 6 parts red ginseng, 15 parts Polygonum cuspidatum, 12 parts Codonopsis pilosula, 18 parts Verbena officinalis, 15 parts Magnolia officinalis flower, and 6 parts prepared licorice root.

[0054] Weigh each ingredient according to the above formula, and add 8 times the weight of purified water for each 0.5g portion. Boil for 40 minutes, take the decoction, add 6 times the amount of purified water to the dregs, and boil again for 30 minutes. Then, combine the two decoctions and concentrate the decoction to one-third of the total volume of the medicine. Refrigerate for later use.

[0055] 2. Preparation of drug-containing serum

[0056] Eighteen healthy SPF-grade 6-7 week old SD rats were randomly selected and, after 7 days of acclimatization, were randomly divided into a control group and drug groups 1-5, with 3 rats in each group. Rats in drug groups 1-5 were administered the corresponding drug solution by gavage, with a gavage volume of 2 mL each time. The control group was administered an equal dose of purified water by gavage. Gavage was performed once daily for 7 consecutive days. Two hours after the last gavage, rats were anesthetized with sodium pentobarbital, and blood was collected from the abdominal aorta. After centrifugation, the supernatant was collected, and the serum from rats in the same group was collected, sterilized, filtered through a 0.22 μm microporous membrane, and stored below -18℃ for later use.

[0057] 3 Effects of different drug-containing sera on the proliferation activity of HepG2 cells

[0058] After selecting HepG2 cells in the logarithmic growth phase, they were administered at a rate of 1×10⁻⁶. 5 Cells were seeded per well in 96-well plates. After 24 hours of culture and full cell adhesion, the culture medium was discarded. 80 μL of blank serum or corresponding drug-containing serum was added to the control group and drug groups 1-5, respectively. The volume was then increased to 200 μL with basal culture medium and incubated for 48 hours. Subsequently, 10 μL of CCK-8 solution was added to each well, and the plates were incubated at 37°C for 4 hours. Three replicates were set up for each group, and the absorbance of each well was measured using a microplate reader to calculate the cell viability for each group.

[0059] 4. Effects of different drug-containing sera on HepG2 cell apoptosis

[0060] Cell apoptosis was detected using the Annexin V-FITC apoptosis detection kit. Cells in logarithmic growth phase (2 × 10⁶ cells) were used. 5 (Number of cells / well) were seeded into 6-well plates. When the cell confluence reached 70-80%, the culture medium was discarded. 80 μL of blank serum or corresponding drug-containing serum was added to the control group and drug groups 1-5, respectively. The volume was then increased to 200 μL with basal culture medium. The intervention was carried out for 48 h, followed by trypsin digestion and centrifugation to collect the cells. The cells were resuspended in 5 μL Annexin V-FITC, 5 μL LPI, and 500 μL B inding buffer. The cells were incubated in the dark for 20 minutes. Apoptotic cells were detected by FACS, and the apoptosis rate was calculated using FlowJo software. Three replicates were set up for each group.

[0061] 5. Statistical Analysis

[0062] The measurement data was collected using GraphPad Prism 8.0 software. This indicates that one-way ANOVA was used for comparisons among multiple groups, and P < 0.05 was considered statistically significant.

[0063] 6 Results

[0064] like Figure 1 The results showed that, compared with blank serum, the cell viability and apoptosis rate of HepG2 cells treated with serum containing drugs 1-5 were significantly decreased (P<0.01). Specifically, the cell viability of HepG2 cells treated with serum containing drug 1 was significantly lower than that of the serum treatment group containing drugs 2-5 (P<0.05, P<0.01), while the apoptosis rate was significantly higher in the latter group (P<0.05, P<0.01). These results indicate that drugs 1-5 all exhibited varying degrees of in vitro anti-hepatocellular carcinoma activity, with drug 1 showing significantly higher anti-hepatocellular carcinoma activity than drugs 2-5.

[0065] Based on extensive prior literature review and clinical experience, this invention scientifically formulates a traditional Chinese medicine composition consisting of Caragana korshinskii, red ginseng, Polygonum cuspidatum, Codonopsis pilosula, Lycopus lucidus, Semiaquilegia adoxoides, Viola yedoensis, Magnolia officinalis flower, and prepared licorice root. Following the principles of traditional Chinese medicine's principal-assistant-adjuvant-guide formulation, the aforementioned in vitro anti-hepatocellular carcinoma activity evaluation further demonstrates that the components of this invention are rationally combined, with synergistic effects among them, jointly exerting an anti-hepatocellular carcinoma effect. Even if some herbs are changed or removed, the aforementioned excellent anti-hepatocellular carcinoma activity cannot be achieved; each component in the aforementioned traditional Chinese medicine composition is indispensable.

[0066] Experimental Example 2

[0067] The effect of the traditional Chinese medicine composition of this invention on tumor recurrence after liver cancer resection in nude mice

[0068] 1. Preparation of a mouse model of recurrence of liver carcinoma in situ after surgery

[0069] H22 cells in logarithmic growth phase were collected, washed with PBS, and the cell concentration was adjusted to 2 × 10⁻⁶. 7 0.1 mL / mouse was injected intraperitoneally into nude mice (BALB / C-nu / nu, male, 4-6 weeks old, body weight 20-24 g). After approximately 10 days, ascites fluid was collected, washed with PBS, and the cell concentration was adjusted for second-generation H22 cell inoculation. This process was repeated to collect third-generation H22 ascites cells. Before inoculation, the collected third-generation H22 ascites cells were washed three times with sterile PBS and the cell concentration was adjusted to 1×10⁻⁶ cells / mL. 70.1 mL of cell suspension was aseptically injected subcutaneously into the right axilla of BALB / c nude mice, with each mouse receiving the suspension, and the inoculation was completed within 30 minutes. Tumor size was measured daily using calipers after inoculation. Four days after tumor inoculation, nude mice with successfully implanted tumors were selected to begin PD-1 monoclonal antibody treatment. PD-1 monoclonal antibody was administered intraperitoneally at a dose of 200 μg / mouse, every two days for a total of four doses. Tumor size and general health status of the nude mice were monitored during treatment. After treatment, the mice were observed for five days, and those whose tumor volume remained unchanged or decreased during treatment but increased after drug withdrawal were selected as tumor recurrence models.

[0070] 2. Grouping and Drug Administration

[0071] Nude mice with tumor recurrence were randomly divided into a model group (Saline, n=6), a low-dose traditional Chinese medicine (TCM) group (Low, n=6), a high-dose TCM group (High, n=6), and a Western medicine control group (5-Fu, n=6). The low-dose and high-dose TCM groups were administered the decoction of the TCM composition of this invention by gavage at doses of 0.1 mL / 10 g and 0.2 mL / 10 g, respectively. The model group was administered physiological saline by gavage at a dose of 0.1 mL / 10 g, once daily. The Western medicine control group received intraperitoneal injection of 5-fluorouracil (5-Fu) at a concentration of 5 mg / mL and a dose of 40 mg / kg, once every three days. All groups received continuous treatment for 21 days.

[0072] The decoction of the composition of the present invention is prepared according to the following steps: 21 parts of Caragana korshinskii, 6 parts of red ginseng, 15 parts of Polygonum cuspidatum, 12 parts of Codonopsis pilosula, 18 parts of Lycopus lucidus, 12 parts of Semiaquilegia adoxoides, 15 parts of Viola yedoensis, 15 parts of Magnolia officinalis, and 6 parts of Glycyrrhiza uralensis (processed). Each ingredient is weighed according to the above formula, and each part by weight is 0.5g. Eight times the weight of purified water is added, and the decoction is decocted for 40 minutes. The decoction is taken, and six times the amount of purified water is added to the dregs. The decoction is decocted again for 30 minutes. Then, the two decoctions are combined, and the decoction is concentrated to one-quarter of the total volume of the decoction. The decoction is then refrigerated for later use.

[0073] 3 Observation and Detection Indicators

[0074] 3.1 Tumor growth curve

[0075] Starting from day 1 of drug administration, tumor size was measured at fixed times every 3 days, tumor volume was calculated, and a tumor growth curve was plotted. Tumor volume V = 0.5 × L (maximum tumor diameter) × S (minimum tumor diameter) 2 .

[0076] 3.2 Tumor weight

[0077] After the administration was completed, the nude mice were euthanized, the tumor tissue was removed, the surface liquid was wiped dry, and the mass of solid tumors in each group of nude mice was weighed.

[0078] 3.3 Flow cytometry detection of CD4 + CD8 + T lymphocyte level

[0079] Spleen cells were crushed, washed with physiological saline, filtered to obtain a spleen cell suspension, centrifuged, and the supernatant was discarded to obtain cell pellet. The pellet was washed with PBS and cultured at 37°C and 5% CO2 for 2 hours. Adherent cells were removed to obtain spleen lymphocytes. Blood samples were collected from the orbital cavity, anticoagulated with heparin, and stored at 4°C for blood collection. 100 μL of each tube of blood and spleen lymphocytes were added, along with 10 μL of rabbit anti-mouse CD4+CD8+ subset monoclonal antibody. The tubes were incubated at 4°C in the dark for 20 minutes, then 400 μL of PBS was added to each tube. The tubes were centrifuged at 2000 rpm for 10 minutes, the supernatant was discarded, and 0.5 mL of fixative was added. The tubes were then stored at 4°C for 30 minutes before flow cytometry analysis to detect CD4+ levels in the spleen and blood. + CD8 + T lymphocyte subset expression and CD4 + / CD8 + ratio.

[0080] 4. Statistical Analysis

[0081] The measurement data was collected using GraphPad Prism 8.0 software. This indicates that one-way ANOVA was used for comparisons among multiple groups, and P < 0.05 was considered statistically significant.

[0082] 5 Results

[0083] 5.1 Tumor growth curve

[0084] During the drug administration period, the model group mice exhibited poor mental state, reduced activity, and significantly decreased food and water intake. Compared to the model group, the mice in the low-dose and high-dose traditional Chinese medicine groups showed significantly improved mental state and significantly increased food and water intake. The mice in the western medicine control group showed no significant improvement in mental state, but slightly increased food and water intake. No mice in any group died during the experiment.

[0085] Based on the observation of tumor growth dynamics in each group of mice, it can be seen that ( Figure 2 a) In the first 6 days after administration, there was no significant difference in tumor volume among the groups of mice (P>0.05); on the 9th day after administration, the tumor volume of mice in the high-dose traditional Chinese medicine group and the western medicine control group was significantly smaller than that of the model group (P<0.05); starting from the 15th day after administration, the tumor volume of mice in the low-dose traditional Chinese medicine group, the high-dose traditional Chinese medicine group, and the western medicine control group was significantly smaller than that of the model group (P<0.05); on the 21st day after administration, the tumor volume of mice in the low-dose traditional Chinese medicine group, the high-dose traditional Chinese medicine group, and the western medicine control group was significantly lower than that of the model group (P<0.01), and the tumor weight of each administration group was significantly lower than that of the model group (P<0.01). Figure 2b).

[0086] The above results indicate that the herbal composition of this invention can significantly slow down the tumor growth rate and reduce tumor volume and weight in mice with recurrent liver carcinoma in situ after surgery, demonstrating excellent in vivo tumor suppression effects. Furthermore, compared to 5-FU, the herbal composition of this invention can better improve the survival status of mice with recurrent liver carcinoma in situ after surgery.

[0087] 5.2 CD4 in vivo + CD8 + T lymphocyte level

[0088] Compared with the model group, the CD4 counts of relapsed mice in the low-dose traditional Chinese medicine group, high-dose traditional Chinese medicine group, and Western medicine control group were significantly higher. + CD4 + / CD8 + T lymphocyte levels were significantly increased in peripheral blood and spleen (P<0.01), CD8 + The level of T lymphocytes was significantly reduced in peripheral blood and spleen (P<0.01).

[0089] CD4, as a helper T cell, acts as a central hub in the immune response to viral invasion, directing the body's fight against the virus. CD8, as a suppressor T cell, plays a role in specific immunity by recognizing and presenting antigens, and can directly inhibit and kill viruses. The expression of CD4 and CD8 T lymphocyte subsets indirectly reflects the body's immune status after surgery. These results indicate that the herbal composition of this invention can promote the expression of CD4... + T lymphocyte levels, inhibiting CD8 + By regulating T lymphocyte levels, the immune function can be improved, immune imbalance in relapsed mice can be improved, and immune function can be strengthened after liver cancer surgery, thereby achieving the goal of preventing liver cancer recurrence.

[0090] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A traditional Chinese medicine composition for preventing and treating postoperative recurrence of liver cancer, characterized in that, The traditional Chinese medicine composition is made from the following traditional Chinese medicinal materials in parts by weight: 10-30 parts of Caragana korshinskii, 1-15 parts of red ginseng, 5-20 parts of Polygonum cuspidatum, 5-20 parts of Codonopsis pilosula, 5-20 parts of Lycopus lucidus, 5-20 parts of Semiaquilegia adoxoides, 5-20 parts of Viola yedoensis, 5-20 parts of Magnolia officinalis, and 1-15 parts of prepared licorice root.

2. The traditional Chinese medicine composition for preventing postoperative recurrence of liver cancer as described in claim 1, characterized in that, The traditional Chinese medicine composition is made from the following traditional Chinese medicinal materials in parts by weight: 15-25 parts of Caragana korshinskii, 3-10 parts of red ginseng, 10-20 parts of Polygonum cuspidatum, 10-20 parts of Codonopsis pilosula, 10-20 parts of Lycopus lucidus, 10-20 parts of Semiaquilegia adoxoides, 10-20 parts of Viola yedoensis, 10-20 parts of Magnolia officinalis, and 1-10 parts of prepared licorice root.

3. The traditional Chinese medicine composition for preventing postoperative recurrence of liver cancer as described in claim 1, characterized in that, The traditional Chinese medicine composition is made from the following traditional Chinese medicinal materials in parts by weight: 21 parts of Caragana korshinskii, 6 parts of red ginseng, 15 parts of Polygonum cuspidatum, 12 parts of Codonopsis pilosula, 18 parts of Lycopus lucidus, 12 parts of Semiaquilegia adoxoides, 15 parts of Viola yedoensis, 15 parts of Magnolia officinalis, and 6 parts of Glycyrrhiza uralensis (processed).

4. The use of the traditional Chinese medicine composition according to any one of claims 1 to 3 in the preparation of a drug for preventing postoperative recurrence of liver cancer, characterized in that, The drug is made from the Chinese medicinal materials described in any one of claims 1 to 3 by weight, prepared according to conventional methods, and supplemented with pharmaceutically acceptable excipients, into any one of the following oral dosage forms: granules, formulated granules, oral liquids, capsules, tablets, pills, and decoctions.

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