Application of traditional Chinese medicine composition in preparation of medicine for resisting colon cancer

Abstract

The invention relates to a novel application of hemorrhoid eliminating medicine in resisting colon cancer. The medicinal materials in the hemorrhoid eliminating medicine formula for internal and external hemorrhoids are found to have a remarkable synergistic anti-colon cancer effect for the first time, and experiments find that the hemorrhoid eliminating medicine formula can remarkably improve the weight and colon length of a colon cancer model mouse, reduce the number of tumors in colon tissues, improve the pathological state of the colon tissues and improve the curative effect of the colon cancer. The levels of colon tissue inflammatory factors and serum tumor markers are reduced, and the traditional Chinese medicine composition has remarkable relieving and treating effects on colon cancer. Moreover, compared with the existing anti-colon cancer drugs, the hemorrhoid eliminating drug has the advantages of low toxicity and safety, reduces the toxic and side effects of colon cancer drugs, provides a brand new drug choice for the treatment of colon cancer, and has important scientific value and clinical application prospect.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceuticals, relates to a traditional Chinese medicine composition, and specifically relates to the application of a traditional Chinese medicine composition in the preparation of a drug for anti-colorectal cancer. Background Art

[0002] Colorectal cancer is one of the malignant tumors with high incidence and mortality rates globally. Currently, clinical treatment mainly focuses on surgical resection combined with chemotherapy, targeted therapy, or immunotherapy, but there are the following limitations: Chemotherapy drugs (such as 5-fluorouracil, oxaliplatin) are prone to cause systemic side effects such as myelosuppression and gastrointestinal reactions, and the problem of drug resistance is common; Although targeted drugs (such as anti-EGFR antibodies, anti-angiogenic drugs) can improve the therapeutic effect, the applicable population is limited (depending on specific gene mutations), and the treatment cost is high; Immune checkpoint inhibitors are effective for patients with microsatellite highly unstable (MSI-H), but only account for about 15% of colorectal cancer cases, and there is a risk of immune-related adverse reactions. Therefore, there is an urgent need to develop new and low-toxic anti-tumor drugs.

[0003] Xiaozhiling is a compound traditional Chinese medicine composed of Galla Chinensis, Ampelopsis japonica, Selaginella tamariscina, Sanguisorba officinalis, Sophora japonica, and Cholic acid. The Xiaozhiling formula has the functions of astringing and stopping bleeding, and detoxifying and astringing sores, can relieve the pain caused by prolapse, incarceration, swelling, and erosion necrosis of hemorrhoids, and is commonly used in internal and external hemorrhoids clinically. There is no relevant research publicly reporting its application in the anti-tumor field. Summary of the Invention

[0004] Based on this, the purpose of the present invention is to provide a new drug for anti-colorectal cancer.

[0005] The technical solutions to achieve the above purpose are as follows.

[0006] The present invention provides the application of a traditional Chinese medicine composition in the preparation of a drug for anti-colorectal cancer, and the active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 120 - 380 parts of Galla Chinensis, 700 - 1300 parts of Sophora japonica, 700 - 1300 parts of Ampelopsis japonica, 700 - 1300 parts of Selaginella tamariscina, 700 - 1300 parts of Sanguisorba officinalis, and 12 - 38 parts of Cholic acid.

[0007] The present invention has the following beneficial effects:

[0008] The present invention for the first time discovers that a traditional Chinese medicine composition composed of Chinese gall, Ampelopsis japonica, Selaginella tamariscina, Sanguisorba officinalis, Sophora japonica, and cholic acid in a specific ratio (i.e., the Xiaozhiling drug for internal and external hemorrhoids) has a significant alleviating and therapeutic effect on colon cancer. Experiments find that it can significantly increase the body weight and colon length of colon cancer model mice, reduce the number of tumors in the colon tissue, improve the pathological state of the colon tissue, and lower the levels of inflammatory factors in the colon tissue and serum tumor markers; moreover, its anti-colon cancer effect is significantly superior to that of the Sanguisorba officinalis - Sophora japonica herb pair, the Chinese gall - Selaginella tamariscina herb pair, and single herbs such as Chinese gall, Sophora japonica, Selaginella tamariscina, Ampelopsis japonica, and Sanguisorba officinalis; each of the medicinal materials therein has a significant synergistic anti-colon cancer effect.

[0009] Xiaozhiling is a compound preparation of traditional Chinese medicine widely used clinically, and its safety has been widely clinically verified. Compared with existing anti-colon cancer drugs, it has the advantage of low toxicity and safety, reducing the toxic and side effects of colon cancer medications, providing a new drug option for the treatment of colon cancer, and having important scientific value and clinical application prospects. Brief Description of the Drawings

[0010] Figure 1 For the pathological observation results and scores of the colon tissues of each group of mice; among them, A is the normal group, B is the model group, C is the Xiaozhiling formula 1 group, D is the Xiaozhiling formula 2 group, E is the Xiaozhiling formula 3 group, F is the Xiaozhiling formula 4 group, G is the Xiaozhiling formula 5 group, H is the inflammatory score of the colon tissues of each group; compared with the normal group, a P < 0.05; compared with the model group, b P < 0.05; compared with the Xiaozhiling formula 1 group, c P < 0.05.

[0011] Figure 2 For the pathological observation results and scores of the colon tissues of each group of mice; among them, A is the normal group, B is the model group, C is the Xiaozhiling 1 group, D is the Sanguisorba officinalis - Sophora japonica group, E is the Chinese gall - Selaginella tamariscina group, F is the Ampelopsis japonica - cholic acid group, G is the Chinese gall group, H is the Sophora japonica group, I is the Selaginella tamariscina group, L is the Sanguisorba officinalis group, M is the Ampelopsis japonica group, N is the cholic acid group, O is the inflammatory score of the colon tissues of each group; compared with the normal group, a P < 0.05; compared with the model group, b P < 0.05; compared with the Xiaozhiling formula 1 group, c P < 0.05. Detailed Embodiments

[0012] To facilitate the understanding of the present invention, the present invention will be described more comprehensively below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, these embodiments are provided to make the understanding of the disclosed content of the present invention more thorough and comprehensive.

[0013] In the following examples, the experimental methods without specific conditions are generally carried out under conventional conditions or according to the conditions recommended by the manufacturers. All common chemical reagents used in the examples are commercially available products.

[0014] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those skilled in the technical field to which the present invention pertains. The terms used in the description of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used in the present invention includes any and all combinations of one or more of the related listed items.

[0015] In addition, as used in the present invention, the term "or" is the inclusive "or" symbol and is equivalent to the term "and / or", unless the context clearly dictates otherwise. The term "based on" is not exclusive and allows for other factors not described, unless the context clearly dictates otherwise. Further, throughout the specification, the meanings of "a", "an", and "the" include plural referents. The meaning of "in..." includes "in..." and "on...".

[0016] The present invention provides an application of a traditional Chinese medicine composition in the preparation of a drug for treating colon cancer. The active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 120 - 380 parts of Chinese gall, 700 - 1300 parts of Japanese pagoda tree flower, 700 - 1300 parts of Ampelopsis japonica, 700 - 1300 parts of Selaginella tamariscina, 700 - 1300 parts of Sanguisorba officinalis, and 12 - 38 parts of cholic acid.

[0017] In some of the embodiments, the active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 150 - 350 parts of Chinese gall, 800 - 1200 parts of Japanese pagoda tree flower, 800 - 1200 parts of Ampelopsis japonica, 800 - 1200 parts of Selaginella tamariscina, 800 - 1200 parts of Sanguisorba officinalis, and 15 - 35 parts of cholic acid.

[0018] In some of the embodiments, the mass ratio of the Japanese pagoda tree flower, Ampelopsis japonica, Selaginella tamariscina, and Sanguisorba officinalis is 1:0.9 - 1.1:0.9 - 1.1:0.9 - 1.1.

[0019] In some of the embodiments, the mass ratio of the Japanese pagoda tree flower, Ampelopsis japonica, Selaginella tamariscina, and Sanguisorba officinalis is 1:1:1:1.

[0020] In some of the embodiments, the mass ratio of the Chinese gall to the Japanese pagoda tree flower is 1:3 - 6.

[0021] In some of the embodiments, the mass ratio of the Chinese gall to the Japanese pagoda tree flower is 1:3.4 - 5.4, such as 1:4, 15:80, 35:120.

[0022] In some of these embodiments, the mass ratio of Chinese gallnut to cholic acid is 1:0.08 - 0.12.

[0023] In some of these embodiments, the mass ratio of Chinese gallnut to cholic acid is 1:0.1.

[0024] For example, the active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 250 parts of Chinese gallnut, 1000 parts of sophora flower, 1000 parts of ampelopsis root, 1000 parts of selaginella, 1000 parts of sanguisorba root, and 25 parts of cholic acid;

[0025] Or, the active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 150 parts of Chinese gallnut, 800 parts of sophora flower, 800 parts of ampelopsis root, 800 parts of selaginella, 800 parts of sanguisorba root, and 15 parts of cholic acid;

[0026] Or, the active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 350 parts of Chinese gallnut, 1200 parts of sophora flower, 1200 parts of ampelopsis root, 1200 parts of selaginella, 1200 parts of sanguisorba root, and 35 parts of cholic acid.

[0027] In some of these embodiments, the active ingredients of the traditional Chinese medicine composition are prepared by the following method: Take the Chinese gallnut, sophora flower, ampelopsis root, selaginella, and sanguisorba root, mix them, soak them in water, then decoct them, combine the decocted liquid, and then add the cholic acid and mix well to obtain.

[0028] In some of these embodiments, decoct 2 - 4 times. The water addition amount for the first decoction is 6 - 8 times the total weight of the medicinal materials, and the water addition amount for each subsequent decoction is 4 - 6 times the total weight of the medicinal materials.

[0029] In some of these embodiments, decoct 3 times. The water addition amount for the first decoction is 7 times the total weight of the medicinal materials, and the water addition amount for each subsequent decoction is 5 times the total weight of the medicinal materials.

[0030] In some of these embodiments, the decoction time for each time is 20 min - 40 min.

[0031] In some of these embodiments, the decoction time for each time is 25 min - 35 min.

[0032] In some of these embodiments, the soaking time is 20 min - 40 min.

[0033] In some of these embodiments, the soaking time is 25 min - 35 min.

[0034] In some of these embodiments, the traditional Chinese medicine composition further includes pharmaceutically acceptable excipients, such as sodium carboxymethylcellulose, etc.

[0035] In some of these embodiments, the dosage form of the anti-colorectal cancer drug is tablets, injections, capsules, granules, powders, mixtures, emulsions, decoctions or suspensions.

[0036] The following further elaborates on the present invention in conjunction with specific embodiments.

[0037] Example 1 Effects of Compound Xiaozhiling with Different Proportions on Mice with Colorectal Cancer Models

[0038] 1. Method

[0039] 1.1 Animal Grouping and Model Construction

[0040] SPF-grade male C57BL / 6J mice, weighing 20 ± 2 g. One week before the experiment, they were adaptively raised under the conditions of free access to food and water, a temperature of 20 ± 2 °C, a humidity of 45% - 55%, and a 12-hour light cycle. They were randomly divided into a normal group, a model group, and Compound Xiaozhiling groups 1, 2, 3, 4, and 5 (600 mg crude drug / kg), with 10 mice in each group. On the first day, except for the normal group, the other groups were intraperitoneally injected with 10 mg / kg azoxymethane (AOM), and immediately after the injection, the drinking water was replaced with an aqueous solution containing 2% dextran sulfate sodium (DSS) for 1 week, and then the DSS aqueous solution was replaced with pure water for 2 weeks. This was one cycle, and such a cycle was repeated 3 times in total. The mice in the normal group were intraperitoneally injected with normal saline and drank water freely.

[0041] 1.2 Drug Preparation and Administration

[0042] The compositions of each medicinal material in Compound Xiaozhiling 1, 2, 3, 4, and 5 are shown in Table 1, and the preparation method is as follows: The medicinal materials of Galla Chinensis, Ampelopsis japonica, Selaginella tamariscina, Sanguisorba officinalis, and Sophora japonica flower were soaked in water 7 times the total weight of the medicinal materials for 30 minutes and then decocted for 30 minutes; after filtration, the medicinal residues were decocted twice more, with the water addition amount each time being 5 times the total weight of the medicinal materials, and each decoction was for 30 minutes; the decoction liquids after the three decoctions were combined, and then the corresponding weight parts of cholic acid and sodium carboxymethylcellulose (the final concentration in the decoction liquid was 0.5 wt%) were added, dissolved and mixed evenly for standby.

[0043] From the 4th week, the mice in the administration groups were gavaged once a day, and the model group and the normal group were gavaged with pure water. In the 11th week, the mice in each group were bled from the eyeballs and then sacrificed by cervical dislocation, and biological samples were collected.

[0044] Table 1 Different Proportion Compositions of Compound Xiaozhiling (parts by weight)

[0045]

[0046]

[0047] 1.3 General State Observation

[0048] During the establishment of the model and administration of drugs, observe the mental state, hair, diet and water intake of the mice, and record the daily body weight of the mice. After sacrificing the mice, measure the colon length of the mice.

[0049] 1.4 Pathological detection and scoring of mouse colon

[0050] After the experiment, sacrifice the mice, collect their colon tissues. After embedding and paraffin section processing, observe the pathological changes of the colon tissues by hematoxylin-eosin (HE) staining method, including inflammation, ulcer, polyp and tumor formation. The pathological morphology of mouse colon tissues was scored for inflammation according to the Dieleman standard: 0 point, no inflammation; 1 point, basal crypt damage > 1 / 3; 2 point, basal crypt damage > 2 / 3; 3 point, complete loss of crypts, accompanied by a small amount of inflammatory cell infiltration; 4 point, mucosal erosion, ulcer, accompanied by a large amount of inflammatory cell infiltration.

[0051] 1.5 Determination of inflammatory factors in mouse colon tissues

[0052] Refer to the kit instructions and use ELISA method to detect the expression levels of IL-1β, TNF-α and IL-6 in colon tissues.

[0053] 1.6 Levels of serum tumor markers CA199, CEA and CA724 in mice

[0054] After the experiment, let the blood of the mice in the normal group and the model group collected stand at room temperature for 1 h, centrifuge at 3000 rpm / min at 4 °C for 15 min, take the supernatant, and refer to the kit instructions to use ELISA method to detect the levels of tumor markers CA199, CEA and CA724 in the serum of mice.

[0055] 2. Results

[0056] 2.1 Body weight and pathological observation of colon tissues in each group of mice

[0057] The detection results of the body weight and colon conditions of each group of mice (see Table 2) showed that: compared with the normal group of mice, the body weight of the model group of mice decreased significantly (P < 0.05), the colon length was significantly shortened (P < 0.05), a certain number of tumor-like protrusions of different sizes could be seen on the mucosal surface (P < 0.05), most mice had bloody stools, and severe cases were accompanied by anal prolapse symptoms; compared with the model group, the body weights of the mice in the Xiaozhiling Prescription 1, 2, and 3 groups increased significantly (P < 0.05), the colon length increased significantly (P < 0.05), and the number of colon tumors decreased significantly (P < 0.05), and only a small number of mice still had bloody stools; compared with the Xiaozhiling Prescription 1 group, there were no statistically significant differences in the body weight, colon length, and number of colon tumors of the mice in the Xiaozhiling Prescription 2 and 3 groups (P > 0.05), and the body weights of the Xiaozhiling Prescription 4 and 5 groups decreased significantly (P < 0.05), the colon length was significantly shortened (P < 0.05), and the number of mucosal colon tumors increased significantly (P < 0.05).

[0058] The HE staining results of the colon tissues of each group of mice (see Figure 1 ) showed that the colon mucosal epithelium of the normal group of mice was intact, the glandular structure was arranged regularly, the crypts were normal, and there was no congestion, edema, erosion, ulcer, or inflammatory cell infiltration; the colon mucosa of the model group of mice was inflamed, congested, and edematous, the glandular structure was loose, there was inflammatory cell infiltration, the cells were of different sizes and shapes, arranged disorderly, the polarity was lost, and there were manifestations of atypical hyperplasia to varying degrees such as large and deeply stained nuclei, increased nuclear-cytoplasmic ratio, irregular nuclear shape, and increased mitotic figures. Compared with the normal group, the inflammation score of the colon tissue was significantly increased (P < 0.05); compared with the model group, the disorder of the glandular structure in the mice of the Xiaozhiling Prescription 1, 2, and 3 groups was reduced, the inflammatory infiltration was reduced, the cell size, shape, and arrangement were more regular, and the inflammation score of the colon tissue was significantly reduced (P < 0.05); compared with the Xiaozhiling Prescription 1 group, there were no statistically significant differences in the inflammation scores of the colon tissues of the Xiaozhiling Prescription 2 and 3 groups (P > 0.05), and the inflammation scores of the colon tissues of the mice in the Xiaozhiling Prescription 4 and 5 groups were significantly increased (P < 0.05).

[0059] Table 2 Body weight, colon length, and number of colon tumors of each group of mice (x±s, n = 10)

[0060]

[0061]

[0062] Note: Compared with the normal group, a P < 0.05; compared with the model group, b P < 0.05; compared with the Xiaozhiling Prescription 1 group, c P < 0.05

[0063] 2.2 Expression levels of inflammatory factors in the colon tissues of each group of mice

[0064] The results of measuring the expression levels of inflammatory factors in the colon tissues of mice in each group (see Table 3) showed that compared with the normal group, the expression levels of IL-1β, IL-6, and TNF-α in the colon tissues of mice in the model group were significantly increased (P < 0.05); compared with the model group, the expression levels of IL-1β, IL-6, and TNF-α in the colon tissues of mice in the Xiaozhiling Prescription 1, 2, 3, 4, and 5 groups were significantly decreased (P < 0.05); compared with the Xiaozhiling Prescription 1 group, there was no significant difference in the expression levels of IL-1β, IL-6, and TNF-α in the colon tissues of mice in the Xiaozhiling Prescription 2 and 3 groups (P > 0.05), and the expression levels of IL-1β, IL-6, and TNF-α in the colon tissues of mice in the Xiaozhiling Prescription 4 and 5 groups were significantly increased (P < 0.05).

[0065] Table 3 Levels of inflammatory factors in the colon tissues of mice in each group ( n = 10, pg / mg)

[0066]

[0067] Note: Compared with the normal group, a P < 0.05; compared with the model group, b P < 0.05; compared with the Xiaozhiling Prescription 1 group, c P < 0.05

[0068] 2.3 Levels of serum tumor markers in mice in each group

[0069] The results of measuring the levels of serum tumor markers in mice in each group (see Table 4) showed that compared with the normal group, the levels of serum tumor markers CA199, CEA, and CA724 in the model group were significantly increased (P < 0.05), which indicated the occurrence and formation of tumors in the model mice. Compared with the model group, the levels of serum tumor markers CA199, CEA, and CA724 in the Xiaozhiling Prescription 1, 2, 3, 4, and 5 groups were significantly decreased (P < 0.05); compared with the Xiaozhiling Prescription 1 group, there was no significant difference in the levels of serum tumor markers CA199, CEA, and CA724 in the Xiaozhiling Prescription 2 and 3 groups (P > 0.05), and the levels of serum tumor markers CA199, CEA, and CA724 in the Xiaozhiling Prescription 4 and 5 groups were significantly increased (P < 0.05).

[0070] Table 4 Levels of serum tumor markers in mice in each group ( n = 10)

[0071]

[0072] Note: Compared with the normal group, a P < 0.05; compared with the model group, b P < 0.05; compared with the Xiaozhiling Prescription 1 group, cP < 0.05

[0073] 3. Conclusion

[0074] The traditional Chinese medicine composition (i.e., Xiaozhiling drug for internal and external hemorrhoids) composed of Chinese gallnut, Ampelopsis japonica, Selaginella tamariscina, Sanguisorba officinalis, Sophora japonica, and cholic acid in a specific ratio has a significant anti - colon cancer effect. It can significantly increase the body weight and colon length of colon cancer model mice, reduce the number of tumors in the colon tissue, improve the pathological state of the colon tissue, and reduce the levels of inflammatory factors in the colon tissue and serum tumor markers.

[0075] Effect of the disassembled formula of Xiaozhiling on colon cancer model mice in the control group

[0076] 1. Method

[0077] 1.1 Animal grouping and model construction

[0078] SPF - level male C57BL / 6J mice, weighing 20 ± 2 g. One week before the experiment, they were adaptively fed under the conditions of free access to food and water, temperature 20 ± 2 °C, humidity 45% - 55%, and a 12 - h light cycle. They were randomly divided into a normal group, a model group, a Xiaozhiling formula 1 group (600 mg crude drug / kg), a Sanguisorba officinalis - Sophora japonica group (600 mg crude drug / kg), a Chinese gallnut - Selaginella tamariscina group (600 mg crude drug / kg), an Ampelopsis japonica - cholic acid group (600 mg crude drug / kg), a Chinese gallnut group (600 mg crude drug / kg), a Sophora japonica group (600 mg crude drug / kg), an Ampelopsis japonica group (600 mg crude drug / kg), a Selaginella tamariscina group (600 mg crude drug / kg), a Sanguisorba officinalis group (600 mg crude drug / kg), and a cholic acid group (600 mg cholic acid / kg), with 10 mice in each group. On the first day, except for the normal group, the other groups were intraperitoneally injected with 10 mg / kg azoxymethane (AOM), and immediately after injection, the drinking water was replaced with an aqueous solution containing 2% dextran sulfate sodium (DSS) for 1 week, and then the DSS aqueous solution was replaced with pure water for 2 weeks. This was one cycle, and such a cycle was repeated 3 times in total. The mice in the normal group were intraperitoneally injected with normal saline and drank water freely.

[0079] 1.2 Drug preparation and administration method

[0080] The preparation of Xiaozhiling formula 1 was the same as that in "Example 1".

[0081] The preparation method of the medicinal liquid in the Sanguisorba officinalis - Sophora japonica group was as follows: Take 1000 parts by weight of Sophora japonica and 1000 parts by weight of Sanguisorba officinalis and mix them; soak the medicinal materials in water 7 times the total weight of the medicinal materials for 30 min and then decoct for 30 min; filter, and decoct the medicinal residues twice more, with the water addition amount each time being 5 times the total weight of the medicinal materials, and decoct for 30 min each time; combine the medicinal liquids after the three decoctions and set aside.

[0082] The preparation method of the liquid medicine of the Galla Chinensis-Selaginella tamariscina group is as follows: Take 250 parts by weight of Galla Chinensis and 1000 parts by weight of Selaginella tamariscina and mix them; soak the medicinal materials in water 7 times the total weight of the medicinal materials for 30 min and then decoct for 30 min; filter, take the medicinal residues and decoct them twice more, with the water addition amount each time being 5 times the total weight of the medicinal materials, and decoct for 30 min each time; combine the liquid medicines after the three decoctions and reserve for use.

[0083] The preparation method of the liquid medicine of the Ampelopsis japonica-Cholic acid group is as follows: Take 1000 parts by weight of Ampelopsis japonica, soak the medicinal materials in water 7 times the total weight of the medicinal materials for 30 min and then decoct for 30 min; filter, take the medicinal residues and decoct them twice more, with the water addition amount each time being 5 times the total weight of the medicinal materials, and decoct for 30 min each time; combine the liquid medicines after the three decoctions, then add 25 parts by weight of cholic acid and sodium carboxymethylcellulose (the final concentration in the liquid medicine is 0.5 wt%), dissolve and mix evenly, and reserve for use.

[0084] The preparation method of the liquid medicine of the Galla Chinensis, Sophora japonica, Ampelopsis japonica, Selaginella tamariscina, Sanguisorba officinalis group is as follows: Soak the medicinal materials in water 7 times the total weight of the medicinal materials for 30 min and then decoct for 30 min; filter, take the medicinal residues and decoct them twice more, with the water addition amount each time being 5 times the total weight of the medicinal materials, and decoct for 30 min each time; combine the liquid medicines after the three decoctions and reserve for use.

[0085] Preparation of the cholic acid group liquid medicine: Take cholic acid and dissolve it in water containing 0.5 wt% sodium carboxymethylcellulose and reserve for use.

[0086] From the 4th week, the mice in the administration group were given medicine by gavage once a day, and the mice in the model group and the normal group were given pure water by gavage. In the 11th week, the mice in each group were bled from the eyeballs, and then sacrificed by cervical dislocation, and the biological samples were collected.

[0087] 1.3 General state observation

[0088] During the modeling and administration period, observe the mental state, hair, diet and drinking water of the mice, and record the daily body weight of the mice. After sacrificing the mice, measure the colon length of the mice.

[0089] 1.4 Pathological detection and scoring of the mouse colon

[0090] After the experiment, the mice were sacrificed, and their colon tissues were collected. After embedding and paraffin section processing, observe the pathological changes of the colon tissues by hematoxylin-eosin (HE) staining method, including inflammation, ulcer, polyp and tumor formation. The pathological morphology of the mouse colon tissues was scored for inflammation according to the Dieleman standard: 0 point, no inflammation; 1 point, basal crypt damage > 1 / 3; 2 point, basal crypt damage > 2 / 3; 3 point, complete crypt defect, accompanied by a small amount of inflammatory cell infiltration; 4 point, mucosal erosion, ulcer, accompanied by a large amount of inflammatory cell infiltration.

[0091] 1.5 Determination of inflammatory factors in mouse colon tissues

[0092] The expression levels of IL-1β, TNF-α, and IL-6 in colon tissues were detected by ELISA according to the kit instructions.

[0093] 1.6 Levels of serum tumor markers CA199, CEA, and CA724 in mice

[0094] After the experiment, the blood samples collected from the normal group and the model group mice were allowed to stand at room temperature for 1 h, centrifuged at 3000 rpm / min at 4 °C for 15 min, and the supernatant was taken. The levels of tumor markers CA199, CEA, and CA724 in the mouse serum were detected by ELISA according to the kit instructions.

[0095] 2. Results

[0096] 2.1 Body weight and pathological observation of colon tissues in each group of mice

[0097] The detection results of the body weight and colon conditions of each group of mice (see Table 5) showed that: compared with the normal group mice, the body weight of the model group mice decreased significantly (P < 0.05), the colon length was significantly shortened (P < 0.05), a certain number of tumor-like protrusions of different sizes could be seen on the mucosal surface (P < 0.05), most mice had bloody stools, and severe cases were accompanied by rectal prolapse symptoms; compared with the model group, the body weight of the Xiaozhiling Prescription 1 group, Sanguisorba-Floral Sophora group, Galla Chinensis-Selaginella group, Galla Chinensis group, Floral Sophora group, Selaginella group, and Sanguisorba group mice increased significantly (P < 0.05), the colon length increased significantly (P < 0.05), and the number of colon tumors decreased significantly (P < 0.05). Only a small number of mice still had bloody stools, but there were no significant differences in the body weight, colon length, and number of colon tumors among the Ampelopsis-Glycholic Acid group, Ampelopsis group, and Glycholic Acid group mice (P > 0.05); compared with the Xiaozhiling Prescription 1 group, the body weight of the Sanguisorba-Floral Sophora group, Galla Chinensis-Selaginella group, Ampelopsis-Glycholic Acid group, Galla Chinensis group, Floral Sophora group, Selaginella group, Sanguisorba group, Ampelopsis group, and Glycholic Acid group mice decreased significantly (P < 0.05), the colon length was significantly shortened (P < 0.05), and the number of mucosal colon tumors increased significantly (P < 0.05).

[0098] The HE staining results of the colon tissues of each group of mice (see Figure 2)It was shown that the colonic mucosa epithelium of mice in the normal group was intact, the glandular structure was arranged regularly, the crypts were normal, and there was no congestion, edema, erosion, ulcer or inflammatory cell infiltration; the colonic mucosa of mice in the model group showed inflammatory congestion and edema, the glandular structure was loose, there was inflammatory cell infiltration, the cells were of different sizes and shapes, arranged disorderly, the polarity was lost, and there were manifestations of atypical hyperplasia to varying degrees such as large and deeply stained nuclei, increased nuclear-cytoplasmic ratio, irregular nuclear shape, and increased mitotic figures. Compared with the normal group, the inflammatory score of the colonic tissue was significantly increased (P < 0.05); compared with the model group, the glandular structure disorder of mice in the Xiaozhiling group, the Sanguisorba-Floral Sophora group, the Galla Chinensis-Selaginella group, the Galla Chinensis group, the Floral Sophora group, the Selaginella group, and the Sanguisorba group was alleviated, the inflammatory infiltration was reduced, the cell size, shape and arrangement were more regular, and the inflammatory score of the colonic tissue was significantly decreased (P < 0.05), but there were no obvious pathological changes in the colonic tissue of mice in the Ampelopsis-Glycholic acid group, the Ampelopsis group, and the Glycholic acid group, and there was no significant difference in the inflammatory score (P > 0.05); compared with the Xiaozhiling formula 1 group, the inflammatory scores of the colonic tissue of mice in the Sanguisorba-Floral Sophora group, the Galla Chinensis-Selaginella group, the Ampelopsis-Glycholic acid group, the Galla Chinensis group, the Floral Sophora group, the Selaginella group, the Sanguisorba group, the Ampelopsis group, and the Glycholic acid group were significantly increased (P < 0.05).

[0099] Table 5 Body weight, colonic length and number of colonic tumors in each group of mice( n = 10)

[0100]

[0101]

[0102] Note: Compared with the normal group, a P < 0.05; compared with the model group, b P < 0.05; compared with the Xiaozhiling formula 1 group, c P < 0.05

[0103] 2.2 Expression levels of inflammatory factors in colonic tissues of mice in each group

[0104] The determination results of the expression levels of inflammatory factors in the colon tissues of mice in each group (see Table 6) showed that compared with the normal group, the expression levels of IL-1β, IL-6, and TNF-α in the colon tissues of mice in the model group were significantly increased (P < 0.05); compared with the model group, the expression levels of IL-1β, IL-6, and TNF-α in the colon tissues of mice in the Xiaozhiling Recipe 1 group, Sanguisorba-Floral Sophora group, Galla Chinensis-Selaginella group, Galla Chinensis group, Floral Sophora group, Selaginella group, and Sanguisorba group were significantly decreased (P < 0.05), but there were no significant differences in the expression levels of IL-1β, IL-6, and TNF-α in the colon tissues of mice in the Ampelopsis-Glycodeoxycholic acid group, Ampelopsis group, and Glycodeoxycholic acid group (P > 0.05); compared with the Xiaozhiling Recipe 1 group, the expression levels of IL-1β, IL-6, and TNF-α in the colon tissues of mice in the Sanguisorba-Floral Sophora group, Galla Chinensis-Selaginella group, Ampelopsis-Glycodeoxycholic acid group, Galla Chinensis group, Floral Sophora group, Selaginella group, Sanguisorba group, Ampelopsis group, and Glycodeoxycholic acid group were significantly increased (P

[0105] < 0.05).

[0106] Table 6 Levels of inflammatory factors in the colon tissues of mice in each group ( n = 10, pg / mg)

[0107]

[0108]

[0109] Note: Compared with the normal group, a P < 0.05; compared with the model group, b P < 0.05; compared with the Xiaozhiling Recipe 1 group, c P < 0.05

[0110] 2.3 Serum tumor marker levels of mice in each group

[0111] The determination results of serum tumor markers in each group of mice (see Table 7) showed that, compared with the normal group, the levels of serum tumor markers CA199, CEA, and CA724 in the model group of mice were significantly increased (P < 0.05), which suggested the occurrence and formation of tumors in the model mice. Compared with the model group, the levels of serum tumor markers CA199, CEA, and CA724 in the Xiaozhiling Prescription 1 group, Sanguisorba-flos Sophorae group, Galla Chinensis-Selaginella tamariscina group, Galla Chinensis group, flos Sophorae group, Selaginella tamariscina group, and Sanguisorba group of mice were significantly decreased (P < 0.05), but there were no significant differences in the levels of serum tumor markers CA199, CEA, and CA724 in the Ampelopsis chinensis-cholic acid group, Ampelopsis chinensis group, and cholic acid group of mice (P > 0.05); compared with the Xiaozhiling Prescription 1 group, the levels of serum tumor markers CA199, CEA, and CA724 in the Sanguisorba-flos Sophorae group, Galla Chinensis-Selaginella tamariscina group, Ampelopsis chinensis-cholic acid group, Galla Chinensis group, flos Sophorae group, Selaginella tamariscina group, Sanguisorba group, Ampelopsis chinensis group, and cholic acid group of mice were significantly increased (P < 0.05).

[0112] Table 7 Serum tumor marker levels of each group of mice ( n = 10)

[0113]

[0114]

[0115] Note: Compared with the normal group, a P < 0.05; compared with the model group, b P < 0.05; compared with the Xiaozhiling Prescription 1 group, c P < 0.05

[0116] 3. Conclusions

[0117] The above results indicated that Galla Chinensis, flos Sophorae, Selaginella tamariscina, and Sanguisorba in the Xiaozhiling Prescription had certain anti-colorectal cancer effects, while Ampelopsis chinensis and cholic acid had no anti-colorectal cancer effects; however, when Ampelopsis chinensis, cholic acid were combined with Galla Chinensis, flos Sophorae, Selaginella tamariscina, and Sanguisorba, the anti-colorectal cancer effect of the compound prescription could be significantly improved, and the anti-colorectal cancer effect of the whole Xiaozhiling Prescription was significantly better than that of each single herb used alone and the combination of each pair of herbs. It showed that the components in the traditional Chinese medicine composition composed of Galla Chinensis, flos Sophorae, Selaginella tamariscina, Sanguisorba, Ampelopsis chinensis, and cholic acid of the present invention were rationally combined, and had a significant synergistic effect on the treatment of colorectal cancer.

[0118] The above-described embodiments merely represent several implementation manners of the present invention. The description thereof is relatively specific and detailed, but it should not be construed as a limitation on the scope of the invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all fall within the protection scope of the present invention. Therefore, the protection scope of the present invention patent shall be subject to the appended claims.

Claims

1. Use of a traditional Chinese medicine composition in the preparation of a drug for anti - colon cancer, characterized in that, The active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 120-380 parts of Chinese gall, 700-1300 parts of Japanese pagoda tree flower, 700-1300 parts of Ampelopsis japonica, 700-1300 parts of Selaginella tamariscina, 700-1300 parts of Sanguisorba officinalis, and 12-38 parts of cholic acid.

2. The application according to claim 1, characterized in that The active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 150-350 parts of Chinese gall, 800-1200 parts of Japanese pagoda tree flower, 800-1200 parts of Ampelopsis japonica, 800-1200 parts of Selaginella tamariscina, 800-1200 parts of Sanguisorba officinalis, and 15-35 parts of cholic acid.

3. The application according to claim 1, characterized in that, The mass ratio of the Japanese pagoda tree flower, Ampelopsis japonica, Selaginella tamariscina and Sanguisorba officinalis is 1: 0.9-1.1: 0.9-1.1: 0.9-1.1; and / or, the mass ratio of the Chinese gall to the Japanese pagoda tree flower is 1: 3-6; and / or, the mass ratio of the Chinese gall to the cholic acid is 1: 0.08-0.

12.

4. The application according to claim 3, wherein The mass ratio of the Japanese pagoda tree flower, Ampelopsis japonica, Selaginella tamariscina and Sanguisorba officinalis is 1: 1: 1: 1; and / or, the mass ratio of the Chinese gall to the Japanese pagoda tree flower is 1: 3.4-5.4; and / or, the mass ratio of the Chinese gall to the cholic acid is 1: 0.

1.

5. The application according to claim 1, characterized in that, The active ingredients of the traditional Chinese medicine composition are prepared from the following raw materials in parts by weight: 250 parts of Chinese gall, 1000 parts of Japanese pagoda tree flower, 1000 parts of Ampelopsis japonica, 1000 parts of Selaginella tamariscina, 1000 parts of Sanguisorba officinalis, and 25 parts of cholic acid; or, 150 parts of Chinese gall, 800 parts of Japanese pagoda tree flower, 800 parts of Ampelopsis japonica, 800 parts of Selaginella tamariscina, 800 parts of Sanguisorba officinalis, and 15 parts of cholic acid; or, 350 parts of Chinese gall, 1200 parts of Japanese pagoda tree flower, 1200 parts of Ampelopsis japonica, 1200 parts of Selaginella tamariscina, 1200 parts of Sanguisorba officinalis, and 35 parts of cholic acid.

6. The application according to any one of claims 1-5, characterized in that, The active ingredients of the traditional Chinese medicine composition are prepared by the following method: taking the Chinese gall, Japanese pagoda tree flower, Ampelopsis japonica, Selaginella tamariscina and Sanguisorba officinalis, mixing them, soaking them in water, then decocting, combining the decocted medicinal liquids, and then adding the cholic acid and mixing evenly to obtain.

7. The application according to claim 6, wherein Decoct 2-4 times. The water addition amount for the first decoction is 6-8 times the total weight of the medicinal materials, and the water addition amount for each subsequent decoction is 4-6 times the total weight of the medicinal materials; and / or, the decoction time for each time is 20 min-40 min; and / or, the soaking time is 20 min-40 min.

8. The application according to claim 7, wherein Decoct 3 times. The water addition amount for the first decoction is 7 times the total weight of the medicinal materials, and the water addition amount for each subsequent decoction is 5 times the total weight of the medicinal materials; and / or, the decoction time for each time is 25 min-35 min; and / or, the soaking time is 25 min-35 min.

9. The application according to any one of claims 1 to 5, characterized in that, The dosage form of the anti-colorectal cancer drug is tablet, injection, capsule, granule, powder, mixture, emulsion, decoction or suspension.

10. The application according to any one of claims 1-5, characterized in that, The drug can reduce the levels of inflammatory factors in colon tissue and serum tumor markers.

Images