Composition for improving ackermann abundance and inhibiting tumor growth and application

Abstract

The invention discloses a composition for improving ackermann abundance and inhibiting tumor growth and application, and belongs to the technical field of biological medicine. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 10-20 parts of pinellia ternate or 5-15 parts of hawthorn, 10-20 parts of pericarpium citri reticulatae, 5-15 parts of poria cocos, 2-11 parts of honey-fried licorice root, 2-11 parts of ginger and 2-11 parts of dark plum. The composition provided by the invention affects the intestinal flora structure and improves the abundance of Ackerman bacteria in the intestinal tract; the number of goblet cells and the expression of intestinal tight junction proteins ZO-1 and Occludin are increased, and the barrier function of the intestinal tract is improved; the number of CD8 < + > T cells and GZMB < + > cells in a tumor microenvironment is increased, and the effect of inhibiting tumor growth is achieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine. Background Art

[0002] The gut-liver axis, established through the portal vein, plays an important role in the metabolism of nutrients, the transformation of bacterial metabolites, and the regulation of the immune system. The development of liver cancer is associated with dysbiosis of the intestinal flora, which is associated with damaged intestinal barriers, reduced flora diversity, increased harmful bacteria, and decreased beneficial bacteria. The intestinal flora influences the development of liver cancer by promoting anti-tumor immunity and regulating the tumor microenvironment, playing a key role in tumor progression and prognosis. The progression of liver cancer is accompanied by changes in the structure of the intestinal flora. Improving the flora structure of liver cancer patients provides new ideas for the diagnosis and treatment of liver cancer patients. Summary of the Invention

[0003] The technical problem to be solved by the present invention is to provide a composition and its application for increasing the abundance of Akkermansia to inhibit tumor growth. The Chinese medicine prescription of the present invention affects the structure of intestinal flora, increases the abundance of Akkermansia, enhances the intestinal barrier function, increases the activation of CD8 + T number, playing the role of inhibiting tumor growth.

[0004] In order to solve the above technical problems, the technical solution adopted by the present invention is: A composition for increasing the abundance of Akkermansia and inhibiting tumor growth (composition one) comprises the following substances by weight: 10-20 parts of pinellia, 10-20 parts of dried tangerine peel, 5-15 parts of poria, 2-11 parts of roasted licorice, 2-11 parts of ginger, and 2-11 parts of black plum.

[0005] The preferred composition comprises the following raw materials by weight: 12-18 parts of pinellia tuber, 12-18 parts of dried tangerine peel, 6-12 parts of poria cocos, 3-9 parts of roasted licorice root, 4-10 parts of ginger, and 5-11 parts of black plum.

[0006] The preferred composition comprises the following raw materials by weight: 15 parts of pinellia tuber, 15 parts of dried tangerine peel, 9 parts of poria cocos, 6 parts of roasted licorice root, 7 parts of ginger, and 8 parts of black plum.

[0007] A composition for increasing the abundance of Akkermansia and inhibiting tumor growth (composition 2) is composed of the following raw materials by weight: 5-15 parts of hawthorn, 10-20 parts of tangerine peel, 5-15 parts of poria, 2-11 parts of roasted licorice, 2-11 parts of ginger, and 2-11 parts of black plum.

[0008] The preferred raw materials of the second composition are composed of the following substances by weight: 7-13 parts of hawthorn, 12-18 parts of dried tangerine peel, 6-12 parts of poria, 3-9 parts of roasted licorice, 4-10 parts of ginger, and 5-11 parts of black plum.

[0009] The preferred raw materials of the second composition are composed of the following substances by weight: 10 parts of hawthorn, 15 parts of dried tangerine peel, 9 parts of poria, 6 parts of roasted licorice, 7 parts of ginger, and 8 parts of black plum.

[0010] The present invention also provides the use of the composition in preparing a preparation for inhibiting tumor growth; after administration of the preparation, CD8 + T cells and GzmB + The number of cells increased and the accumulation of lipid droplets in the tumor microenvironment decreased.

[0011] The present invention also provides the use of the composition in preparing a preparation that affects the structure of intestinal flora; after administration of the preparation, the relative abundance of Akkermansia, Dublerella, Dublerella, and Escherichia coli in the intestine is increased.

[0012] The present invention also provides the use of the composition in preparing a preparation for enhancing intestinal barrier function; after administration of the preparation, the number of goblet cells increases and the expression of tight junction proteins ZO-1 and Occludin is improved.

[0013] The present invention also provides the use of the second composition in preparing a preparation for inhibiting tumor growth.

[0014] The present invention also provides a combination of composition 2 and cisplatin for use in preparing a preparation for inhibiting tumor growth; after administration of the preparation, CD8 + T cell numbers and GzMB + The number of cells increases.

[0015] The present invention also provides the use of the second composition in preparing a preparation for influencing the structure of intestinal flora.

[0016] The present invention also provides the combination of the second composition and cisplatin for use in a preparation for influencing the structure of intestinal flora; after administration of the preparation, the abundance of Akkermansia genus in the intestinal tract is increased.

[0017] The present invention also provides the use of the second composition in the preparation of a preparation for enhancing intestinal barrier function.

[0018] The present invention also provides a combination of the second composition and cisplatin for use in a preparation for enhancing intestinal barrier function; after administration of the preparation, the number of goblet cells increases and the expression of tight junction proteins ZO-1 and Occludin is enhanced.

[0019] The beneficial effects of adopting the above technical solution are: The composition of the present invention affects the structure of intestinal flora, increases the abundance of Akkermansia, enhances intestinal barrier function, reduces lipid droplet accumulation in tumor microenvironment, and increases activation of CD8 + T number, playing the role of inhibiting tumor growth.

[0020] The combined use of the second composition of the present invention and cisplatin affects the intestinal flora structure, increases the abundance of Akkermansia, enhances the intestinal barrier function, and increases the activation of CD8 + T number, playing the role of inhibiting tumor growth. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a graph showing the effect of the composition of Example 1 of the present invention on inhibiting the growth of liver cancer.

[0022] Figure 2 This is a graph showing the effect of increasing the abundance of Akkermansia on the bacterial community structure of the composition of Example 1.

[0023] Figure 3 This is a graph showing the effect of the composition of Example 1 on the intestinal barrier function.

[0024] Figure 4 The composition of Example 1 is a pair of CD8 + T cells and GzmB + The number of cells and lipid droplet deposition affect the graph.

[0025] Figure 5 This is a graph showing the effects of Composition 2, Composition 2+DDP, and DDP in inhibiting liver cancer growth in Example 2.

[0026] Figure 6 This is a graph showing the effect of Composition 2+DDP on the bacterial community structure and the abundance of Akkermansia in Example 2.

[0027] Figure 7 This is a graph showing the effect of Composition 2+DDP on intestinal barrier function in Example 2.

[0028] Figure 8 Composition 2 of Example 2 acts on CD8 + T cells and GzmB + Cell number effect diagram. DETAILED DESCRIPTION Example 1

[0029] A subcutaneous hepatocellular carcinoma (HCC) xenograft model was established in 5-week-old C57BL / 6 male mice using Hepa1-6 cells. After tumor formation, the mice were randomly divided into two groups and gavaged with sterile water or Composition 1, respectively. Tumor volume and mouse body weight were measured and recorded every three days. Feces were collected for 16S rRNA sequencing 14 days after intervention. Colon and tumor tissues were obtained and fixed with 4% paraformaldehyde 15 days after intervention for subsequent experiments.

[0030] Experimental methods: (1) Animal modeling and grouping The animals were housed in a SPF facility at a constant temperature (22-24°C) with a 12 h / 12 h light cycle. Ten 5-week-old C57BL / 6 males (average weight 18 g) were selected. 0.1 ml of Hepa1-6 cell suspension (1 × 10 cells) was injected subcutaneously into the right hind leg. 7 Tumor-bearing mice were divided into a control group (Control group) and a combination group (ECD group). The control group was given 0.1 mL / day of sterile water by gavage. The combination group was given 7.8 g / kg / day of the drug solution by gavage.

[0031] (2) Preparation of Chinese medicine prescriptions The adult dosage of 15 parts of Pinellia, 15 parts of Tangerine Peel, 9 parts of Poria, 6 parts of Roasted Licorice, 7 parts of Ginger, and 8 parts of Prunus mume was converted to the mouse dosage based on the body surface area algorithm, i.e., 1.95 g / kg of Pinellia, 1.95 g / kg of Tangerine Peel, 1.17 g / kg of Poria, 0.78 g / kg of Roasted Licorice, 0.91 g / kg of Ginger, and 1.04 g / kg of Prunus mume. The Chinese herbal medicine solution was prepared by boiling ginger, tangerine peel, Poria, Roasted Licorice, ginger, and Prunus mume according to the above weight ratios and concentrating to a gavage dose of 0.1 mL per mouse. (3) Measurement of transplanted tumor volume changes The length and width of the tumor were measured with a vernier caliper every 3 days, and the formula The tumor volume was calculated (V represents tumor volume, L represents long diameter, and I represents short diameter), and the tumor growth curve was drawn.

[0032] (4) Mouse weight measurement The mice were weighed every 3 days using an electronic scale and the weight was recorded to draw a weight curve changing over time.

[0033] (5) 16S rRNA sequencing After 14 days of intervention, the feces of mice in each group were collected. The microbial diversity in mice was analyzed by sequencing by Shanghai Paisono Biotechnology Co., Ltd. The rarefaction curves were analyzed and drawn using the Gene Cloud website. diversity and species differences, etc.

[0034] (6) Paraffin section and H&E staining After being sliced into 3-5 μm sections using an automatic embedding machine, the sections were flattened in 40°C water and removed using a glass slide. After drying the sections on a slide dryer, they were placed in a 67°C oven to dissolve the wax for 1 hour before removal. After the sections cooled to room temperature, the staining procedure began. Conventional dewaxing and dehydration were performed. Staining consisted of hematoxylin staining, rinsing with tap water, differentiation solution, bluing with tap water, staining with eosin, and rinsing with tap water to remove excess stain. Dehydration was then performed in standard alcohol, followed by xylene clearing, and finally mounting with neutral gum. Tumor histopathological changes were observed using a light microscope.

[0035] (7) Colon PAS staining Colonic paraffin sections were sectioned and stained according to the instructions of the PAS kit (R20526, Shanghai Yuanye Biotechnology Co., Ltd.). The morphology was observed under an optical microscope and photographed.

[0036] (8) Immunohistochemical analysis The sections were dewaxed and hydrated for antigen retrieval. After adding 3% hydrogen peroxide to eliminate the influence of endogenous enzymes, the sections were blocked and incubated with primary antibodies ZO-1 (ab216880, Abcam), Occludin (27260-1-AP, Proteintech), and CD8 + T (D4W2Z, Cell Signaling Technology), GZMB (D2H2F, Cell Signaling Technology), PLIN2 (15294-1-AP, Proteintech) antibodies were used, followed by incubation with secondary antibodies; detailed staining steps were performed according to the instructions of the rabbit SP kit; DAB color development steps were performed according to the instructions of the DAB kit; hematoxylin was used to stain the nuclei, differentiate, return to blue, dehydrate with graded alcohols, clear with xylene, and mount with neutral gum; the results were observed under an optical microscope and photographed. ZO-1, Occludin, and CD8 were detected in this way respectively. + Expression levels of T, GZMB, and PLIN2 in tissues.

[0037] (9) Oil Red O staining Prepare frozen tissue sections: Soak fresh tissue stored in paraformaldehyde in a 30% sucrose solution for 48 hours, then embed the tissue using a cryostat. Cut frozen sections onto slides using a cryostat and store at -20°C.

[0038] Modified Oil Red O staining (G1261, Solarbio): Frozen sections were washed with 60% isopropanol for 30 s, stained with Oil Red O staining solution for 15 min, washed with 60% isopropanol for 30 s, counterstained with hematoxylin for 90 s, washed with water for 10 min, washed with distilled water for 3 min, mounted with glycerol gelatin, and observed under a microscope after drying.

[0039] Experimental results: (1) ECD inhibits liver cancer growth As attached Figure 1 As shown, there was no significant difference in body weight between the ECD group (19.22±1.205 g) and the Control group (19.06±0.8678 g) ( P >0.05). Compared with the Control group (287.3±218.1 mm 3 ) compared with the ECD group (45.25±42.72mm 3 ) tumor volume reduction ( P <0.05). These results indicate that ECD inhibits HCC growth without affecting body weight.

[0040] (2) ECD affects the bacterial community structure and increases the abundance of Akkermansia We used 16S rRNA sequencing of mouse feces to further examine the effect of ECD on the intestinal microbiota composition of mice with subcutaneous HCC transplants. The rarefaction curve gradually stabilized with the increase in sample number and no longer increased, indicating that the sequencing depth had reached a sufficient level and the sequencing data was reliable (see attached). Figure 2 A).

[0041] Principal coordinate analysis (PCoA) was used to examine the changes in gut microbiome separation between different groups of mice. Our analysis showed that there were differences in microbial community composition between the Control and ECD groups (e.g., Figure 2 B).

[0042] In addition, to comprehensively assess the overall microbial diversity, we evaluated The Goods_coverage index of both groups was close to 1, indicating that the sequencing results can represent the true situation of the samples. Compared with the Control group, the Shannon, Simpson, and Chao1 indices of the ECD group showed no significant difference ( P >0.05) (if attached Figure 2 C), indicating that the ECD group and the Control group had similar microbial diversity, microbial richness, and species richness.

[0043] In order to study the changes in the intestinal flora of mice, we analyzed the species abundance of the intestinal flora of each group of mice at different levels. First, the species composition of the intestinal flora of mice was analyzed at the phylum level. Although Firmicutes and Bacteroidota were still the dominant phyla under ECD intervention, the relative abundance of Firmicutes decreased, and the relative abundance of Bacteroidetes did not change. It is worth noting that the relative abundance of Verrucomicrobiota and Proteobacteria increased (as shown in Figure 2D). At the genus level, the relative abundance of Akkermansia, Duncanilla, Dubosiella, and Escherichia increased after ECD intervention (as shown in Figure 2D). Figure 2 E, F). Therefore, we believe that ECD affects the structure of the intestinal flora and increases the relative abundance of Akkermansia.

[0044] (3) ECD enhances intestinal barrier function After identifying the changes in intestinal flora composition, we performed H&E and PAS staining on the mouse colon. The results showed that the colon structure of the control group and the ECD group was intact, with goblet cells arranged tightly and no obvious pathological damage (such as attached Figure 3 A). Compared with the control group, the crypt depth decreased and the number of goblet cells increased after ECD treatment (see attached Figure 3 A). This indicates that ECD increases the maturation rate of mouse colonic villus epithelial cells and mucus secretion, and strengthens the barrier function of the intestinal mucosa. The expression levels of ZO-1 and Occludin in the mouse colon increased in the ECD group (see attached). Figure 3 B). This indicates that ECD increases the number of goblet cells, enhances the expression of tight junction proteins ZO-1 and occludin, and strengthens intestinal barrier function.

[0045] (4) ECD reduces lipid droplet deposition and increases activated CD8 + T quantity ECD increases CD8 + T cells and GzmB + The number of cells (e.g. Figure 4 B). At the same time, we found that after ECD intervention, the accumulation of lipid droplets in tumor tissues decreased, and the expression of the fat differentiation-related protein PLIN2 decreased, which also reflected that ECD reduced fat accumulation in tumor tissues (see attached Figure 4 C). Therefore, ECD reduces the accumulation of lipid droplets in liver cancer tissue and increases the activation of CD8 + T quantity The above results show that the composition of this embodiment affects the structure of intestinal flora, increases the abundance of Akkermansia, enhances intestinal barrier function, reduces the accumulation of lipid droplets in the tumor microenvironment, and increases the activation of CD8 + T number, playing the role of inhibiting tumor growth. Example 2

[0046] A subcutaneous hepatocellular carcinoma (HCC) xenograft model was established in 5-week-old C57BL / 6 male mice using Hepa1-6 cells. After tumor formation, the mice were randomly divided into two groups and treated with sterile water, combination II (HCF), combination II + cisplatin (HCF + DDP), or cisplatin (HCF). Tumor volume and mouse body weight were measured and recorded every three days. Feces were collected for 16S rRNA sequencing on day 14 of the intervention. Colon and tumor tissues were obtained on day 15 of the intervention and fixed with 4% paraformaldehyde for subsequent experiments.

[0047] Experimental methods: (1) Animal modeling and grouping Animals were housed in a SPF facility at a constant temperature (22-24°C) with a 12 h / 12 h light cycle. Twenty 5-week-old C57BL / 6 males (average weight 18 g) were selected. 0.1 ml of Hepa1-6 cell suspension (1 × 10 cells) was injected subcutaneously into the right hind leg. 7 Tumor-bearing mice were divided into a control group (Control group), a Composition II group (HCF group), a Composition II + Cisplatin group (HCF + DDP group), and a Cisplatin group (DDP group). The control group was treated with 0.1 mL / day of sterile water by oral gavage, the Composition II group was treated with 7.15 g / kg / day of the drug solution by oral gavage, the Composition II + Cisplatin group was treated with 2 mg / kg every two days by intraperitoneal injection in addition to the Composition II group treatment, and the Cisplatin group was treated with 2 mg / kg every two days by intraperitoneal injection.

[0048] (2) Preparation of Chinese medicine prescriptions The adult dosage of 10 parts hawthorn, 15 parts dried tangerine peel, 9 parts poria, 6 parts roasted licorice root, 7 parts ginger, and 8 parts black plum was converted to the mouse dosage based on the body surface area algorithm, i.e., 1.3 g / kg hawthorn, 1.95 g / kg dried tangerine peel, 1.17 g / kg poria, 0.78 g / kg roasted licorice root, 0.91 g / kg ginger, and 1.04 g / kg black plum. The Chinese herbal medicine solution was prepared by boiling hawthorn, dried tangerine peel, poria, roasted licorice root, ginger, and black plum according to the above weight ratios and concentrating to a gavage dose of 0.1 mL per mouse. (3) Measurement of transplanted tumor volume changes Same as Example 1

[0049] (4) Mouse weight measurement Same as Example 1

[0050] (5) 16S rRNA sequencing Same as Example 1

[0051] (6) Paraffin section and H&E staining Same as Example 1

[0052] (7) Colon PAS staining Same as Example 1

[0053] (8) Immunohistochemical analysis The sections were dewaxed and hydrated for antigen retrieval. After adding 3% hydrogen peroxide to eliminate the influence of endogenous enzymes, the sections were blocked and incubated with primary antibodies ZO-1 (ab216880, Abcam), Occludin (27260-1-AP, Proteintech), and CD8 + T (D4W2Z, Cell Signaling Technology), GZMB (D2H2F, Cell Signaling Technology) antibodies, and then incubated with secondary antibodies; detailed staining steps were carried out according to the instructions of the rabbit SP kit; DAB color development steps were carried out according to the instructions of the DAB kit; hematoxylin nuclear staining, differentiation, blueing, gradient alcohol dehydration, xylene clearing, neutral gum sealing; the results were observed under an optical microscope and photographed. ZO-1, Occludin, and CD8 were detected in this way respectively. + Expression levels of T, GZMB, and PLIN2 in tissues.

[0054] Experimental results: (1) HCF, HCF+DDP, and DDP inhibit liver cancer growth As attached Figure 5 As shown, there was no significant difference in body weight between the HCF group (18.44±0.7197 g) and the Control group (19.20±0.8775 g) ( P >0.05), but the body weight of the HCF+DDP group (15.98±1.902 g) and the DDP group (15.72±1.361 g) was lower than that of the Control group ( P <0.05). There was no significant difference in body weight between the HCF+DDP group and the DDP group ( P >0.05). This indicates that DDP intervention reduced the body weight of mice, but HCF had no effect on the body weight of mice.

[0055] We analyzed the tumor volume on the day of sampling and found that compared with the Control group (285.4±211.4 mm 3) compared with the HCF group (63.62±69.26 mm 3 ), HCF+DDP group (9.939±3.751 mm 3 )、DDP(67.74±53.91 mm 3 ) tumor volume reduction ( P <0.05). This indicates that HCF, HCF+DDP, and DDP can all inhibit tumor growth.

[0056] (2) HCF+DDP affects the bacterial community structure and increases the abundance of Akkermansia We used 16S rRNA sequencing of mouse feces to further examine the effects of HCF, HCF+DDP, and DDP on the intestinal flora composition of mice with subcutaneous HCC transplants. The rarefaction curve gradually stabilized with the increase in sample number and no longer increased (see Appendix Figure 6 A), indicating that the sequencing depth has reached a sufficient level and the sequencing data are reliable and can be used for subsequent experiments.

[0057] Principal coordinate analysis (PCoA) was used to examine the separation of intestinal microbial components between different groups of mice. Our analysis showed that there were differences in microbial community composition between the Control group, HCF group, and HCF+DDP group, but there was no significant difference in microbial community composition between the DDP group and the other three groups (see attached). Figure 6 B).

[0058] In addition, to comprehensively assess the overall microbial diversity, we evaluated Diversity. The Goods_coverage index between the groups was close to 1, indicating that the sequencing results can represent the actual situation of the samples. From the Shannon, Simpson, and Chao1 index analysis, it was found that the HCF and Control groups had similar microbial diversity, richness, and species richness. After DDP intervention, the microbial diversity, richness, and species richness were reduced. HCF can improve the microbial diversity and richness reduced by DDP (such as the attached Figure 6 C).

[0059] To study the changes in the intestinal flora of mice, we analyzed the species abundance of the intestinal flora of each group of mice at different levels. First, the species composition of the intestinal flora of mice was analyzed at the phylum level. Although Firmicutes and Bacteroidota were still the dominant phyla under the intervention of HCF, HCF+DDP and DDP, the F / B value of the HCF group (0.644426), the F / B value of the HCF+DDP group (0.420088) and the F / B value of the DDP group (0.45731) were lower than the F / B value of the Control group (0.775056), indicating that the structure of the flora had changed (such as the presence of Figure 6D). It is worth noting that the relative abundance of Verrucomicrobiota and Proteobacteria increased in the HCF+DDP group, and the relative abundance of Actinobacteriota increased in the DDP group.

[0060] At the genus level, CAG-485 was the dominant genus in the HCF group, Ligilactobacillus, Parasutterella, Akkermansia, Duncanilla, and Bacteroides_H were the dominant genera in the HCF+DDP group, and Bifidobacterium, Lactobacillus, Bacteroides_H, and Duncanilla were the dominant genera in the DDP group (see attached). Figure 6 E, F). The relative abundance of Akkermansia, a genus of interest, was highest in the HCF+DDP group, followed by the DDP group. Therefore, we conclude that both experimental groups affected the structure of the intestinal microbiota, with the HCF+DDP group significantly increasing the relative abundance of Akkermansia.

[0061] (3) HCF+DDP enhances intestinal barrier function After identifying the changes in intestinal flora composition, we performed H&E and PAS staining on the mouse colon. The results showed that the colon structures of the control group, HCF group, HCF+DDP group, and DDP group were intact, with goblet cells arranged tightly and no obvious pathological damage (such as attached Figure 7 A).

[0062] PAS staining showed that the number of goblet cells in the HCF group and HCF+DDP group increased compared with the Control group, while there was no significant difference in the DDP group. Figure 7 A). This indicates that HCF increases goblet cell number, while DDP has no effect on goblet cell number. However, the combined use of HCF and DDP resulted in a more significant increase in goblet cell numbers than either HCF or DDP alone. Therefore, HCF + DDP increases the maturation rate of colonic villus epithelial cells and mucus secretion in mice, strengthening the intestinal mucosal barrier function.

[0063] When the intestinal barrier function increases, the expression of ZO-1 and Occludin also increases. Compared with the control group, the expression levels of ZO-1 and Occludin in the colon of mice were reduced in the HCF and DDP groups, but increased in the HCF+DDP group. Compared with the HCF and DDP groups, the HCF+DDP group significantly increased the expression of ZO-1 and Occludin (see attached figure). Figure 7B). This suggests that HCF+DDP increases intestinal tight junctions, while HCF and DDP decrease them. This further demonstrates that while the HCF, HCF+DDP, and DDP groups all inhibited tumor growth, the HCF+DDP group was more effective than either HCF or DDP alone in affecting microbial structure, increasing the abundance of Akkermansia, and enhancing intestinal barrier function.

[0064] (4) HCF+DDP increases activated CD8 + T quantity Compared with the control group, HCF and HCF+DDP intervention increased the activation of CD8 in the tumor microenvironment in tumor tissues. + The number of T cells increased in the DDP group, but only CD8 + Compared with the HCF and DDP groups, HCF+DDP increased the number of activated CD8 T cells in the tumor microenvironment. + The number of T cells (see Figure 8B) indicates that HCF and HCF+DDP increase the activation of CD8 T cells in the tumor microenvironment. + The number of T cells was significantly increased by HCF+DDP, but DDP could not increase the number of activated CD8 T cells in the tumor microenvironment. + T cell count.

[0065] The above results show that the combined use of the second composition of this embodiment and cisplatin affects the intestinal flora structure, increases the abundance of Akkermansia, enhances the intestinal barrier function, and increases the activation of CD8 + T number, playing the role of inhibiting tumor growth.

Claims

1. A composition for increasing the abundance of Akkermansia and inhibiting tumor growth, characterized in that: The raw materials are composed of the following substances by weight: 10-20 parts of pinellia or 5-15 parts of hawthorn, 10-20 parts of dried tangerine peel, 5-15 parts of poria, 2-11 parts of roasted liquorice, 2-11 parts of ginger and 2-11 parts of black plum.

2. The composition according to claim 1, wherein: The raw materials are composed of the following substances by weight: 12-18 parts of pinellia or 7-13 parts of hawthorn, 12-18 parts of dried tangerine peel, 6-12 parts of poria, 3-9 parts of roasted liquorice, 4-10 parts of ginger and 5-11 parts of black plum.

3. The composition according to claim 2, characterized in that: The raw materials are composed of the following substances by weight: 15 parts of pinellia tuber, 15 parts of dried tangerine peel, 9 parts of poria, 6 parts of roasted liquorice root, 7 parts of ginger and 8 parts of black plum.

4. The composition according to claim 2, wherein: The raw materials are composed of the following substances by weight: 10 parts of hawthorn, 15 parts of dried tangerine peel, 9 parts of poria, 6 parts of roasted licorice, 7 parts of ginger and 8 parts of black plum.

5. Use of the composition according to any one of claims 1 to 3 in the preparation of a preparation for inhibiting tumor growth.

6. Use of the composition according to any one of claims 1 to 3 in the preparation of a preparation for influencing the structure of intestinal flora.

7. Use of the composition according to any one of claims 1 to 3 in the preparation of a preparation for enhancing intestinal barrier function.

8. Use of the composition according to any one of claims 1, 2 and 4 in the preparation of a preparation for inhibiting tumor growth.

9. Use of the composition according to any one of claims 1, 2 and 4 in the preparation of a preparation for influencing the structure of intestinal flora.

10. Use of the composition according to any one of claims 1, 2 and 4 in the preparation of a preparation for enhancing intestinal barrier function.

Images