Application of fructus cinnamoii in preparation of medicine for treating inflammatory bowel disease

Abstract

The invention belongs to the technical field of medicines, and particularly relates to application of fructus cinnamoii in preparation of medicines for treating inflammatory bowel diseases. According to the application, it is disclosed for the first time that the fructus cinnamoii oil and the fructus cinnamoii aqueous extract can relieve weight loss of IBD mice, reduce the DAI score, and significantly recover colon length (Plt; 0.05), the pathological damage of colon tissues is reduced, and the contents (Plt; Pt) of pro-inflammatory cytokines TNF-alpha, IL-6 and IL-1beta in colon are remarkably reduced; 0.05) of the method. Alpha diversity and beta diversity analysis results show that the litsea coreana oil and aqueous extract can change the overall flora structure of the DSS-induced IBD mouse, so that the community composition of the intestinal flora of the mouse approaches to a normal group. The relative abundance of the intestinal flora at the phylum level, the family level and the genus level is further analyzed, and it is found that the fructus cinnamoii oil and the fructus cinnamoii water extract can influence the composition of the intestinal flora of the mouse in the pathological state. By measuring the contents of six short-chain fatty acids in the excrement of each group of mice, the results show that the contents of acetic acid and butyric acid (Plt; 0.05), which indicates that the fructus cinnamoii oil and the fructus cinnamoii aqueous extract have prevention and treatment effects on the inflammatory bowel disease. Therefore, the fructus cinnamoii has a good application prospect in the aspect of preparing the medicines related to the inflammatory bowel disease.

Description

Technical Field

[0001] The present application belongs to the field of medical technology, and specifically relates to the application of Litsea cubeba in the preparation of drugs for inflammatory bowel disease. Background Art

[0002] Inflammatory bowel disease (IBD) is a nonspecific intestinal disease characterized by chronic inflammation and ulcer formation. IBD has been listed as one of the difficult-to-cure diseases in modern times by the World Health Organization because of its easy recurrence and difficulty in cure. It is estimated that by 2025, the number of IBD patients in my country will exceed 1.5 million, ranking first in Asia, and becoming one of the most common diseases of the digestive system in my country. To date, the etiology and pathogenesis of IBD are still unclear, but it is now generally believed that intestinal flora plays an important role in the occurrence and development of enteritis. The intestinal flora maintains a dynamic balance with the host immune system, tolerates the presence of normal intestinal bacteria, and inhibits the proliferation and expansion of pathogenic microorganisms. Intestinal dysbiosis increases the proportion of harmful bacteria in the intestine, releases enterotoxins, increases intestinal permeability, induces the production of immunosuppressive proteins, leads to immune dysfunction, destroys intestinal epithelial cells, affects energy metabolism, and causes intestinal inflammation.

[0003] At present, the drugs used in clinical treatment of IBD mainly include aminosalicylic acid preparations, glucocorticoids, immune and biological inhibitors, etc., which have the characteristics of rapid onset and obvious effects in the short term, but the long-term efficacy is not ideal and the side effects are too large. As a traditional advantageous resource in my country, Chinese medicine has the characteristics of multiple components, multiple pathways, multiple targets and synergistic effects, which give it unique advantages in preventing and treating diseases. In the process of searching for new drugs to treat IBD, Chinese medicine and Chinese medicine extracts have received widespread attention and achieved many results.

[0004] As a common medicine for ethnic minorities in Guizhou, Litsea cubeba is included in the 2019 edition of the Quality Standards for Traditional Chinese and Ethnic Chinese Medicinal Materials in Guizhou Province. It is mainly used to treat stomachache, abdominal pain, chest pain, etc. So far, it has not been reported that Litsea cubeba has the effect of preventing and treating inflammatory bowel disease. Summary of the invention

[0005] In order to solve the above problems, the present application provides the use of Litsea cubeba in the preparation of inflammatory bowel disease drugs. Specifically, the application is achieved through the following technical solutions:

[0006] Application of Litsea cubeba in preparing medicine for inflammatory bowel disease.

[0007] Furthermore, the Litsea cubeba includes products of Litsea cubeba, such as Litsea cubeba oil and Litsea cubeba water extract.

[0008] Furthermore, the inflammatory bowel disease drug is a drug for preventing or treating inflammatory bowel disease.

[0009] Furthermore, the inflammatory bowel disease drug is a drug that reduces the symptoms of inflammatory bowel disease such as blood in the stool and loose stools.

[0010] Furthermore, the inflammatory bowel disease drug is a drug that improves the symptoms of shortened colon length.

[0011] Furthermore, the inflammatory bowel disease drug is a drug that reverses pathological damage to colon tissue.

[0012] Furthermore, the inflammatory bowel disease drug is a drug that reverses changes in inflammatory responses, inhibits inflammatory responses, and reduces the levels of TNF-α, IL-6, and IL-1β.

[0013] Furthermore, the inflammatory bowel disease drug is a drug that adjusts the intestinal flora structure toward normal.

[0014] In summary, the beneficial effects of the present application are as follows: The present application discloses for the first time that Litsea cubeba oil and Litsea cubeba water extract can alleviate weight loss in IBD mice, reduce DAI scores, significantly restore colon length (P<0.05), reduce colon tissue pathological damage, and significantly reduce the content of pro-inflammatory cytokines TNF-α, IL-6 and IL-1β in the colon (P<0.05). The results of α diversity and β diversity analysis found that Litsea cubeba oil and water extract can change the overall flora structure of DSS-induced IBD mice, making the community composition of the mouse intestinal flora closer to that of the normal group. Further analysis of the relative abundance of intestinal flora at the phylum, family, and genus levels found that Litsea cubeba oil and Litsea cubeba water extract can affect the composition of the intestinal flora of mice under pathological conditions. By measuring the contents of 6 short-chain fatty acids in the feces of mice in each group, it was found that the contents of acetic acid and butyric acid were significantly increased after intervention with Litsea cubeba oil and water extract (P<0.05), indicating that Litsea cubeba oil and Litsea cubeba water extract have preventive and therapeutic effects on inflammatory bowel disease. This shows that Litsea cubeba has good application prospects in the preparation of drugs related to inflammatory bowel disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 The modeling and dosing regimen of the IBD model in mice include dextran sulfate sodium: DSS; sulfasalazine: SASP.

[0016] Figure 2 This is the flowchart of 16SrDNA high-throughput sequencing.

[0017] Figure 3 The effects of CMO and CMW on the physical signs of IBD model mice (n=10), where A: body weight change curve; B: DAI score.

[0018] Figure 4The effect of CMO and CMW on the colon length of mice (n=10), among which compared with the CON group, ### P<0.001; compared with the MOD group, * P<0.05, ** P<0.01, *** P<0.001.

[0019] Figure 5 Results of colon histopathological examination.

[0020] Figure 6 The effect of CMO and CMW on TNF-α in the colon of mice (n=10), compared with the CON group, ### P<0.001; compared with the MOD group, ** P<0.01, *** P<0.001.

[0021] Figure 7 The effect of CMO and CMW on IL-6 in the colon of mice (n=10), compared with the CON group, ### P<0.001; compared with the MOD group, ** P<0.01, *** P<0.001.

[0022] Figure 8 The effect of CMO and CMW on IL-1β in the colon of mice (n=10), compared with the CON group, ### P<0.001; compared with the MOD group, *** P<0.001.

[0023] Fig. 9 Effects of CMO and CMW on the α-diversity of intestinal flora in mice.

[0024] Fig.10 Effects of CMO and CMW on the β-diversity of intestinal flora in mice.

[0025] Fig.11 Effects of CMO and CMW on the phylum levels of intestinal flora in mice.

[0026] Fig.12 Effects of CMO and CMW on the family levels of intestinal flora in mice.

[0027] Fig.13 The effects of CMO and CMW on the genus levels of intestinal flora in mice.

[0028] Fig.14 This is the result of short-chain fatty acid content determination. DETAILED DESCRIPTION

[0029] The specific implementation methods of the present application are further described in detail below, but the present application is not limited to these implementation methods. Any improvement or substitution based on the basic spirit of the present embodiment still falls within the scope of protection required by the claims of the present application.

[0030] Example 1

[0031] 1 Experimental Materials

[0032] 1.1 Main drugs and reagents

[0033] Dextran sulfate sodium (S7980) Sulfasalazine (Y23A9C59570) from MPBiomedicals, USA Fecal occult blood (OB) reagent (B221101) from Shanghai Yuanye Biotechnology Co., Ltd. Mouse TNF-α kit (ZC-39024) from Zhuhai Beso Biotechnology Co., Ltd. Mouse IL-1β kit (ZC-37974) from Shanghai Zhuocai Biotechnology Co., Ltd. Mouse IL-6 kit (ZC-37988) from Shanghai Zhuocai Biotechnology Co., Ltd. 1.2 Experimental animals

[0034] 100 C57BL / 6 male mice, 8 weeks old, weighing 22±2g, were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd., license number SCXK (Beijing) 2019-0010. The mice were kept at a temperature of 20-25°C, a relative humidity of 40-70%, free access to water and food, and were adaptively fed for 1 week.

[0035] 2 Methods

[0036] 2.1 Preparation of Litsea cubeba oil

[0037] Take an appropriate amount of Litsea cubeba, crush it, add 8 times the amount of water to soak for 30 minutes, extract for 5 hours, collect the volatile oil, add an appropriate amount of anhydrous sodium sulfate, seal overnight, centrifuge at 8000r / min for 10 minutes the next day, collect the upper oil layer to obtain Litsea cubeba oil (CMO).

[0038] 2.2 Preparation of Litsea cubeba aqueous extract

[0039] Take an appropriate amount of Litsea cubeba, crush it, add 8 times the amount of water to soak for 30 minutes, reflux extraction for 1 hour, twice in total, combine the filtrate, concentrate and dry to obtain the water extract (CMW).

[0040] 2.3 Solution preparation

[0041] Preparation of Litsea cubeba oil: Weigh an appropriate amount of Litsea cubeba oil, place it in a beaker, add 0.5% CMC-Na, ultrasonically treat it in an ice-water bath for 10 minutes to dissolve it, and prepare it for use before intragastric administration.

[0042] Preparation of Litsea cubeba water extract: Take an appropriate amount of Litsea cubeba water extract, place it in a beaker, add 0.5% CMC-Na, ultrasonicate it in an ice water bath for 10 minutes to dissolve it, and prepare it before intragastric administration.

[0043] Preparation of dextran sodium sulfate solution: Weigh an appropriate amount of dextran sodium sulfate, place it in a beaker, add an appropriate amount of distilled water to dissolve it, and prepare a 2% dextran sodium sulfate solution.

[0044] Preparation of sulfasalazine solution: Weigh an appropriate amount of sulfasalazine, place in a beaker, add 0.5% CMC-Na, ultrasonicate for 10 minutes to dissolve, and prepare and use immediately before oral administration.

[0045] 2.4 Animal modeling and drug administration process

[0046] C57BL / 6 mice were randomly divided into 9 groups, with 10 mice in each group, namely normal group (CON), model group (MOD), positive drug group (sulfasalazine, SASP), low-, medium- and high-dose groups of Litsea cubeba oil (CMOL, CMOM, CMOH), and low-, medium- and high-dose groups of Litsea cubeba water extract (CMWL, CMWM, CMWH). The specific dosing schedule is shown in Figure 1 The normal group freely drank distilled water every day, and the mice in other groups freely drank 2% dextran sulfate sodium (DSS) aqueous solution for 7 consecutive days to induce the IBD model. Drug intervention was given from the first day of modeling. During the experiment, the mental state, water intake, stool characteristics and the presence of blood in the stool of the mice were closely monitored every day, and the changes in the weight of the mice were recorded.

[0047] 2.5 Sample collection

[0048] Mouse feces were collected at fixed times during the experiment, and fecal occult blood tests were performed according to the instructions. On the 7th day, the mice were placed in metabolic cages, and fecal samples were collected on ice for 4 hours, which were quickly placed in a -80°C refrigerator for testing. On the 8th day, the mice were anesthetized and killed, and the colon was removed. The length of the colon was measured and recorded, the colon was unfolded along the mesentery, and the contents of the colon were collected. The colon was washed with pre-cooled saline, and the colon of 0.5-1 cm at the distal end was cut off and fixed in 4% paraformaldehyde. The remaining colon was divided into 4 parts and quickly frozen and stored in a -80°C refrigerator for later use.

[0049] 2.6 Disease Activity Index in Mice

[0050] Each group of mice was scored according to the mouse disease activity index (DAI) scoring standard (Table 1), and the DAI value of each group of mice was calculated as follows:

[0051] DAI = weight loss percentage score + stool form score + stool blood score

[0052] Table 1 DAI scoring criteria

[0053]

[0054] Note: The fecal occult blood test was performed according to the kit instructions.

[0055] 2.7 Pathological sections

[0056] Take the colon tissue fixed in 4% paraformaldehyde under "2.5", dehydrate, embed, and slice; dewax with xylene to water, stain with hematoxylin and eosin, dehydrate with gradient alcohol, make xylene transparent, and seal with neutral gum. Use a digital slice scanner to collect images of the slices. First, observe the gross lesions of all tissues at low magnification for each slice, and select the area to be observed to collect 100x images to observe specific lesions.

[0057] 2.8 Enzyme-linked immunosorbent assay

[0058] An appropriate amount of colon stored in a -80°C refrigerator was taken and thawed on ice, and the levels of TNF-α, IL-6 and IL-1β in the colon were determined according to the instructions of the ELISA kit.

[0059] 2.916SrDNA high-throughput sequencing experimental process

[0060] The colon contents collected from the normal group, model group, medium-dose group of Litsea cubeba oil and Litsea cubeba water extract were sequenced for 16S rDNA high-throughput sequencing. The experimental process was as follows Figure 2 The primer information is shown in Table 2.

[0061] Table 2 Primer sequence information

[0062]

[0063] 2.10 GC method for detecting the content of short-chain fatty acids in feces

[0064] Short-chain fatty acids are the main metabolites of intestinal bacteria, including acetic acid, propionic acid and butyric acid, and play an important physiological function in the intestine. Short-chain fatty acids can provide energy for intestinal epithelial cells, protect the integrity of the intestinal mucosal barrier, and help improve IBD. Therefore, it is necessary to detect the levels of short-chain fatty acids in each group of mice.

[0065] 2.10.1 Preparation of Standards

[0066] Accurately weigh appropriate amounts of acetic acid, propionic acid, n-butyric acid, isobutyric acid, n-valeric acid, and isovaleric acid reference substances, place them in 10 mL brown volumetric bottles, add ether to dissolve and dilute to the mark. Obtain the reference substance stock solutions of each component, with concentrations of acetic acid (1.350 mg / mL), propionic acid (1.515 mg / mL), butyric acid (1.140 mg / mL), isobutyric acid (0.898 mg / mL), valeric acid (0.398 mg / mL), and isovaleric acid (0.381 mg / mL), respectively, and store at -20°C for later use. Accurately pipette appropriate amounts of each reference substance solution to prepare a series of mixed reference substance solutions.

[0067] 2.10.2 Sample processing method

[0068] The feces collected from the normal group, model group, medium-dose group of Litsea cubeba oil and Litsea cubeba water extract were used for the determination of short-chain fatty acids. 150 mg of feces was weighed into a 1.5 mL EP tube, 500 μL of ultrapure water was added, vortexed for 2 min, and centrifuged at 10000 rpm at 4 ° C for 10 min. 200 μL of the supernatant was taken, 3 μL of 50% sulfuric acid and 400 μL of ether were added, vortexed for 2 min, and centrifuged at 10000 rpm at 4 ° C for 10 min. After standing at 4 ° C for 30 min, the ether layer was sampled and analyzed.

[0069] 2.10.3 Establishment of GC method

[0070] Chromatographic column: Intercap WAX capillary column (0.32 mm × 30 m, 0.50 μm); injection mode: splitless; pressure: 80 kPa; injection volume: 3 μL; injection port temperature: 250 ° C; carrier gas: high purity nitrogen (purity> 99.999%); detector (FID) temperature: 250 ° C; the program temperature is detailed in Table 3.

[0071] Table 3 Program temperature conditions

[0072]

[0073] 3. Results

[0074] 3.1 Effects of Litsea cubeba oil and Litsea cubeba water extract on the physical signs of mice

[0075] like Figure 3As shown in A, the weight of mice in the normal group remained basically unchanged, and the weight of the model group and other intervention groups did not change much in the first two days, but decreased to varying degrees on the third day, while sulfasalazine, Litsea cubeba oil and Litsea cubeba water extract could alleviate the weight loss. During the experiment, the feces of mice in the normal group were normally formed and there was no blood in the stool; mice in the model group began to have mild diarrhea after 3 days, and the severity gradually increased as the experiment progressed, showing watery feces and macroscopic bloody stools; the fecal status of mice in the sulfasalazine, Litsea cubeba oil and Litsea cubeba water extract groups was significantly improved, and blood in the stool and loose stools were reduced.

[0076] 3.2 Effects of Litsea cubeba oil and Litsea cubeba aqueous extract on colon length

[0077] Results Figure 4 Compared with the normal group, the colon length of the mice in the model group was significantly shortened (P<0.001); compared with the model group, intervention with sulfasalazine, Litsea cubeba oil and Litsea cubeba water extract could significantly improve the shortening of colon length (P<0.01).

[0078] 3.3 Effects of Litsea cubeba oil and Litsea cubeba water extract on colonic histopathological morphology

[0079] Pathological examination results showed that ( Figure 5 ), the colon tissue structure of mice in the normal group was intact, the intestinal epithelial cells were arranged intact, and there was no obvious inflammatory cell infiltration and fibrosis; compared with the normal group, the colon tissue of the model group showed a large amount of mucosal layer degeneration and necrosis, intestinal epithelial cells were missing, inflammatory cells infiltrated, and fibrosis was hyperplasia, indicating that the model was successfully established. Sulfasalazine, Litsea cubeba oil and Litsea cubeba water extract can reverse the pathological damage of colon tissue in IBD mice after intervention.

[0080] 3.4 Effects of Litsea cubeba oil and Litsea cubeba water extract on inflammatory factors in the colon

[0081] like Figures 6 to 8 As shown in the figure, compared with the normal group, the levels of TNF-α, IL-6 and IL-1β in the colon of the model group mice were significantly increased (P<0.001), indicating that the model was successfully established. Compared with the model group, after intervention with sulfasalazine, Litsea cubeba oil and water extract, the levels of TNF-α, IL-6 and IL-1β were significantly reduced (P<0.05) in a dose-dependent manner, indicating that sulfasalazine, Litsea cubeba oil and water extract can significantly reverse the changes in inflammatory response in IBD model mice and have an inhibitory effect on the inflammatory response in IBD model mice.

[0082] 3.5 Effects of Litsea cubeba oil and Litsea cubeba aqueous extract on the composition and abundance of intestinal flora in mice 3.5.1 Alpha diversity analysis

[0083] Alpha diversity reflects the species abundance and species diversity of a single sample. There are many measurement indicators: the Sob index and ACE index reflect the species richness of the sample, and the Shannon index and Simpson index comprehensively reflect the species richness and uniformity. Fig. 9 The results showed that compared with the normal group, the α diversity (ACE, Chao1, Shannon and Simpson) of the intestinal flora of mice was not significantly reduced after dextran sulfate sodium induction. There was no significant difference in α diversity between the intervention group of Litsea cubeba oil and Litsea cubeba water extract and the model group.

[0084] 3.5.2β diversity analysis

[0085] β diversity analysis can compare the similarities between different groups in terms of species diversity. Fig.10 As shown in the figure, PcoA analysis showed that compared with the normal group, the composition of the model group's flora was significantly different from that of the normal group, indicating that the flora structure was changed after induction with sodium dextran sulfate. After intervention with Litsea cubeba oil and Litsea cubeba water extract, the flora structure approached that of the normal group. It is speculated that Litsea cubeba oil and Litsea cubeba water extract can change the overall flora structure of IBD model mice and make the community composition of the intestinal flora of mice approach that of normal mice.

[0086] 3.5.3 Species composition analysis

[0087] To further evaluate whether Litsea cubeba oil and Litsea cubeba aqueous extract can affect the composition of the intestinal flora of mice under pathological conditions, we analyzed the relative abundance of the intestinal flora at the phylum, family, and genus levels.

[0088] 3.5.3.1 Species composition of each group at the phylum level

[0089] At the door level, e.g. Fig.11 As shown in the figure, the dominant phyla in each group were Firmicutes, Bacteroidetes, Proteobacteria, and Verrucomicrobiota. Compared with the normal group, the relative abundance ratio of Firmicutes in the fecal microbial community of mice treated with sodium dextran sulfate increased, while the relative abundance ratio of Proteobacteria and Verrucomicrobiota decreased; after intervention with Litsea cubeba oil and water extract, the relative abundance of Firmicutes was significantly reduced, and the relative abundance of Proteobacteria and Verrucomicrobiota was increased, making the relative abundance trend of these flora consistent with that of the normal group. This shows that both Litsea cubeba oil and Litsea cubeba water extract can adjust the intestinal community structure at the phylum level.

[0090] 3.5.3.2 Species composition of each group at the family level

[0091] like Fig.12 As shown in the figure, at the family level, compared with the normal group, the microbiota of the model group changed significantly, increasing the relative abundance of Lachnospiraceae, Prevotellaceae, Erysipelotrichaceae, and Oscillospiraceae, and decreasing the relative abundance of Enterobacteriaceae, Muribaculaceae, Bacteroidaceae, Akkermansiaceae, Tannerellaceae, and Lactobacillaceae. After intervention with Litsea cubeba oil and Litsea cubeba water extract, both of them could restore the Enterobacteriaceae, Lachnospiraceae, Bacteroidaceae, Akkermansiaceae, Erysipelotrichaceae, Prevotellaceae and Tannerellaceae, making the relative abundance trend of these bacterial communities consistent with that of the normal control group, suggesting that Litsea cubeba oil and Litsea cubeba water extract have the function of adjusting the intestinal community structure at the family level.

[0092] 3.5.3.3 Species composition of each group at the genus level

[0093] like Fig.13 As shown in the figure, at the genus level, compared with the normal group, the microbiota of the model group changed significantly, increasing the relative abundance of Eubacterium fissicatena group, Prevotellaceae UCG-001, Dubosiella, and Lachnospiraceae NK4A136 group, and decreasing the relative abundance of Escherichia-Shigella, Bacteroides, Enterobacter, Akkermansia, Lactobacillus, and Parabacteroides.

[0094] After intervention with Litsea cubeba oil and Litsea cubeba water extract, Escherichia-Shigella, Enterobacter, Akkermansia, Prevotellaceae_UCG-001, Parabacteroides, Dubosiella, and the water extract could also adjust Eubacterium_fissicatena_group, making the relative abundance trends of these bacterial communities consistent with that of the normal control group, indicating that Litsea cubeba oil and water extract also have the function of adjusting the intestinal community structure at the genus level.

[0095] 3.6 Short-chain fatty acid content determination results

[0096] The contents of 6 short-chain fatty acids in the feces of mice in each group were determined. Fig.14 ): Compared with the normal group, the fecal contents of acetic acid, propionic acid, butyric acid and valeric acid in the model group were significantly decreased (P<0.05). After intervention with Litsea cubeba oil and Litsea cubeba water extract, the contents of acetic acid and butyric acid were significantly increased (P<0.05), suggesting that both can regulate short-chain fatty acids.

[0097] 4. Summary

[0098] Litsea cubeba oil and Litsea cubeba water extract can alleviate the weight loss of IBD mice, reduce the DAI score, significantly restore the colon length (P<0.05), reduce the pathological damage of colon tissue, and significantly reduce the content of proinflammatory cytokines TNF-α, IL-6 and IL-1β in the colon (P<0.05). The results of α diversity and β diversity analysis showed that Litsea cubeba oil and water extract can change the overall microbiota structure of DSS-induced IBD mice, making the community composition of the intestinal flora of mice closer to that of the normal group. Further analysis of the relative abundance of intestinal flora at the phylum, family and genus levels showed that Litsea cubeba oil and Litsea cubeba water extract can affect the composition of the intestinal flora of mice under pathological conditions. By measuring the contents of 6 short-chain fatty acids in the feces of each group of mice, it was found that the contents of acetic acid and butyric acid were significantly increased after intervention with Litsea cubeba oil and water extract (P<0.05), suggesting that Litsea cubeba oil and Litsea cubeba water extract have preventive and therapeutic effects on inflammatory bowel disease.

[0099] Finally, it should be pointed out that the above embodiments are only representative examples of the present invention. Obviously, the technical solution of the present invention is not limited to the above embodiments, and there are many variations. All variations that can be directly derived or associated with the content disclosed by ordinary technicians in this field should be considered as the protection scope of the present invention.

Claims

1. Application of Litsea cubeba in the preparation of drugs for inflammatory bowel disease.

2. Use of the Litsea cubeba as claimed in claim 1 in preparing a drug for inflammatory bowel disease, It is characterized in that The macrocarpon Litsea cubeba includes products made from the macrocarpon Litsea cubeba.

3. Use of the Litsea cubeba as claimed in claim 2 in the preparation of inflammatory bowel disease drugs, It is characterized in that The Litsea cubeba described is Litsea cubeba oil.

4. Use of the Litsea cubeba as claimed in claim 2 in preparing a drug for inflammatory bowel disease, It is characterized in that The Litsea cubeba is a water extract of Litsea cubeba.

5. Use of the Litsea cubeba as claimed in claim 1 in preparing a drug for inflammatory bowel disease, It is characterized in that The inflammatory bowel disease drug is a drug for preventing or treating inflammatory bowel disease.

6. Use of the Litsea cubeba as claimed in claim 1 in preparing a drug for inflammatory bowel disease, It is characterized in that The inflammatory bowel disease drug is a drug for reducing the symptoms of bloody stools and loose stools caused by inflammatory bowel disease.

7. Use of the Litsea cubeba as claimed in claim 1 in preparing a drug for inflammatory bowel disease, It is characterized in that The inflammatory bowel disease drug is a drug that improves the symptoms of shortened colon length.

8. Use of the Litsea cubeba as claimed in claim 1 in preparing a drug for inflammatory bowel disease, It is characterized in that The inflammatory bowel disease drug is a drug that reverses pathological damage to colon tissue.

9. Use of the Litsea cubeba as claimed in claim 1 in preparing a drug for inflammatory bowel disease, It is characterized in that The inflammatory bowel disease drug is a drug that reverses changes in inflammatory responses, inhibits inflammatory responses, and reduces the levels of TNF-α, IL-6, and IL-1β.

10. Use of the Litsea cubeba as claimed in claim 1 in preparing a drug for inflammatory bowel disease, It is characterized in that The inflammatory bowel disease drug is a drug that adjusts the intestinal flora structure toward normal.

Images