Application of ganoderic acid A in preparation of medicine for treating irritable bowel syndrome

Abstract

The invention discloses application of ganoderic acid A in treatment of irritable bowel syndrome for the first time. The invention specifically relates to application of ganoderic acid A in improving IBS mouse intestinal total transport time and colon transport speed abnormity, reducing IBS mouse excrement water content and pain response, relieving diarrhea and visceral hypersensitivity symptoms, and improving IBS mouse colon intestinal mucosa structure integrity. The ganoderic acid A provided by the invention can be used for application and research and development of drugs or functional products for preventing and treating irritable bowel syndrome.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to the application of ganoderic acid A in the preparation of drugs and functional products for the prevention and treatment of irritable bowel syndrome. Background Technology

[0002] Anxiety and depression, social stress, low-grade inflammation of the intestinal mucosa, and impaired gut microbiota can lead to intestinal dysfunction and motility disorders, and even induce irritable bowel syndrome (IBS). IBS is a common gut-brain interaction disorder (DGBI), mainly characterized by abdominal pain, bloating, diarrhea, or constipation. Studies have shown that the prevalence of IBS varies across different countries and regions, with a prevalence of 17%–21% in South America, 7%–9% in South Asia, and 5.6% in the Middle East and Africa. The prevalence of IBS in women is 1.5–3 times that in men. The incidence of IBS also differs across age groups, particularly among young and middle-aged adults (aged 18–59 years), where the prevalence is higher. In China, the prevalence of IBS in adults ranges from 2.3% to 15.8%.

[0003] Currently, the exact pathological mechanism of IBS has not been fully elucidated, and there is a lack of effective treatment drugs. Nevertheless, some factors such as visceral hypersensitivity, intestinal inflammatory response, disruption of intestinal barrier function, and imbalance of gut microbiota are believed to play a crucial role in the pathogenesis of IBS. (1) Visceral hypersensitivity: Existing evidence suggests that 30% to 40% of IBS patients have higher sensitivity to colonic dilatation, manifested as a lower pain threshold and increased intensity of sensation to colonic dilatation. Therefore, there is a link between the pathogenesis of IBS and visceral hypersensitivity. Visceral hypersensitivity can serve as a clinical marker of IBS and can explain the abdominal discomfort symptoms such as constipation and diarrhea in IBS patients. (2) Intestinal inflammation: A recent study found that some IBS patients had elevated levels of pro-inflammatory factors such as IL-1β, IL-6, IL-8, IL-12, TNF-α, T cells, B cells, and macrophages in their serum, and the severity of symptoms was correlated with the levels of inflammatory markers. Therefore, inflammation may be a susceptibility factor for IBS. (3) Intestinal barrier damage: The intestinal barrier protects us from threats from the intestinal lumen and plays a vital role in maintaining intestinal homeostasis. Tight junctions and adhesive junctions are important components of the intestinal mucosal barrier. Therefore, developing a natural medicine or functional food with multiple beneficial effects for IBS based on the above factors will provide a new theoretical basis for the treatment of intestinal diseases with natural products.

[0004] Reishi mushroom, a traditional Chinese medicine, has a long history and is known as the "King of Herbs." In recent years, it has been included in the "List of Food and Medicine with the Same Origin," highlighting its various beneficial effects on human health. It is believed to have effects such as lowering blood sugar, clearing heat and detoxifying, and anti-tumor properties, and is widely used in traditional Chinese medicine and naturopathy, highly regarded by doctors and naturopaths. In recent years, with a deeper understanding of traditional Chinese medicine, the medicinal value of reishi mushroom has been continuously discovered and recognized. Ganoderic acid A is an active ingredient extracted from reishi mushroom. As a natural drug component, it has low toxicity and high safety, and is considered one of its main pharmacological components, possessing potential therapeutic effects. Its content is used as an important indicator for evaluating the quality of reishi mushroom. Ganoderic acid A was first isolated by Kubota et al. in 1982. Belonging to the triterpenoid class of compounds, it currently possesses various pharmacological effects and medical value, including anti-cancer, anti-inflammatory, immunomodulatory, hepatoprotective, antioxidant, and lipid-regulating effects. However, a systematic analysis of the mechanism of action of ganoderic acid A is lacking.

[15] This invention aims to investigate the potential therapeutic effects of ganoderic acid A on IBS, providing new ideas and methods for the treatment of this disease and promoting the development of traditional Chinese medicine. Summary of the Invention

[0005] This invention discloses a novel use of ganoderic acid A (GAA), a lanosterane-type triterpenoid compound derived from the medicinal and edible mushroom Ganoderma lucidum, in the pharmaceutical and food fields. Specifically, ganoderic acid A is used in the preparation of drugs and / or functional products for the prevention and / or treatment of irritable bowel syndrome.

[0006] The structural formula of this compound is:

[0007]

[0008] The dosage forms of drugs include capsules, traditional Chinese medicine decoctions, granules, tablets, ointments, oral liquids, injections, suppositories, or aerosols.

[0009] Functional products include beverages, syrups, biscuits, pastries, teas, or powders.

[0010] Ganoderic acid A is used in the preparation of drugs and / or functional products for the prevention and / or treatment of intestinal motility disorders.

[0011] Ganoderic acid A is used in the preparation of drugs for the prevention and / or treatment of diarrhea and / or functional products.

[0012] Ganoderic acid A is used in the preparation of drugs and / or functional products for the prevention and / or treatment of visceral hypersensitivity.

[0013] The compound ganoderic acid A of this invention, as an inhibitor, has the potential for development and application in the pharmaceutical and food fields. Attached Figure Description

[0014] The accompanying drawings are provided to further illustrate the present disclosure and form part of the specification. They are used together with the following detailed description to explain the present disclosure, but do not constitute a limitation thereof. In the drawings:

[0015] Figure 1 Effect of ganoderic acid A on total intestinal transit time in IBS mice;

[0016] Figure 2 Effects of ganoderic acid A on colonic transport function in IBS mice;

[0017] Figure 3 Effect of ganoderic acid A on fecal water content in IBS mice;

[0018] Figure 4-6 Effects of ganoderic acid A on visceral sensitivity in IBS mice; Figure 4 The score of the abdominal withdrawal reflex in mice under 0.1 ml gas pressure; Figure 5 The score of the abdominal withdrawal reflex in mice under 0.2 ml gas pressure; Figure 6 The score of the abdominal withdrawal reflex in mice under 0.3 ml gas pressure;

[0019] Figure 7 Effects of ganoderic acid A on the pathological damage of colon tissue in IBS mice. Detailed Implementation

[0020] The technical solution of the present invention will be described in detail below with reference to the accompanying drawings:

[0021] A lanosterane-type triterpenoid compound, ganoderic acid A, derived from the medicinal and edible mushroom Ganoderma lucidum, is used in pharmaceuticals and food for the prevention and treatment of irritable bowel syndrome (IBS) in drugs or functional products. The structural formula of this compound is as follows:

[0022]

[0023] In the embodiments of this disclosure, ganoderic acid A can be used to prepare an application that improves the total intestinal transit time in IBS mice.

[0024] In the embodiments of this disclosure, ganoderic acid A can be used to prepare an application that improves abnormal colonic transit velocity in IBS mice.

[0025] In the embodiments of this disclosure, the application of ganoderic acid A in the treatment and relief of intestinal motility disorders was discovered for the first time, and it was found to be superior to IBS-positive drugs.

[0026] In the embodiments disclosed herein, ganoderic acid A can significantly reduce the water content of feces in IBS mice and improve diarrhea symptoms.

[0027] In the embodiments of this disclosure, ganoderic acid A has the effect of relieving visceral hypersensitivity.

[0028] In the embodiments of this disclosure, ganoderic acid A can be used to prepare a product that alleviates histopathological damage to the colon of IBS mice and improves intestinal inflammation.

[0029] In the embodiments of this disclosure, ganoderic acid A can be used in the preparation of drugs and functional products for relieving and treating irritable bowel syndrome.

[0030] In the embodiments of this disclosure, the drug dosage forms mainly include capsules, traditional Chinese medicine decoctions, granules, tablets, ointments, oral liquids, injections, suppositories, and aerosols.

[0031] In embodiments of this disclosure, functional products mainly include beverages, syrups, biscuits, pastries, teas, or powders.

[0032] In the specific implementation of this invention, all experimental steps followed animal regulations and were approved by the animal ethics committee. 1.1 Glossary of terms, the specific name codes are shown in Table 1:

[0033] Table 1 List of abbreviations used

[0034]

[0035] 1.2 Experimental reagents and materials are shown in Table 2:

[0036] Table 2 Commonly Used Main Reagents

[0037]

[0038] 1.3 Experimental instruments and equipment, as detailed in Table 3:

[0039] Table 3 Commonly Used Main Experimental Instruments and Equipment

[0040]

[0041]

[0042] 1.4 Animal Experiment Design

[0043] The specific groupings for the experiment are as follows:

[0044] (A) Blank control group;

[0045] (B) Model Group;

[0046] (C) Positive drug (Pivaverium bromide tablets, Dicetel);

[0047] (D) Ganoderic Acid A (GAA) intervention group;

[0048] SPF-grade male C57BL / 6J mice purchased from Beijing Speford Biotechnology Co., Ltd. were used in the experiment. They were fed standard AIN-93M diet daily with free access to water and housed in the SPF-grade Animal Center of Northwest A&F University. After a 7-day acclimatization period, the experiment officially began. One day prior to the experiment, each mouse was administered 0.2 mL of *Citrobacter muscarinica* suspension by gavage daily, followed by intraperitoneal injection of 0.5 mL of lactated Ringer's solution for 7 consecutive days to prevent dehydration. From day 2 to day 29, mice in the blank control group and IBS model group were administered 0.2 mL of sterile saline by gavage daily, the positive control group was administered pinaverium bromide solution (0.2 mL / mouse / day), and the intervention group was administered 20 mg of ganoderic acid A by gavage daily. From day 17 to day 29, the mice underwent water avoidance stress (WAS). From day 29 to day 31, total intestinal transit time, colonic transit function, fecal water content, and visceral sensitivity were measured before the mice were sacrificed.

[0049] 1.5 Data Statistical Analysis

[0050] All data from this study were analyzed using GraphPad. Differences between groups were compared using one-way ANOVA and Tukey's multiple comparison test. Results are expressed as mean ± standard error. (n=6) indicates. Compared with the control group, # P<0.05 indicates a significant difference. ## P<0.01 indicates a highly significant difference; compared with the IBS group, * P<0.05 indicates a significant difference. ** P<0.01 indicates that the difference is highly significant.

[0051] 1.6 Total transport time of intestines

[0052] Altered intestinal motility is a hallmark of IBS. Using a previous experimental method, carmine dye was prepared into a 6% (w / v) solution in 0.5% methylcellulose. Mice were then gavaged with 200 μL of the 6% (w / v) carmine methylcellulose solution and aliquoted into individual cages for monitoring. Fecal samples were subsequently collected and traced on sterile white paper towels to detect the presence of red carmine dye. The time from gavage administration to the first detection of carmine dye in the feces was recorded as the total intestinal transit time t for that mouse.

[0053] like Figure 1As shown, the intestinal transit time in the IBS model group mice was significantly lower than that in the blank control group, proving that the animal modeling results were consistent with expectations. Both the positive drug group and the ganoderic acid A intervention group significantly increased the total gastrointestinal transit time in IBS mice. Among them, the compound was more effective than the positive drug, indicating that this lanosterane triterpenoid compound may have the application efficacy of improving the total gastrointestinal transit time.

[0054] 1.7 Colonic transport function measurement

[0055] Following previous experimental methods, a bead expulsion test was used to assess distal colonic propulsion in mice. Mice were lightly anesthetized with isoflurane, and 2.0 mm spherical plastic beads coated with glycerin were slowly injected into the distal colon (2 cm) using a syringe. The mice were then placed in individual, unlined cages, and the bead expulsion time t was recorded.

[0056] Through observation and data processing, the study found that, for example Figure 2 As shown, the colonic transit rate in the IBS group was significantly higher than that in the control group, while the positive control group and the drug intervention group improved the abnormal colonic transit function in mice, and ganoderic acid A was superior to the first-line treatment for IBS. These two results indicate that ganoderic acid A has the potential to alleviate intestinal motility abnormalities.

[0057] 1.8 Determination of fecal moisture content

[0058] Intestinal water retention capacity and intestinal fluid secretion status are key indicators for assessing IBS patients. In this experiment, fresh mouse feces were collected, weighed, and then dried overnight at 65°C in an oven. The fecal water content was calculated by weighing the samples again. Figure 3 As shown, the study found that the fecal water content of mice in the IBS group was significantly higher than that in the control group, indicating that the model was successfully established. However, after treatment with Dicetel and ganoderic acid A, the fecal water content of the mice significantly decreased and returned to normal levels. These results indicate that ganoderic acid A has the application of reducing fecal water content and improving diarrhea symptoms in mice.

[0059] 1.9 Visceral Sensitivity Measurement

[0060] Visceral hypersensitivity refers to a weakened tolerance and lowered perception threshold of the body to external stimuli, manifesting as an allergic reaction to pain. This state is a major pathological cause of abdominal pain and discomfort, widely considered the core pathological mechanism and biological characteristic of IBS, and also an important criterion for the successful construction of an IBS model. Using previous laboratory methods, the colorectal distension (CRD) test was employed to measure the abdominal withdrawal reflex (AWR) and the CRD threshold pressure that induces rapid contraction of abdominal muscle tissue in adult mice. After anesthetizing the mice, a medical silicone catheter was lubricated with liquid paraffin and slowly inserted into the colon and rectum of the mice through the anus. The mice were allowed 15–30 minutes to awaken and adapt. During the test, air was injected into the balloon to provide different pressure stimuli, with pressure gradually increasing from 0.1, 0.2, to 0.3 mL, with each pressure expansion lasting 20 seconds. The scoring criteria are shown in Table 4. Each mouse underwent the CRD test every 4 minutes, and the AWR response was recorded, repeated 5 times.

[0061] Table 4 Scoring Criteria

[0062]

[0063]

[0064] Studies have found that, for example Figure 4-6 As shown, the IBS group mice exhibited behaviors such as head shaking, abdominal tightening, and back arching, all of which are manifestations of pain response. These observations indicate that the mice exhibit visceral hypersensitivity. At gas pressures of 0.1 mL, 0.2 mL, and 0.3 mL, ganoderic acid A significantly reduced the pain response in the mice. These results suggest that ganoderic acid A has the potential to alleviate symptoms of visceral hypersensitivity.

[0065] 2.0 Histopathological evaluation of colon tissue

[0066] The collected and processed animal tissues were entrusted to Wuhan Saiweier Biotechnology Co., Ltd. for paraffin embedding, sectioning, and staining. After drying, the sections were observed using an inverted fluorescence microscope. The H&E staining results are as follows: Figure 7 As shown, the colonic mucosa of the control group mice remained intact with no inflammatory cell infiltration, while the colonic lamina propria of the IBS group mice showed significantly increased inflammatory cell infiltration. No obvious pathological changes were observed in the mice treated with ganoderic acid A. This indicates that the compound has a certain alleviating effect on intestinal inflammation.

[0067] The preferred embodiments of this disclosure have been described in detail above with reference to the accompanying drawings. However, this disclosure is not limited to the specific details of the above embodiments. Within the scope of the technical concept of this disclosure, various simple modifications can be made to the technical solutions of this disclosure, and these simple modifications all fall within the protection scope of this disclosure.

[0068] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, this disclosure will not describe the various possible combinations separately.

[0069] Furthermore, various different embodiments of this disclosure can be combined in any way, as long as they do not violate the spirit of this disclosure, they should also be regarded as the content disclosed in this disclosure.

Claims

1. Use of ganoderic acid A for preparing drugs and / or functional products for preventing and / or treating irritable bowel syndrome, wherein the structural formula of ganoderic acid A is:

2. The use according to claim 1, characterized in that: The dosage form of the drug includes capsules, Chinese medicine decoctions, granules, tablets, ointments, oral liquids, injections, suppositories or aerosols.

3. The use according to claim 1, characterized in that: The functional products include beverages, syrups, biscuits, cakes, teas or instant medicines.

4. Application of ganoderic acid A in the preparation of drugs and / or functional products for preventing and / or treating intestinal motility disorders.

5. Use of ganoderic acid A in preparing medicines and / or functional products for preventing and / or treating diarrhea.

6. Use of ganoderic acid A in preparing drugs and / or functional products for preventing and / or treating visceral hypersensitivity.

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