Effect of cordycepin in field of fat protection

Abstract

An effect of cordycepin in the field of fat protection, specifically an application of cordycepin in the preparation of fat protection products in the field of treatment of mental illness or sub-healthy mental state, the therapeutic dose is 25 mg cordycepin per kilogram of body weight. 25 mg / kg cordycepin can increase the weight reduction of white adipose tissue (sWAT) and perirenal white adipose tissue (pWAT) induced by stress, significantly reduce the abundance of Prevotellaceae UCG-001, increase the abundance of Desulfovibrio, and have an anti-anxiety or depression tendency.

Description

The role of cordycepin in fat protection Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to the effect of cordycepin in the field of fat protection. Background Art

[0002] Anxiety disorders (including generalized anxiety disorder and panic disorder) are among the most prevalent and difficult-to-treat psychiatric illnesses. Anxiety and depression are widespread and debilitating mental illnesses, affecting approximately 10% of the global population annually (World Health Organization, 2017). Although the use of psychotherapy and psychotropic medications has increased over the past few decades, the results are often suboptimal, and the prevalence of anxiety and depressive disorders remains unchanged. Poria cocos (P. cocos) has been used as a folk medicine and functional food in China for over 2,000 years. Chronic sleep deprivation (CSD) induces anxiety, intestinal dysbiosis, and inflammation, and reduces neurotransmitter levels. P. cocos treatment improves anxiety-like behaviors in CSD rats, significantly increases the species richness and diversity of the gut microbiota in anxious rats, and regulates the abundance of the dysregulated Prevotellaceae_UCG-001 to levels close to those in normal controls. Neuroinflammation is a central factor in neuropathic pain (NP). Ginger, with its anti-inflammatory properties, is a promising bioactive compound for the treatment of NP. Emerging evidence suggests that the gut microbiome and gut-derived metabolites play a key role in NP. Two ginger root extracts, rich in gingerol (GEG) and shogaol (SEG), have a certain impact on pain sensitivity, anxiety-like behavior, gut microbial composition, and fecal metabolites in rats with NP. Compared with the spinal nerve ligation (SNL) group, the SNL+GEG and SNL+SEG groups showed significant reductions in pain and anxiety-like behavior, as well as reduced abundance of Prevotellaceae UCG-001 and Desulfovibrio.

[0003] Diabetes is a major threat to human health. The "Global Distribution of Diabetes," published on the International Diabetes Federation (IDF) website, shows that in 2019, 463 million people worldwide suffered from diabetes, a number projected to reach 700 million by 2045. Type 2 diabetes mellitus (T2DM) is the most common type of diabetes, accounting for over 90% of all diabetes diagnoses. Dyslipidemia, gut microbiome imbalance, and abnormal liver lipid metabolism are key features of T2DM. Supplementation with Jerusalem artichoke inulin can improve abnormal liver lipid metabolism by altering the expression of genes involved in lipid and cholesterol production and degradation. Microbiological analysis has shown that supplementation with Jerusalem artichoke inulin can enrich the Prevotellaceae UCG-001. Studies have shown that anthocyanins are the most abundant bioactive compounds in black corn, a pigmented corn variety. 16S ribosomal RNA sequencing analysis of cecal microbiota revealed that the consumption of black corn flour increased the relative abundance of Prevotellaceae UCG-001. Obesity is a multifactorial disease associated with the development of various chronic diseases. Growing evidence suggests that obesity and obesity-related metabolic diseases are significantly influenced by the gut microbiome. Fuzhuan tea, a traditional Chinese tea, has long been used as a traditional treatment for hypercholesterolemia, hyperglycemia, metabolic disorders, and gastrointestinal disorders. Adding Fuzhuan tea to dietary polysaccharides can significantly reduce the increase in Desulfovibrio abundance caused by dietary polysaccharides.

[0004] Cordycepin (3′-deoxyadenosine, Cpn) is the main bioactive component of Cordyceps militaris. It has a wide range of biological activities, such as anti-inflammatory, antioxidant, anti-fibrotic, anti-adipogenic and anti-tumor activities, and is currently the focus of research.

[0005] Summary of the Invention

[0006] In order to solve the above technical problems, the present invention provides an intestinal microorganism that mediates the fat-protecting effect of cordycepin.

[0007] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0008] The present invention provides a role of cordycepin in the field of fat protection. The role of cordycepin in the field of fat protection is specifically: use in the preparation of fat-protecting products for treating mental illnesses / sub-healthy mental states, with a therapeutic dose of 25 mg of cordycepin per kilogram of body weight.

[0009] Furthermore, the therapeutic dose is used to reduce the abundance of Prevotellaceae UCG-001 and increase the abundance of Desulfovibrio.

[0010] Furthermore, the product is a pharmaceutical composition or a food.

[0011] Furthermore, the dosage form of the pharmaceutical composition is any one of injection, tablet, capsule, suspension, ointment, microsphere, aerosol, inhalant and pill.

[0012] Furthermore, the pharmaceutical composition further comprises any one or a combination of pharmaceutically acceptable excipients, carriers and matrices.

[0013] Furthermore, the food types include: health food and beverages.

[0014] Furthermore, the mental illness includes anxiety, depression, and Alzheimer's disease.

[0015] Furthermore, the sub-healthy mental state includes distraction, fatigue, anxiety, and insomnia.

[0016] Compared with the existing technology, the technical solution provided by the present invention has at least the following advantages:

[0017] The present invention provides an intestinal microbe that mediates the protective effect of cordycepin on fat. This study is the first to discuss the effects of different doses of cordycepin on anti-anxiety / depression, the changes in intestinal microbial richness, and the effects on fat mass. The present invention has undergone experiments and simulations to prove the feasibility of the experiment. The results showed that 25mg / kg cordycepin can increase the weight loss of white adipose tissue (sWAT) and perirenal white adipose tissue (pWAT) induced by stress, significantly reduce the abundance of Prevotellaceae UCG-001, and increase the abundance of Desulfovibrio, showing a trend of anti-anxiety / depression. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] One or more embodiments are exemplarily described by the pictures in the corresponding drawings. These exemplifications do not constitute limitations on the embodiments. Unless otherwise stated, the pictures in the drawings do not constitute proportional limitations.

[0019] FIG1 is a diagram showing how cordycepin improves anxiety / depression behavior in mice according to an embodiment of the present invention;

[0020] FIG2 is a graph showing the adipose tissue weights of mice in different groups according to an embodiment of the present invention;

[0021] FIG3 is a diagram showing the regulation of the richness of intestinal microbiota mediated by cordycepin-induced stress damage in an embodiment of the present invention. DETAILED DESCRIPTION

[0022] The inventors found that no existing studies have linked anti-anxiety / depression with gut microbes and fat weight.

[0023] In peripheral tissues, Cpn plays an important protective and regulatory role in diabetes and adipose tissue abnormalities. Takahashi reported that Cpn exerts anti-adipogenic effects by intervening in the activation of the C / ebpb-ppar pathway. Later studies found that high concentrations of Cpn could significantly reduce body weight, visceral fat index, total cholesterol (TC) levels, triglyceride (TG) levels, low-density lipoprotein (LDL) levels, and high-density lipoprotein (HDL) levels in mice fed a high-fat diet; although low concentrations of Cpn still reduced body weight, visceral fat index, serum TG and LDL levels, it had no effect on serum TC and HDL levels. Another study showed that Cpn has an antidepressant effect in the tail suspension test. In addition, TST and FST observations showed that Cpn had a beneficial effect on CMS-induced behavioral deficits after 3-6 weeks of administration at doses of 20 mg / kg and 40 mg / kg. The study also found that Cpn could improve metabolic disorders by regulating the homeostasis of gut microbiota (GM), including selectively increasing the abundance of Akkermansia muciniphila.

[0024] The present invention aims to explore the effects of different doses of cordycepin on anti-anxiety and depression, intestinal microorganisms and fat weight.

[0025] The present invention provides a role of cordycepin in the field of fat protection. The role of cordycepin in the field of fat protection is specifically: use in the preparation of fat-protecting products for treating mental illnesses / sub-healthy mental states, with a therapeutic dose of 25 mg of cordycepin per kilogram of body weight.

[0026] Furthermore, the therapeutic dose is used to reduce the abundance of Prevotellaceae UCG-001 and increase the abundance of Desulfovibrio.

[0027] Furthermore, the product is a pharmaceutical composition or a food.

[0028] Furthermore, the dosage form of the pharmaceutical composition is any one of injection, tablet, capsule, suspension, ointment, microsphere, aerosol, inhalant and pill.

[0029] Furthermore, the pharmaceutical composition further comprises any one or a combination of pharmaceutically acceptable excipients, carriers and matrices.

[0030] Furthermore, the food types include: health food and beverages.

[0031] Furthermore, the mental illness includes anxiety, depression, and Alzheimer's disease.

[0032] Furthermore, the sub-healthy mental state includes distraction, fatigue, anxiety, and insomnia.

[0033] Specifically, in the present invention, the specific testing method is:

[0034] Step 1: In the present invention, mice are first gavaged with cordycepin for 4 weeks.

[0035] Step 2: After 4 weeks, stress induction was performed for 4 consecutive days.

[0036] Step 3: After stress, perform open field, elevated plus maze, black-white box, Y maze, and tail suspension tests, collect fecal samples, and weigh the fat of the mice.

[0037] The following describes preferred embodiments of the present invention to make its technical content clearer and easier to understand. The present invention can be embodied in many different forms of embodiments, and the scope of protection of the present invention is not limited to the embodiments mentioned herein.

[0038] Many terms are used in the present invention. Unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. In order to more clearly and consistently understand the present invention, the definitions and explanations of the relevant terms are provided below.

[0039] As used herein, the terms "gavage" and "gavage administration" mean that before administration, the experimental animals are generally fasted for 12 to 16 hours, after which the animals are fixed and the drug solution is injected directly into the animal's stomach through the animal's mouth using a syringe and gavage needle.

[0040] As used in this article, the term "open field test" or "open field test" is also called the open box test, which is a method to evaluate the autonomous behavior, exploratory behavior and tension of experimental animals in a novel environment. The frequency and duration of certain behaviors of experimental animals in a novel environment reflect the autonomous behavior and exploratory behavior of experimental animals in an unfamiliar environment. The open field test observes and studies the neuropsychiatric changes of experimental animals and various behaviors after entering an open environment. For example, the fear of the new open environment causes animals to mainly move in the peripheral area and less in the central area, but the exploratory characteristics of the animals prompt them to generate motivation to move in the central area, and the anxiety caused by this can also be observed. Central nervous system stimulants can significantly increase autonomous activity and reduce exploratory behavior. A certain dose of antipsychotic drugs can reduce exploratory behavior without affecting autonomous activity.

[0041] As used herein, the terms "elevated plus maze test" and "elevated plus maze" refer to the use of conflicting behaviors formed by animals' exploratory characteristics of novel environments and their fear of high-hanging open arms to examine the anxiety state of animals. The elevated plus maze has a pair of open arms and a pair of closed arms. Rodents tend to move in the closed arms due to their dark preferences, but they will move in the open arms out of curiosity and exploratory nature. When faced with novel stimuli, animals simultaneously have the urge to explore and fear, which results in conflicting behaviors of exploration and avoidance, thereby generating anxiety. Anti-anxiety drugs can significantly increase the number and time of entering the open arms. The elevated plus maze is widely used in scientific research and computer-assisted teaching in multiple disciplines such as new drug development / screening / evaluation, pharmacology, toxicology, preventive medicine, neurobiology, animal psychology and behavioral biology. It is a classic experiment for conducting behavioral research, especially anxiety and depression research.

[0042] As used herein, the term "light and dark box" refers to a light-dark box experiment in which mice or rats prefer to explore the dark box (black box), but their exploratory nature prompts them to attempt to explore the light box (white box). However, the bright light stimulation of the light box inhibits the animals' exploratory activity in the light box. Anxiolysis can relieve this inhibitory effect.

[0043] As used herein, the term "Y-maze behavioral experiment" is a symmetrical, closed, three-arm maze, shaped like a capital "Y," with an angle of 120 ° between each arm, and each arm having a removable gate for limiting the activity of the experimental animals. The three arms are randomly set to: a novel arm, a start arm, and the other arm. The Y-maze arm length for rats is 50 cm, the maze passage is approximately 10 cm wide, and the maze wall is approximately 13 cm high; the Y-maze arm length for mice is 28 cm, the maze passage is approximately 5 cm wide, and the maze wall is approximately 10 cm high. The entire maze is approximately 50 cm above the ground, and the maze is typically painted dark to prevent animals from experiencing anxiety during the experiment. The Y-maze can be used to assess the short-term memory of mice and rats, and also allows mice to assess spontaneous alternation by exploring all three arms of the maze, which is a measure of spatial working memory and is driven by the innate curiosity of rodents to explore areas that have not been visited before. Mice with intact working memory and intact prefrontal cortex function remember previously visited maze arms and show a tendency to enter more recently visited maze arms. Spatial reference memory, which is emphasized by the hippocampus, can also be tested by placing test mice in a Y-maze with one arm closed during training.

[0044] As used herein, the "tail suspension test" is a classic and rapid method for evaluating the efficacy of antidepressants, stimulants, and sedatives. It utilizes the principle that mice, after being suspended by their tails, attempt to escape but are unable to do so, resulting in them giving up their struggle and entering a unique state of depressive immobility. During the experiment, the duration of immobility is recorded to reflect the animal's depressive state. Antidepressants and stimulants can significantly shorten or alter this state.

[0045] The present invention will be further described below through specific examples.

[0046] The experimental mice were subjected to a 4-day repeated stress protocol consisting of the following steps: (i) restraint for 1 hour, (ii) tail suspension for 30 minutes, and (iii) foot shock for 10 cycles (0.5 mA, 2 seconds, 10 seconds interval). The groups were as follows:

[0047] Control + water: no stress + pure water gavage;

[0048] Stress + water: stress + pure water gavage;

[0049] Stress+12.5Cpn: stress+12.5mg / kg cordycepin gavage;

[0050] Stress+25Cpn: stress+25mg / kg cordycepin gavage;

[0051] No stress + 25Cpn: No stress + 25 mg / kg cordycepin administered orally.

[0052] After four weeks of oral administration, mice were tested in the open field (OFT), elevated plus maze (EPM), black-and-white box (LDB), Y-maze (Y-maze), and tail suspension test (TST). Feces were collected for 16S ribosomal RNA sequencing, and subcutaneous white adipose tissue (sWAT), perirenal white adipose tissue (pWAT), epididymal white adipose tissue (eWAT), and brown adipose tissue (BAT) were measured.

[0053] Figure 1 shows the improvement of anxiety / depression behavior in mice using cordycepin according to an embodiment of the present invention. In Figure 1, (A) shows the movement trajectory, intermediate dwell time, total distance traveled, intermediate distance / total distance traveled, and number of intermediate entries in the OFT test; (B) shows the movement trajectory, open arm dwell time, total immobility time, number of open arm entries, and open arm distance traveled in the EPM test; (C) shows the latency, white box dwell time, and number of white / black box crossings in the LDB test; (D) shows the number of spontaneous alternations and total number of arm entries in the Y-maze test; and (E) shows the immobility time in the TST. P values ​​were calculated using an unpaired t-test or one-way ANOVA; the remaining groups were analyzed using Tukey's multiple comparison test. Bar graphs represent mean ± SEM values. Behavior: N = 9-10 / group; CORT: N = 5 / group. Control + Water Group: No Stress + Pure Water Gavage; Stress + Water Group: Stress + Pure Water Gavage; Stress + 12.5 Cpn: Stress + 12.5 mg / kg Cordycepin Gavage; Stress + 25 Cpn: Stress + 25 mg / kg Cordycepin Gavage; No Stress + 25 Cpn: No Stress + 25 mg / kg Cordycepin Gavage. OFT: Open Field Test; EPM: Elevated Plus Maze; LDB: Black-White Box; Y-maze: Y-maze; TST: Tail Suspension Test.

[0054] Figure 2 is a graph of adipose tissue weights of mice in different groups according to the present invention. #p<0.05, **p<0.01, ##p<0.01, determined by unpaired t-test or one-way ANOVA with Tukey's multiple comparison test. Bar graphs represent mean ± SEM values. N = 5 per group. Control + water group: no stress + pure water gavage; Stress + water group: stress + pure water gavage; Stress + 12.5 Cpn group: stress + 12.5 mg / kg cordycepin gavage; Stress + 25 Cpn group: stress + 25 mg / kg cordycepin gavage; No stress + 25 Cpn group: no stress + 25 mg / kg cordycepin gavage. pWAT: perirenal white adipose tissue; eWAT: epididymal white adipose tissue; sWAT: subcutaneous white adipose tissue; BAT: brown adipose tissue.

[0055] Figure 3 is a graph showing the regulation of stress-induced damage to intestinal microbiota richness mediated by cordycepin in an embodiment of the present invention. In Figure 3, (A) shows the relative abundance of Prevotellaceae_UCG-001, (B) shows the relative abundance of NK4A214_group, and (C) shows the relative abundance of Desulfovibrio. #p<0.05, *P<0.05, **P<0.01, determined by unpaired t-test or one-way ANOVA with Tukey's multiple comparison test. Bar graph represents mean ± SEM. N = 4-5 per group. Control + water group: no stress + pure water gavage; Stress + water group: stress + pure water gavage; Stress + 12.5 Cpn group: stress + 12.5 mg / kg cordycepin gavage; Stress + 25 Cpn group: stress + 25 mg / kg cordycepin gavage; No stress + 25 Cpn group: no stress + 25 mg / kg cordycepin gavage.

[0056] In summary, although there was no significant difference in the effect of 25 mg / kg cordycepin compared to the stress group, it showed an anti-anxiety / depression trend. 25 mg / kg cordycepin significantly increased the weight loss of white adipose tissue (sWAT) and perirenal white adipose tissue (pWAT) induced by stress, but had no effect on the weight of epididymal white adipose tissue (eWAT) and brown adipose tissue (BAT).

[0057] The present invention has been tested and simulated to demonstrate experimental feasibility. The results show that 25 mg / kg of cordycepin can increase stress-induced weight loss in white adipose tissue (sWAT) and perirenal white adipose tissue (pWAT), significantly reduce the abundance of Prevotellaceae UCG-001, and increase the abundance of Desulfovibrio, showing an anti-anxiety / depression effect.

[0058] Those skilled in the art will appreciate that the above-described embodiments are specific examples for implementing the present application, and that in actual applications, various changes in form and detail may be made thereto without departing from the spirit and scope of the present application. Any person skilled in the art may make changes and modifications without departing from the spirit and scope of the present application. Therefore, the scope of protection of the present application shall be subject to the scope defined in the claims.

Claims

1. The role of cordycepin in the field of fat protection is characterized by: The role of cordycepin in the field of fat protection is specifically: its application in the preparation of fat protection products for the treatment of mental illnesses / sub-healthy mental states, with a therapeutic dose of 25 mg of cordycepin per kilogram of body weight.

2. The use according to claim 1, characterized in that: The therapeutic dose was used to reduce the abundance of Prevotellaceae UCG-001 and increase the abundance of Desulfovibrio.

3. The use according to claim 1, characterized in that: The product is a pharmaceutical composition or a food.

4. The use according to claim 3, characterized in that: The dosage form of the pharmaceutical composition is any one of injection, tablet, capsule, suspension, ointment, microsphere, aerosol, inhalant and pill.

5. The use according to claim 4, characterized in that: The pharmaceutical composition further comprises any one or a combination of pharmaceutically acceptable excipients, carriers and matrices.

6. The use according to claim 1, characterized in that: The food types include: health food and beverages.

7. The use according to claim 1, characterized in that: The mental illnesses include anxiety, depression, and Alzheimer's disease.

8. The use according to claim 1, characterized in that: The sub-healthy mental state includes distraction, fatigue, anxiety, and insomnia.

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