Effect of cordycepin in field of fat protection

Abstract

The invention provides an effect of cordycepin in the field of fat protection, and particularly relates to an application of cordycepin in preparation of products for protecting fat in the field of treating psychotic diseases / sub-health mental states, and the therapeutic dose is that 25mg of cordycepin is given per kilogram of body weight. The experimental feasibility is proved through experiments and simulation. Results prove that 25 mg / kg of cordycepin can increase the weight reduction of white adipose tissues (sWAT) and perirenal white adipose tissues (pWAT) induced by pressure stress, the richness of PrevoteracaeUCG-001 is remarkably reduced, the richness of Desulfovibrio is increased, and the cordycepin has the tendency of resisting anxiety / depression.

Description

Technical Field

[0001] The 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 the most prevalent and difficult to treat psychiatric disorders. Anxiety and depression are widespread and debilitating psychiatric disorders, affecting approximately 10% of the global population each year (World Health Organization, 2017). Although the use of psychotherapy and psychotropic medication has increased over the past few decades, the results often fall short of expectations, 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 more than 2,000 years. Chronic sleep deprivation (CSD) induces anxiety, intestinal dysbiosis, and inflammatory responses, and reduces neurotransmitter levels, while P. cocos intervention improves anxiety-like behaviors in CSD rats, significantly increases the species richness and diversity of the gut microbiota in anxious rats, and adjusts the abundance of the dysregulated Prevotellaceae_UCG-001 to levels close to those of the normal group. Neuroinflammation is a central factor in neuropathic pain (NP). Ginger has anti-inflammatory effects and is a promising bioactive compound for NP treatment. New 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) had some effects on pain sensitivity, anxiety-like behavior, gut microbial composition and fecal metabolites in NP rats. Compared with the spinal nerve ligation (SNL) group, the SNL+GEG group and the SNL+SEG group had significantly reduced pain and anxiety-like behavior and reduced abundance of Prevotellaceae UCG-001 and Desulfovibrio.

[0003] Diabetes is a major factor endangering human health. The "Global Distribution of Diabetes" published on the website of the International Diabetes Federation (IDF) shows that there were 463 million diabetes patients worldwide in 2019, and this number is expected to reach 700 million by 2045. Type 2 diabetes (T2DM) is the most common type of diabetes, accounting for more than 90% of all diabetes diagnoses. Dyslipidemia, intestinal flora imbalance and abnormal liver lipid metabolism are the main characteristics of T2DM. Supplementation of Jerusalem artichoke inulin can improve abnormal liver lipid metabolism by changing the expression of genes involved in lipid and cholesterol production and decomposition. Microbiological analysis showed that the addition of Jerusalem artichoke inulin can enrich PrevotellaceaeUCG-001. Studies have shown that black corn is a colored corn in which anthocyanins are the most abundant bioactive compounds. 16S ribosomal RNA sequencing analysis of cecal flora showed that the intake of black corn flour increased the relative abundance of PrevotellaceaeUCG-001. Obesity is a multifactorial disease that is associated with the development of various chronic diseases. Increasing evidence shows that obesity and obesity-related metabolic diseases are largely affected by the intestinal microbiota. Fuzhuan tea is a traditional tea unique to China and has 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 richness 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. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides an intestinal microorganism that mediates the fat-protecting effect of cordycepin.

[0006] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0007] The present invention provides a role of cordycepin in the field of fat protection, and the role of cordycepin in the field of fat protection is specifically: application of cordycepin in the preparation of fat protection products in the field of treating mental diseases / sub-healthy mental states, and the therapeutic dose is 25 mg of cordycepin per kilogram of body weight.

[0008] Furthermore, the therapeutic dose is used to reduce the abundance of Prevotellaceae UCG-001 and increase the abundance of Desulfovibrio.

[0009] Furthermore, the product is a pharmaceutical composition or a food.

[0010] Furthermore, the dosage form of the pharmaceutical composition is any one of injection, tablet, capsule, suspension, ointment, microsphere, aerosol, inhalant and pill.

[0011] Furthermore, the pharmaceutical composition also includes any one or a combination of pharmaceutically acceptable excipients, carriers and matrices.

[0012] Furthermore, the food types include: health food and beverages.

[0013] Furthermore, the mental illness includes anxiety, depression, and Alzheimer's disease.

[0014] Furthermore, the sub-healthy mental state includes distraction, fatigue, anxiety, and insomnia.

[0015] Compared with the prior art, the technical solution provided by the present invention has at least the following advantages:

[0016] The present invention provides an intestinal microorganism that mediates the protective effect of cordycepin on fat. This study discusses for the first time the effects of different doses of cordycepin on anti-anxiety / depression, the changes in the richness of intestinal microorganisms, and the effects on fat weight. The present invention has been tested and simulated to prove the feasibility of the experiment. The results show that 25 mg / kg cordycepin can increase the weight reduction of white adipose tissue (sWAT) and perirenal white adipose tissue (pWAT) induced by stress, and significantly reduce the richness of Prevotellaceae UCG-001, and increase the richness of Desulfovibrio, with a trend of anti-anxiety / depression. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] One or more embodiments are exemplarily described by pictures in the corresponding drawings, and these exemplifications do not constitute limitations on the embodiments. Unless otherwise stated, the pictures in the drawings do not constitute proportional limitations.

[0018] Figure 1 This is a graph showing that cordycepin improves anxiety / depression behavior in mice according to an embodiment of the present invention;

[0019] Figure 2 is a graph of adipose tissue weights of mice in different groups in the embodiment of the present invention;

[0020] Figure 3 3 is a diagram showing the regulation of the richness of intestinal microbiota mediated by cordycepin-mediated stress damage in an embodiment of the present invention. DETAILED DESCRIPTION

[0021] The inventors found that no existing studies have linked anti-anxiety / depression to gut microbes and fat weight.

[0022] In peripheral tissues, Cpn plays an important protective and regulatory role in diabetes and adipose tissue abnormalities. Takahashi reported that Cpn exerts an anti-adipogenic effect by intervening in the activation of the C / ebpb-ppar pathway. Later studies found that high concentrations of Cpn could significantly reduce the body weight, visceral fat index, total cholesterol (TC) level, triglyceride (TG) level, low-density lipoprotein (LDL) level, and high-density lipoprotein (HDL) level of mice fed a high-fat diet; although low concentrations of Cpn could still reduce 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.

[0023] The present invention aims to explore the effects of different doses of cordycepin on anti-anxiety and depression, intestinal microorganisms and fat weight.

[0024] The present invention provides a role of cordycepin in the field of fat protection, and the role of cordycepin in the field of fat protection is specifically: application of cordycepin in the preparation of fat protection products in the field of treating mental diseases / sub-healthy mental states, and the therapeutic dose is 25 mg of cordycepin per kilogram of body weight.

[0025] Furthermore, the therapeutic dose is used to reduce the abundance of Prevotellaceae UCG-001 and increase the abundance of Desulfovibrio.

[0026] Furthermore, the product is a pharmaceutical composition or a food.

[0027] Furthermore, the dosage form of the pharmaceutical composition is any one of injection, tablet, capsule, suspension, ointment, microsphere, aerosol, inhalant and pill.

[0028] Furthermore, the pharmaceutical composition also includes any one or a combination of pharmaceutically acceptable excipients, carriers and matrices.

[0029] Furthermore, the food types include: health food and beverages.

[0030] Furthermore, the mental illness includes anxiety, depression, and Alzheimer's disease.

[0031] Furthermore, the sub-healthy mental state includes distraction, fatigue, anxiety, and insomnia.

[0032] Specifically, in the present invention, the specific testing method is:

[0033] Step 1: In the present invention, mice are first gavaged with cordycepin for 4 weeks.

[0034] Step 2: After 4 weeks, induce stress for 4 consecutive days.

[0035] Step 3. After stress, perform open field, elevated plus maze, black-white box, Y maze and tail suspension tests, collect mouse fecal samples, and weigh fat.

[0036] The following describes the 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 protection scope of the present invention is not limited to the embodiments mentioned in the text.

[0037] 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 understand the present invention more clearly and consistently, the definitions and explanations of the relevant terms are provided below.

[0038] As used herein, the terms "oral gavage" and "oral 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 an oral gavage needle.

[0039] 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 new and strange environment. The frequency and duration of certain behaviors of experimental animals in a new and strange 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, animals are afraid of new and open environments and mainly move in the peripheral areas, with less activity in the central area. However, the exploratory characteristics of 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 stimulant drugs can significantly increase autonomous activities and reduce exploratory behaviors. A certain dose of antipsychotic drugs can reduce exploratory behaviors without affecting autonomous activities.

[0040] As used herein, the term "elevated plus maze test" or "elevated plus maze" refers to the use of animals' exploratory characteristics of novel environments and their fear of high-hanging open arms to form conflicting behaviors 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 preference, 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 creates conflicting behaviors between 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.

[0041] As used herein, the term "black and white box experiment" or "light and dark box" means that in a light and dark box, mice or rats prefer to move in the dark box (black box), but the animal's exploratory habits prompt it to try to explore the light box (white box). However, the bright light stimulation of the light box inhibits the animal's exploratory activities in the light box. Anti-anxiety can remove this inhibitory effect.

[0042] As used herein, the term "Y-maze behavior experiment", "Y-maze" is generally a symmetrical closed 3-arm maze, which is shaped like a capital "Y", with an angle of 120 ° between each arm, and each arm has a movable gate to limit the activities of experimental animals. The three arms are randomly set as: novel arm, start arm and other arm. The Y-maze arm length for rats is 50 cm, the maze channel is about 10 cm wide, and the maze wall is about 13 cm high; the Y-maze arm length for mice is 28 cm, the maze channel is about 5 cm wide, and the maze wall is about 10 cm high. The entire maze is about 50 cm above the ground, and the maze is usually painted dark to prevent animals from experiencing anxiety during the experiment. The Y-maze can be used to evaluate the short-term memory of mice and rats, and also allows mice to evaluate 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 newly 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.

[0043] As used herein, the term "tail suspension test" is a classic method that can quickly evaluate the efficacy of antidepressants, stimulants, and sedatives. The principle is to use the fact that mice attempt to escape after being suspended by their tails but cannot escape, so they give up struggling and enter a unique state of depression and immobility. During the experiment, the immobility time of the animals is recorded to reflect the depressive state. Antidepressants and stimulants can significantly shorten and change their state.

[0044] The present invention will be further described below through specific examples.

[0045] The experimental mice were subjected to a 4-day repeated stress protocol, which included 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:

[0046] Control + water: no stress + pure water gavage;

[0047] Stress + water: stress + pure water gavage;

[0048] Stress+12.5Cpn: stress+12.5mg / kg cordycepin gavage;

[0049] Stress+25Cpn: stress+25mg / kg cordycepin gavage;

[0050] No stress + 25Cpn: No stress + 25 mg / kg cordycepin administered orally.

[0051] After 4 weeks of oral administration, open field test (OFT), elevated plus maze (EPM), black-and-white box (LDB), Y-maze test (Y-maze) and tail suspension test (TST) were performed. The feces of mice were collected for 16S ribosomal RNA sequencing, and the subcutaneous white adipose tissue (sWAT), perirenal white adipose tissue (pWAT), epididymal white adipose tissue (eWAT) and brown adipose tissue (BAT) of mice were weighed.

[0052] Figure 1 This is a graph showing that cordycepin improves anxiety / depression behavior in mice according to an embodiment of the present invention. Figure 1In the figure, (A) is the movement trajectory, intermediate residence time, total moving distance, intermediate distance / total distance, and intermediate entry times of mice in OFT; (B) is the movement trajectory, open arm residence time, total static time, open arm entry times, and open arm movement distance of mice in EPM test; (C) is the latency time, white box residence time, and white box / black box crossing times of LDB test; (D) is the number of spontaneous alternations and total arm entry times of Y-maze test; (E) is the static time of TST. P values ​​were analyzed by unpaired t-test or one-way ANOVA, and the rest of the groups were analyzed by 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: stress + pure water gavage: stress + 12.5Cpn: stress + 12.5mg / kg cordycepin gavage: stress + 25Cpn: stress + 25mg / kg cordycepin gavage: no stress + 25Cpn: no stress + 25mg / kg cordycepin gavage. OFT: open field test; EPM: elevated plus maze; LDB: black and white box; Y-maze: y maze; TST: tail suspension test.

[0053] Figure 2 It is a graph of the adipose tissue weight of mice in different groups in the embodiment of the present invention. #p<0.05, **p<0.01, ##p<0.01, determined by unpaired t-test or one-way ANOVA Tukey multiple comparison test. The bar graph represents the mean ± SEM value. N=5 per group. Control + water group: no stress + pure water gavage; stress + water: stress + pure water gavage: stress + 12.5Cpn: stress + 12.5mg / kg cordycepin gavage: stress + 25Cpn: stress + 25mg / kg cordycepin gavage: no stress + 25Cpn: no stress + 25mg / kg cordycepin gavage. pWAT: perirenal white adipose tissue, eWAT: epididymal white adipose tissue, sWAT: subcutaneous white adipose tissue, BAT: brown adipose tissue.

[0054] Figure 3 is a diagram showing the regulation of the richness of intestinal microbiota mediated by cordycepin-mediated stress damage in an embodiment of the present invention. Figure 3(A) is the relative abundance of Prevotellaceae_UCG-001, (B) is NK4A214_group, and (C) is Desulfovibrio. #p<0.05, *P<0.05, **P<0.01, determined by unpaired t-test or one-way ANOVA Tukey multiple comparison test. Bar graphs represent mean ± SEM values. N=4-5 per group. Control + water group: no stress + pure water gavage; stress + water: stress + pure water gavage: stress + 12.5Cpn: stress + 12.5mg / kg cordycepin gavage: stress + 25Cpn: stress + 25mg / kg cordycepin gavage: no stress + 25Cpn: no stress + 25mg / kg cordycepin gavage.

[0055] In summary, although there was no significant difference between 25mg / kg cordycepin and the stress group, it had an anti-anxiety / depression trend. 25mg / kg cordycepin can significantly increase the weight reduction of white adipose tissue (sWAT) and perirenal white adipose tissue (pWAT) caused by stress, but has no effect on the weight of epididymal white adipose tissue (eWAT) and brown adipose tissue (BAT).

[0056] The present invention has been tested and simulated to prove the feasibility of the experiment. The results show that 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, and increase the abundance of Desulfovibrio, with an anti-anxiety / depression trend.

[0057] Those skilled in the art can understand that the above-mentioned embodiments are specific examples for implementing the present application, and in practical applications, various changes can be made to them in form and detail without departing from the spirit and scope of the present application. Any person skilled in the art can make their own changes and modifications without departing from the spirit and scope of the present application, so the scope of protection of the present application shall be based on 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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