Application of ginsenoside CK in treatment of insomnia

Abstract

The invention discloses application of ginsenoside CK or salt thereof in preparation of drugs, food and / or health care products for preventing and / or treating insomnia. Ginsenoside CK shows the most significant anti-inflammatory and insomnia-improving effects in vitro and in vivo.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to application of ginsenoside CK in treating insomnia. Background Art

[0002] Insomnia is a common sleep disorder, which means that despite having a good sleeping environment and sufficient sleep opportunities, individuals are still dissatisfied with the quality or duration of sleep, and their daily life and social functions are affected. Clinically, insomnia can be divided into short-term insomnia (lasting no more than three months) and chronic insomnia (lasting more than three months). Its main manifestations include sleep maintenance disorders such as difficulty falling asleep, easy awakening, and early awakening, and are often accompanied by daytime fatigue, sleepiness, emotional tension, anxiety, irritability and other symptoms. Epidemiological surveys show that the prevalence of insomnia in adults is relatively high, especially women, the elderly and people facing socioeconomic pressures are more susceptible. Insomnia tends to be chronic and has a high persistence rate.

[0003] At present, the commonly used chemical drugs for the clinical treatment of insomnia include melatonin and its receptor agonists, benzodiazepines, Benzodiazepines and non-benzodiazepines There are many types of insomnia, such as sedatives, barbiturates, and certain antidepressants, antipsychotics, and antihistamines with sedative effects. Although these drugs have certain efficacy in the treatment of short-term insomnia, they are prone to safety risks during long-term use, such as drug tolerance, dependence, and unstable efficacy, which limits their application in the management of chronic insomnia. Therefore, finding safer and more effective treatment strategies has become a research focus. Among them, traditional Chinese medicine, due to its overall regulatory effect and low side effects, has shown unique advantages in the field of insomnia treatment and broad prospects for new drug research and development.

[0004] As a common medicine in traditional Chinese medicine, ginseng Chinese medicinal materials occupy an important position in the prescriptions for the treatment of insomnia. Classic prescriptions such as Guipi Decoction and Tianwang Buxin Dan are often used to treat insomnia caused by deficiency of both heart and spleen, insufficient qi and blood, etc. They may have certain biological activities in the nervous system, such as regulating neurotransmitter balance, anti-inflammatory, antioxidant and neuroprotective effects. However, further research and verification are needed on their specific anti-neuroinflammatory mechanism and their role in pathological conditions related to insomnia.

[0005] The prior art does not disclose the use of ginsenoside CK as claimed in the present invention for treating insomnia. Summary of the invention

[0006] Based on this, the present invention provides the use of ginsenoside CK or its salt in the preparation of medicines, foods and / or health products for preventing and / or treating insomnia.

[0007] According to another aspect of the present invention, there is provided a use of a composition comprising ginsenoside CK or a salt thereof in the preparation of medicines, foods and / or health products for preventing and / or treating insomnia.

[0008] Furthermore, the ginsenoside CK prevents and / or treats insomnia in one or more of the following ways: inhibiting the secretion of inflammatory factors in cells in vitro, improving abnormal behavior, improving sleep quality, alleviating anxious behavior, and inhibiting the secretion of inflammatory factors in organisms.

[0009] Furthermore, the cell is a BV-2 cell.

[0010] Furthermore, the cellular inflammatory factors include one or more of the following: NO, IL-6 and TNF-α.

[0011] Furthermore, the behavioral abnormality includes an increase in the amount of wakefulness activity and / or an increase in wakefulness time.

[0012] Further, the improvement of sleep quality includes shortening sleep latency and / or prolonging sleep time.

[0013] Furthermore, the biological inflammatory factors include one or more of the following: IL-6, TNF-α and IL-1β.

[0014] Furthermore, the composition further comprises one or more pharmaceutically acceptable excipients.

[0015] Furthermore, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, glidant, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0016] Furthermore, the dosage form of the composition is powder, tablet, pill, capsule, film, lozenge, granule, injection or oral solution.

[0017] Furthermore, the composition further comprises one or more other drugs and / or extracts for treating insomnia.

[0018] Further, the other drugs are selected from one or more of the following: benzodiazepines Receptor agonists, antidepressants, and melatonin receptor agonists.

[0019] Furthermore, the benzodiazepine Benzodiazepines Drugs and / or non-benzodiazepines Class of drugs.

[0020] Furthermore, the benzodiazepine The drugs in this category are estazolam, triazolam, and / or diazepam.

[0021] Furthermore, the non-benzodiazepine The drugs in this class are zolpidem, zopiclone, and / or eszopiclone.

[0022] Furthermore, the antidepressant drugs include noradrenergic and specific 5-HTergic antidepressants and / or 5-HT receptor blockers and reuptake inhibitors.

[0023] Furthermore, the noradrenergic and specific 5-HTergic antidepressant drug is mirtazapine.

[0024] Furthermore, the 5-HT receptor blocker and reuptake inhibitor is trazodone.

[0025] Furthermore, the melatonin receptor agonist is agomelatine and / or ramucirumab.

[0026] Beneficial effects of the present invention:

[0027] The present invention first compares the inhibitory effects of ginsenoside CK, Rg1, Rb1, RC, notoginsenoside R1, notoginsenoside K and notoginseng on LPS-induced inflammatory response in BV2 cells at different concentrations (0.2 μM, 1 μM, 5 μM, 25 μM), and screens out components with significant anti-inflammatory effects. Subsequently, multiple dose experiments were carried out in zebrafish and mouse insomnia models to evaluate their in vivo effectiveness. The results showed that ginsenoside CK exhibited the most significant anti-inflammatory and insomnia-improving effects both in vitro and in vivo. DETAILED DESCRIPTION

[0028] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0029] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those of ordinary skill in the field of the invention or the field in which the terms are used. Although any methods, conditions, substances or materials similar to or equivalent to those disclosed herein may be used in the practice of the present invention, preferred methods, conditions, substances or materials are described herein.

[0030] The present invention is intended to encompass all alternatives, variations and equivalents that may be included in the present invention as defined by the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein that can be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[0031] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0032] In the present invention, the term "comprising" is synonymous with "including". As used herein, the terms "comprising", "including", "having", "containing" or any other variation thereof are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus comprising the listed elements is not necessarily limited to only those elements, but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

[0033] As described in the background technology section, the prior art does not disclose the use of ginsenoside CK as claimed in the present invention for treating insomnia. In order to solve the above problems, the present invention provides the use of ginsenoside CK or its salt in the preparation of medicines, foods and / or health products for preventing and / or treating insomnia.

[0034] According to another aspect of the present invention, there is provided a use of a composition comprising ginsenoside CK or a salt thereof in the preparation of medicines, foods and / or health products for preventing and / or treating insomnia.

[0035] In a preferred embodiment, the ginsenoside CK prevents and / or treats insomnia in one or more of the following ways: inhibiting the secretion of inflammatory factors in cells in vitro, improving abnormal behavior, improving sleep quality, alleviating anxiety behavior, and inhibiting the secretion of inflammatory factors in organisms.

[0036] In a preferred embodiment, the cell is a BV-2 cell.

[0037] In a preferred embodiment, the cellular inflammatory factors include one or more of the following: NO, IL-6 and TNF-α.

[0038] In a preferred embodiment, the behavioral abnormality includes increased wakefulness activity and / or increased wakefulness time.

[0039] In a preferred embodiment, the improvement of sleep quality includes shortening sleep latency and / or prolonging sleep time.

[0040] In a preferred embodiment, the biological inflammatory factors include one or more of the following: IL-6, TNF-α and IL-1β.

[0041] In a preferred embodiment, the composition further comprises one or more pharmaceutically acceptable excipients.

[0042] In a preferred embodiment, the auxiliary material is selected from one or more of the following: diluents, wetting agents, binders, disintegrants, inclusion agents, flavoring agents, sustained-release agents, glidants, lubricants, dispersants, plasticizers, opacifiers and antioxidants.

[0043] In a preferred embodiment, the composition is in the form of powder, tablet, pill, capsule, film, lozenge, granule, injection or oral solution.

[0044] In a preferred embodiment, the composition further comprises one or more other drugs and / or extracts for treating insomnia.

[0045] In a preferred embodiment, the other drug is selected from one or more of the following: benzodiazepines Receptor agonists, antidepressants, and melatonin receptor agonists.

[0046] In a preferred embodiment, the benzodiazepine Benzodiazepines Drugs and / or non-benzodiazepines Class of drugs.

[0047] In a preferred embodiment, the benzodiazepine The drugs in this category are estazolam, triazolam, and / or diazepam.

[0048] In a preferred embodiment, the non-benzodiazepine The drugs in this class are zolpidem, zopiclone, and / or eszopiclone.

[0049] In a preferred embodiment, the antidepressant drugs include noradrenergic and specific 5-HTergic antidepressants and / or 5-HT receptor blockers and reuptake inhibitors.

[0050] In a preferred embodiment, the noradrenergic and specific 5-HTergic antidepressant is mirtazapine.

[0051] In a preferred embodiment, the 5-HT receptor blocker and reuptake inhibitor is trazodone.

[0052] In a preferred embodiment, the melatonin receptor agonist is agomelatine and / or ramucirumab.

[0053] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually carried out according to conventional conditions or conditions recommended by the manufacturer.

[0054] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0055] The above features mentioned in the present invention or the features mentioned in the embodiments can be combined in any way. All the features disclosed in this patent specification can be used in combination with any combination form, and each feature disclosed in the specification can be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features.

[0056] Example

[0057] 1. Experimental instruments, materials and methods

[0058] 1.1 Experimental instruments, reagents and materials

[0059] Ginsenoside CK, Rg1, Rb1, RC, notoginsenoside R1, K, notoginsenoside (purity ≥ 98%, purchased from Chengdu Aifa Biotechnology Co., Ltd.); lipopolysaccharide (LPS, Escherichiacoli055:B5) (Sigma-Aldrich, USA); CCK-8 cell proliferation and toxicity detection kit (Sevier Biotechnology Co., Ltd.); ELISA kit (detection of IL-6, TNF-α, IL-1β) (Sevier Biotechnology Co., Ltd.); DMEM high glucose medium, fetal bovine serum (FBS), trypsin (Gibco, USA); DMSO, PBS, phenol red (Sigma- Aldrich, USA); main instruments and equipment cell culture incubator (ThermoFisher, 37℃, 5% CO2); inverted microscope (Nikon TS100); microplate reader (BioTek Synergy H1, USA); PCR instrument (ThermoFisher Quant Studio 3); fluorescence microscope (Leica DMi 8); high performance liquid chromatograph (ThermoFisher Scientific); CO2 incubator (ThermoForma 3111); ultra-low temperature refrigerator (-80℃) (Haier DW-86L486).

[0060] 1.2 Cell culture

[0061] Mouse microglial cells BV2 (provided by Wuhan Punosai); culture medium: DMEM (containing 10% FBS, 1% penicillin-streptomycin); culture conditions: 37°C, 5% CO2.

[0062] 1.3 Establishment of Cellular Inflammation Model

[0063] 1×10 4 BV-2 cells were inoculated at a density of 100 μg / mL in a 96-well plate, cultured for a period of time and observed under a microscope. After the cells were completely attached to the wall, the original culture medium was removed and replaced with LPS solutions of different concentrations (0.1 μg / mL, 1 μg / mL, 10 μg / mL, 100 μg / mL LPS) and incubated for 24 hours for CCK8 and NO determination. NO determination: After LPS modeling, the cell supernatant was collected and centrifuged for 5 minutes, 50 μL of the supernatant from each group was drawn and placed in a 96-well plate, 50 μL of GriessI and 50 μL of GriessII solution were added to each well, and the absorbance was measured at 540 nm after 5 minutes of reaction. The data were imported into Excel, and the NO concentration was calculated according to the standard curve.

[0064] 1.4. Zebrafish Insomnia Model Construction and Drug Administration

[0065] To further verify the anti-insomnia and anti-inflammatory effects of the monomer that significantly improved inflammation in in vitro experiments in the in vivo model, a zebrafish insomnia model was induced by immersion administration of 5mM pentylenetetrazol (PTZ), and the intervention doses for zebrafish (0.125, 0.623, and 3.114 μg / mL) were converted according to the cell experimental doses.

[0066] 1.5. PCPA insomnia model mouse model establishment, drug administration, and sample preparation

[0067] C57BL / 6 mice (male, 18–22 g) were selected and adaptively fed for 7 days and randomly divided into a normal control group (Normal), a model group (PCPA), and a drug intervention group (CK, Rg1, Rb1), with n = 9 in each group. PCPA was dissolved in 0.9% NaCl and injected intraperitoneally at 300 mg / kg body weight for 3 consecutive days to induce a mouse insomnia model. After the modeling was completed, the mice were gavaged daily (CK, Rg1, Rb1) for 11 consecutive days. The dosage was administered with reference to in vitro experiments and equivalent conversion. The normal control group and the model group were gavaged with an equal volume of saline. Behavioral tests were performed 1 hour after the last administration. The enzyme-linked immunosorbent assay (ELISA) method was used to detect the IL-6 level in the mouse brain tissue to evaluate the regulatory effect of ginsenoside CK on neuroinflammation in PCPA-induced insomnia model mice. Preparation of brain tissue homogenate: After the mice were killed, the hippocampus and prefrontal cortex tissues were quickly removed and washed with pre-cooled PBS (pH 7.4) saline to remove blood. Take 50 mg of tissue, add 500 μL of RIPA lysis buffer, and homogenize with a tissue homogenizer in an ice bath for 30 seconds, with an interval of 10 seconds, for a total of 3 times. Centrifuge the homogenate at 4°C, 12,000 rpm for 10 minutes, and take the supernatant for later use.

[0068] 1.6. NO detection

[0069] In a 96-well plate, 2 × 10 5 BV-2 cells were inoculated at a density of 1 μg / mL, cultured for a period of time and observed under a microscope, the original culture medium was removed, and replaced with serum-free culture medium containing 0.2, 1, 5, and 25 μM of each monomer component, pre-protected for 2 hours, and then LPS (1 μg / mL) was added and cultured for 24 hours. According to the NO detection method in 1.2, 50 μL of the supernatant was aspirated into a 96-well plate, the working solution was added and shaken to mix, and the absorbance was measured at 540 nm using an ELISA reader.

[0070] 1.7. Detection of inflammatory factors

[0071] The supernatant of each group was collected and centrifuged, and the ELISA kit was used for determination. The absorbance value was measured by a microplate reader, and the standard curve was made by importing Excel according to the instruction manual to calculate the content of IL-6, TNF-α, and IL-1β in the supernatant.

[0072] 2. Experimental results

[0073] 2.1. Establishment of LPS-induced BV2 cell inflammation model

[0074] The damage concentration of LPS-induced microglial inflammation model was explored by measuring NO levels. The results showed that LPS had no effect on cell viability after 24h treatment with 0.01μg / mL, 1μg / mL, 10μg / mL and 100μg / mL, while 1μg / mL, 10μg / mL and 100μg / mL LPS significantly increased the release of NO in a concentration-dependent manner, indicating that 1μg / ml LPS can induce BV-2 cell inflammation. Therefore, LPS with a concentration of 1μg / mL was selected for subsequent experiments. The experimental results are shown in Tables 1 and 2.

[0075] Table 1 Effect of LPS on BV-2 cell viability (n=6, )

[0076]

[0077]

[0078] Table 2 Effects of LPS on NO levels in BV-2 cells (n=6, )

[0079] LPS (μg / mL) NO production (μM) 0 0.242±0.161 0.01 0.361±1.667 0.1 0.686±0.098 1 1.985±0.269** 10 2.496±0.569** 100 2.975±0.598**

[0080] Significance: Compared with the control group, * P<0.05, ** P<0.01.

[0081] 2.2. CCK8 cell viability assay

[0082] The results of CCK8 showed that ginsenoside CK, ginsenoside Rg1, ginsenoside Rb1, ginsenoside RC, notoginsenoside K, notoginsenoside R1, and notoginseng were not toxic to BV-2 cells at 0.2 μM, 1 μM, 5 μM, and 25 μM. The experimental results are shown in Table 3.

[0083] Table 3 Effects of various monomer components on the survival rate of BV2 cells (n=6, )

[0084] Group 0μM 0.2μM 1μM 5μM 25μM LPS 98.85±1.33 97.88±1.92 97.45±2.11 97.32±1.84 96.21±2.34 LPS+CK 101.16±1.74 99.62±1.73 98.85±1.91 98.21±1.89 94.15±3.02 LPS+Rg1 101.23±2.68 99.45±1.80 98.92±1.85 96.89±2.07 92.21±3.56 LPS+Rb1 99.22±1.97 99.72±1.66 99.01±1.74 97.62±1.95 94.93±3.27 LPS+K 100.01±1.20 99.81±1.69 99.12±1.78 98.03±2.13 91.21±2.89 LPS+Notoginseng 99.03±1.49 99.58±1.89 99.18±1.43 96.98±1.48 93.83±2.37 LPS+R1 97.59±0.91 99.41±1.76 98.25±1.56 94.04±2.65 92.91±2.71 LPS+RC 100.50±2.10 98.88±1.45 98.06±1.64 97.03±2.84 93.31±3.76

[0085] 2.3 Effects of each monomer component on the levels of NO, IL-6, and TNF-α in BV-2 cells induced by LPS The experiment evaluated the effects of ginsenoside CK (CK), ginsenoside Rg1 (Rg1), ginsenoside Rb1 (Rb1), ginsenoside RC (RC), notoginseng saponin K (K), notoginseng saponin R1 (R1), and dencichine on the release of NO and the levels of IL-6 and TNF-α in BV-2 cells induced by LPS. The results showed that the LPS group (1 μg / mL) significantly increased the secretion levels of NO, IL-6, and TNF-α (P<0.01), indicating that the inflammatory model was successfully established. Ginsenoside CK (CK) can significantly inhibit the secretion of TNF-α, IL-6, and NO at concentrations of 0.2-25 μM, among which the 5 μM and 25 μM groups had the strongest effects (P<0.01). Ginsenoside Rg1 (Rg1) and ginsenoside Rb1 (Rb1) can significantly reduce TNF-α and NO at higher concentrations (≥5μM) (P<0.05), and have a weaker effect on IL-6. Ginsenoside RC (RC), notoginsenoside K (K), notoginsenoside R1 (R1), and dencichine have no significant inhibitory effect on the secretion of NO, IL-6, and TNF-α in the concentration range of 0.2-25μM (P>0.05). The experimental results are shown in Table 4.

[0086] Table 4 Effects of various monomer components on LPS-induced NO, IL-6, and TNF-α levels in BV-2 cells (n=6, )

[0087]

[0088]

[0089] Significance: Compared with the normal group, # P<0.05, ## P<0.01, comparison between the test group and the model group, * P<0.05, **P<0.01.

[0090] 2.4 Behavioral verification of the zebrafish insomnia model

[0091] The results showed that the PTZ group (5mM) significantly increased the amount of awakening activity (mm) of zebrafish (P<0.01), indicating that the insomnia model was successfully established. Ginsenoside CK (CK) significantly reduced the amount of awakening activity of zebrafish at all concentrations (0.125-3.114μg / mL) (P<0.05 or P<0.01), and showed a dose-dependent trend to improve insomnia. Ginsenoside Rg1 (Rg1) and ginsenoside Rb1 (Rb1) had no significant effect at the above concentrations. The experimental results are shown in Table 5.

[0092] Table 5 Effects of ginsenoside CK, ginsenoside Rg1, and ginsenoside Rb1 on the behavior of zebrafish insomnia model (n=10, )

[0093]

[0094]

[0095] Significance: Compared with the normal group, ## P<0.01, comparison between the test group and the model group, * P<0.05, ** P<0.01.

[0096] 2.5 In vivo experimental verification of mouse insomnia model

[0097] In order to further verify the monomers with significant effects on improving insomnia in zebrafish experiments, this study used PCPA-induced 5-HT-deficient insomnia mouse model for experiments. According to the conversion formula from cell experiments to in vivo experiments, the mouse dosage (0.01 mg / kg, 0.05 mg / kg, 0.25 mg / kg) was determined, and intervention experiments were carried out. The results showed that ginsenoside CK could significantly shorten the sleep latency in each dose group (P<0.01) and significantly reduce the level of IL-6, indicating that it can effectively improve the symptoms of PCPA-induced insomnia and show a certain dose dependence. In addition, ginsenoside Rg1 showed a certain effect of shortening the sleep latency in the high concentration group (0.25 mg / kg) (P<0.05), while ginsenoside Rb1 showed no significant changes; ginsenoside Rb1 showed a certain effect of reducing the level of IL-6 in the high concentration group (0.25 mg / kg) (P<0.05), while ginsenoside Rg1 showed no significant changes. The experimental results are shown in Tables 6 and 7.

[0098] Table 6 Effects of ginsenoside CK, ginsenoside Rg1, and ginsenoside Rb1 on the behavior of mice (n=9, )

[0099]

[0100]

[0101] Significance: Compared with the normal group, ## P<0.01, comparison between the test group and the model group, *P<0.05, **P<0.01.

[0102] Table 7 Effects of ginsenoside CK, ginsenoside Rg1, and ginsenoside Rb1 on IL-6 levels in mice (n=9, )

[0103] Group IL-6 (pg / mL) Normal 14.872±2.315 PCPA (300 mg / kg) <![CDATA[47.315±3.928 ## <!-- 9 -->]]> PCPA+CK (0.01 mg / kg) <![CDATA[38.412±3.215 * ]]> PCPA+CK (0.05 mg / kg) <![CDATA[36.287±2.978 * ]]> PCPA+CK (0.25 mg / kg) <![CDATA[25.914±2.462 ** ]]> PCPA+Rg1(0.01mg / kg) 44.873±3.728 PCPA+Rg1(0.05mg / kg) 42.315±3.591 PCPA+Rg1(0.25mg / kg) 40.214±3.102 PCPA+Rb1(0.01mg / kg) 45.210±3.862 PCPA+Rb1(0.05mg / kg) 43.762±3.648 PCPA+Rb1(0.25mg / kg) <![CDATA[37.824±3.218 * ]]>

[0104] Significance: Compared with the normal group, ## P<0.01, comparison between the test group and the model group, * P<0.05, ** P<0.01.

[0105] The embodiments of the present invention are described in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only used to help understand the method of the present invention and its core idea. At the same time, changes or deformations made by those skilled in the art based on the ideas of the present invention, the specific implementation methods and application scope of the present invention, all belong to the scope of protection of the present invention. In summary, the content of this specification should not be understood as limiting the present invention.

Claims

1. Use of ginsenoside CK or its salt in the preparation of medicines, foods and / or health products for preventing and / or treating insomnia.

2. Use of a composition comprising ginsenoside CK or a salt thereof in the preparation of medicines, foods and / or health products for preventing and / or treating insomnia.

3. The use according to claim 2 or 3, characterized in that: The ginsenoside CK prevents and / or treats insomnia in one or more of the following ways: inhibiting the secretion of in vitro cellular inflammatory factors, improving abnormal behavior, improving sleep quality, alleviating anxiety behavior, and inhibiting the secretion of inflammatory factors in the organism.

4. The use according to claim 3, characterized in that The cells are BV-2 cells; Preferably, the cellular inflammatory factors include one or more of the following: NO, IL-6 and TNF-α; Preferably, the behavioral abnormality includes increased amount of wakefulness activity and / or increased wakefulness time; Preferably, the improvement of sleep quality includes shortening sleep latency and / or prolonging sleep time; Preferably, the biological inflammatory factors include one or more of the following: IL-6, TNF-α and IL-1β.

5. The use according to claim 2, characterized in that: The composition further comprises one or more pharmaceutically acceptable excipients.

6. The use according to claim 5, characterized in that The auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, glidant, lubricant, dispersant, plasticizer, opacifier and antioxidant.

7. The use according to claim 2, characterized in that: The dosage form of the composition is powder, tablet, pill, capsule, film, lozenge, granule, injection or oral solution.

8. The use according to claim 2, characterized in that: The composition further comprises one or more other drugs and / or extracts for treating insomnia.

9. The use according to claim 8, characterized in that The other drugs are selected from one or more of the following: benzodiazepines Receptor agonists, antidepressants, and melatonin receptor agonists.

10. The use according to claim 9, characterized in that The benzodiazepine Receptor agonists including benzodiazepines Drugs and / or non-benzodiazepines Class drugs; Preferably, the benzodiazepine The drugs in this category are estazolam, triazolam, and / or diazepam; Preferably, the non-benzodiazepine The drugs in this category are zolpidem, zopiclone, and / or eszopiclone; More preferably, the antidepressant drugs include noradrenergic and specific 5-HTergic antidepressants and / or 5-HT receptor blockers and reuptake inhibitors; Also preferably, the noradrenergic and specific 5-HT antidepressant is mirtazapine; Also preferably, the 5-HT receptor blocker and reuptake inhibitor is trazodone; Also preferably, the melatonin receptor agonist is agomelatine and / or ramucirumab.

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