Composition with sleep improving effect and application thereof

Abstract

The invention belongs to the technical field of pharmaceutical compositions, and particularly relates to a composition with a sleep improving effect and application thereof. The composition comprises gamma-aminobutyric acid and a poria cocos extract, can also comprise a rhizoma polygonati extract and phosphatidylserine, and can achieve the effect of improving sleep within a specific ratio range. The composition has both functionality and nutrition, and shows a wide application prospect in the aspect of improving the sleep function.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical compositions, and particularly relates to a composition capable of improving sleep and application thereof. Background Art

[0002] Sleep is an essential natural physiological process that promotes the removal of neurotoxic waste and maintains synapses related to memory and learning. Therefore, sleep is essential for brain function, especially cognitive function. Insomnia is associated with various body dysfunctions, including endocrine, metabolic, immune, cardiovascular, cortical and nervous system disorders. Short-term insomnia can affect memory and attention, while long-term insomnia can lead to neurological dysfunction, exacerbation of existing diseases, and even death in severe cases. Insomnia has become a common concern, affecting people's physical and mental health.

[0003] Traditional hypnotic drugs increase the total sleep time at night and reduce sleep latency, but there are risks such as easy overdose, increased tolerance and addiction, and inducing daytime sedation and withdrawal reactions. As a public health problem, insomnia has not yet been fully and effectively treated. Gamma-aminobutyric acid (GABA) is an important inhibitory neurotransmitter that is mainly found in the central nervous system. Its main function is to regulate the excitability of neurons and help balance brain activity. In recent years, studies have found that GABA can promote relaxation and calmness by binding to its receptors, thereby helping to reduce anxiety and stress, and is also closely related to the improvement of sleep quality.

[0004] Poria extract contains a variety of active ingredients, such as polysaccharides, triterpenoids, etc. These ingredients work together to help improve sleep quality. In clinical applications, many patients reported that their sleep time was extended after taking Poria extract, and they felt energetic and happy when they woke up.

[0005] Although GABA shows some potential in improving sleep, there are some limitations to its functional effects when used alone. On the one hand, due to the existence of the human blood-brain barrier, after oral administration, the amount of GABA that can effectively pass through the blood-brain barrier and enter the brain to play a role is relatively limited, making it difficult to achieve the desired ideal state in improving sleep. On the other hand, when GABA is used alone, its duration of action is short, and it is difficult to achieve a lasting good sleep state. Therefore, it is necessary to introduce other substances to work together with GABA to improve sleep, which is necessary for the development of products with better sleep improvement effects and better nutritional functions. Summary of the invention

[0006] In view of the shortcomings of the prior art, the present invention aims to provide a composition that helps improve sleep and its application, which can achieve a better effect of improving sleep by rationally compounding GABA with other functional components. The composition takes into account both functionality and nutrition, and shows broad application prospects in improving sleep function.

[0007] To achieve the above objectives, on the one hand, the present invention provides a composition that helps to improve sleep, the composition comprising γ-aminobutyric acid and Poria cocos extract, the mass ratio of the γ-aminobutyric acid to the Poria cocos extract being 55-85:15-45, for example 85:15, 70:30, 55:45, 66:45 or 77:23.

[0008] Furthermore, the mass ratio of the γ-aminobutyric acid to the Poria cocos extract is 11:9.

[0009] In a second aspect, the present invention provides a composition that helps to improve sleep, the composition comprising γ-aminobutyric acid and a Poria cocos extract, the composition also comprising a Polygonatum sibiricum extract and phosphatidylserine, the weight ratio of the γ-aminobutyric acid, Poria cocos extract, Polygonatum sibiricum extract and phosphatidylserine being 55-85:15-45:30-40:2-15, for example, 85:15:40:5, 70:30:40:5, 55:45:40:5, 66:45:40:5, 77:23:40:5, 55:45:30:15, 55:45:43:2.

[0010] Furthermore, the weight ratio of the γ-aminobutyric acid, the Poria cocos extract, the Polygonatum sibiricum extract and the phosphatidylserine is 11:9:8:1.

[0011] In a third aspect, the present invention provides a product, comprising the composition described in the first aspect or the second aspect.

[0012] Furthermore, the product is a health food or a medicine, and the product also includes excipients acceptable to health food or medicine.

[0013] In a fourth aspect, the present invention provides a use of the composition described in the first aspect in preparing a product having a sleep-improving effect, wherein the product is a health food or a medicine.

[0014] Furthermore, the application is an application in preparing a product capable of prolonging sleep time. Furthermore, the application is one or more of the following: (1) Application in the preparation of products capable of improving the decreased GABA secretion level caused by insomnia; (2) Application in the preparation of products capable of improving the decrease in 5-HT levels caused by insomnia; (3) Application in the preparation of products capable of improving the increase of ACTH content caused by insomnia; (4) Application in the preparation of products capable of improving the increase in CORT content caused by insomnia; (5) Application in the preparation of products capable of improving oxidative damage caused by insomnia; (6) Use in the preparation of products capable of improving increased inflammation levels caused by insomnia; (7) Use in the preparation of products capable of improving neuronal cell damage caused by insomnia.

[0015] Compared with the prior art, the present invention has the following advantages: (1) The present invention proves that the combined intervention of γ-aminobutyric acid and Poria cocos extract can help improve sleep, and the two components have a synergistic effect; (2) The present invention proves that the combined intervention of γ-aminobutyric acid, Poria cocos extract, Polygonatum sibiricum extract and phosphatidylserine can help improve sleep, and the four components have a synergistic effect.

[0016] (3) The composition of the present invention can improve sleep, prolong the sleep time of insomnia patients, and has the functions of improving the decrease in GABA secretion level caused by insomnia, improving the decrease in 5-HT level caused by insomnia, improving the increase in ACTH content caused by insomnia, improving the increase in CORT content caused by insomnia, improving the oxidative damage caused by insomnia, improving the increase in inflammation level caused by insomnia, and improving the damage to neuronal cells caused by insomnia. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 Shown are the results of the effect of the composition on the sleep latency of mice induced by sodium pentobarbital.

[0018] Figure 2 Shown are the results of the effects of the composition on the sleep duration of mice induced by sodium pentobarbital.

[0019] Figure 3 The figure shows the effect of the composition on the GABA content and 5-HT content in the serum and brain of mice, wherein A is the serum GABA content; B is the brain GABA content; C is the serum 5-HT content; and D is the brain 5-HT content.

[0020] Figure 4 The figure shows the effect of the composition on serum ACTH content and CORT content in mice, wherein A is serum ACTH content; B is serum CORT content.

[0021] Figure 5The figure shows the effect of the composition on the levels of serum SOD, CAT and GSH in mice, wherein A is the serum SOD content; B is the serum CAT content; and C is the serum GSH content.

[0022] Figure 6 The figure shows the effect of the composition on the levels of TNF-α, IL-1β and IL-6 in serum of mice, wherein A is the level of serum TNF-α; B is the level of serum IL-1β; and C is the level of serum IL-6.

[0023] Figure 7 Shown is the effect of the composition on the histopathological examination results of the mouse hippocampus.

[0024] Figure 8 Shown is the effect of the composition on the density of c-Fos-positive cells in the mouse hippocampus. DETAILED DESCRIPTION

[0025] The following is a further description of the concept of the present invention and the technical effects produced in conjunction with specific embodiments, so as to fully understand the purpose, features and effects of the present invention. The methods are conventional methods unless otherwise specified. The materials can be obtained from public commercial channels unless otherwise specified. The illustrative embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations of the present invention. It should be noted that the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

[0026] Example 1 Screening of composition raw materials GABA and 5-HT are important neurotransmitters in the central nervous system and play a key role in regulating the sleep-wake cycle and improving sleep quality. The present invention uses the expression levels of GABA mRNA and 5-HT mRNA to characterize the potential activity of each substance in improving sleep. The specific steps are as follows: Each test substance was dissolved in DMEM medium (containing 10% fetal bovine serum and 1% penicillin-streptomycin) and filtered through a 0.22 µm sterile syringe filter to prepare a solution with a concentration of 250-2000 µg / mL. SH-SY5Y cells were cultured at 5×10 4 The cells were seeded at a density of 1.54 × 10 / mL in a 6-well plate and incubated at 37°C with 5% CO 2 After overnight culture under the same conditions and observation of cell adhesion, the original culture medium was discarded in each intervention group, and 3 mL of the test substances with different components were used to intervene in SH-SY5Y cells, while the blank control group continued to be cultured with the original culture medium. 37 ℃, 5% CO 2After 48 h of culture under the same conditions, the medium in the 6-well plate was discarded, 1 mL of Trizol was added to each well, and the solution in each well was transferred to a 1.5 mL Eppendorf tube. 200 μL of chloroform was added, and the solution was shaken until it was uniformly milky white. Centrifuged at 4 °C, 12000 rpm for 15 min, and the upper aqueous phase was transferred to another clean centrifuge tube. An equal volume of isopropanol was added and mixed by inversion. After standing at room temperature for 30 min, centrifuged at 4 °C, 12000 rpm for 15 min. The supernatant was discarded, and 1 mL of -20 °C pre-cooled 75% ethanol (prepared with DEPC water) was added to wash the precipitate (the precipitate was white); centrifuged at 4 °C, 5000 rpm for 5 min, the supernatant was carefully discarded, the precipitate was dried, dissolved in 30 μL of DEPC water, and frozen at -80 °C.

[0027] Take 10 μL RNA in a 200 μL enzyme-free EP tube, add 4 μL 5×All-In-One RT MasterMix and 6 μL Nuclease-free H 2 After brief centrifugation in a handheld centrifuge, cDNA was synthesized according to the following program: 25°C, 10 min; 42°C, 50 min; 85°C, 5 min.

[0028] 1 μL cDNA, 0.2 μL upstream primer, 0.2 μL downstream primer, 10 μL SYBR ® Premix Ex Taq ™ Mix with 8.6 μL sterile water in an eight-tube tube. Set the program as pre-denaturation at 95 °C for 180 s, amplification at 95 °C for 30 s, 60 °C for 30 s, 72 °C for 30 s, and 40 cycles. PCR amplification was performed using GAPDH as an internal reference.

[0029] The mRNA expression level of the blank control group was taken as 1, and the ratio of the mRNA expression level of each intervention group to that of the blank control group was taken as the final result.

[0030] As shown in Table 1 , the screened raw materials GABA (purchased from Tianjin Century Weikang Biotechnology Co., Ltd.), tea theanine (purchased from Tianjin Century Weikang Biotechnology Co., Ltd.), Poria cocos extract (purchased from Shaanxi Ducheng Pharmaceutical Technology Co., Ltd.), lily extract (purchased from Xi'an Fenghe Biotechnology Co., Ltd.), polygonatum extract (purchased from Shaanxi Yunhe Biotechnology Co., Ltd.) and phosphatidylserine (purchased from Xi'an Muyuan Biotechnology Co., Ltd.) all have the activity of upregulating the relative expression of GABA mRNA and 5-HT mRNA.

[0031] When formulating food, it is necessary to comprehensively evaluate multiple dimensions such as biologically active functions, economic costs, sensory characteristics and safety. Specific requirements include: the interaction between the nutrients and functional ingredients of raw materials, price stability and seasonal fluctuations, and their storage stability; at the same time, attention should also be paid to sensory characteristics such as texture, taste, aroma, appearance and color; in terms of safety, it is necessary to evaluate potential allergic risks, the ability to prevent diseases, etc. In addition, factors such as the sustainability, eco-friendliness and social responsibility of raw materials, recommended intake and processing methods should also be considered. By comprehensively considering the above factors, a high-quality and safe food formula raw material selection criteria that meet the standards are formulated, and then GABA and Poria cocos extract are selected as basic raw materials in the formula, and Polygonatum sibiricum extract and phosphatidylserine are selected as further additives for subsequent research on formulas and specific additive amounts.

[0032] Table 1 Effects of formula ingredients on the relative expression of GABA mRNA and 5-HT mRNA

[0033] Example 2 Effect of the composition on the relative expression of GABA mRNA and 5-HT mRNA After weighing the raw materials according to the following formula, the raw materials were mixed in a double-screw cone mixer for 15-30 minutes to obtain a composition.

[0034] The raw materials of composition I include, by weight: 85 g of GABA, 15 g of Poria cocos extract, 40 g of Polygonatum sibiricum extract, and 5 g of phosphatidylserine.

[0035] The raw materials of composition II include, by weight: 70 g of GABA, 30 g of Poria cocos extract, 40 g of Polygonatum sibiricum extract, and 5 g of phosphatidylserine.

[0036] The raw materials of composition III include, by weight: 55 g of GABA, 45 g of Poria cocos extract, 40 g of Polygonatum sibiricum extract, and 5 g of phosphatidylserine.

[0037] The raw materials of composition IV include, by weight: 66 g of GABA, 34 g of Poria cocos extract, 40 g of Polygonatum sibiricum extract, and 5 g of phosphatidylserine.

[0038] The raw materials of composition V include, by weight: 77 g of GABA, 23 g of Poria cocos extract, 40 g of Polygonatum sibiricum extract, and 5 g of phosphatidylserine.

[0039] The raw materials of composition VI include, by weight: 55 g of GABA, 45 g of Poria cocos extract, 30 g of Polygonatum sibiricum extract, and 15 g of phosphatidylserine.

[0040] The raw materials of composition VII include, by weight: 55 g of GABA, 45 g of Poria cocos extract, 43 g of Polygonatum sibiricum extract, and 2 g of phosphatidylserine.

[0041] The raw materials of composition VIII include, by weight: 79.75 g of GABA and 65.25 g of Poria cocos extract.

[0042] The raw materials of Comparative Example 1 include, by weight: 145 g of Poria cocos extract.

[0043] Comparative Example 2 The raw materials include, by weight: GABA 145 g.

[0044] The raw materials of Comparative Example 3 include, by weight: 145 g of Polygonatum sibiricum extract.

[0045] The raw materials of Comparative Example 4 include, by weight: 145 g of phosphatidylserine.

[0046] The method in Example 1 was used to determine the effects of Comparative Examples 1-4 and Compositions I-VII on the relative expression of GABA mRNA and 5-HT mRNA to characterize the potential sleep-improving activity of the composition, and the results are shown in Table 2. The results show that the composition formula after adding GABA and Poria extract greatly improves the up-regulation activity of GABA mRNA and 5-HT mRNA, which is much higher than the activity when GABA and Poria extract are present separately. When GABA and Poria extract are present at the same time, the relative expression of GABA mRNA and 5-HT mRNA reaches 16.897±1.123 and 22.150±0.710, respectively. It can be seen that GABA and Poria extract synergistically play a role in improving sleep. Although the up-regulation activity of GABA mRNA and 5-HT mRNA will increase after increasing the dosage of the composition, the comprehensive sensory evaluation results finally determine that composition III has the best potential sleep-improving activity, and the ratio of GABA, Poria extract, Polygonatum sibiricum extract and phosphatidylserine is 11:9:8:1.

[0047] Table 2 Effects of the composition on the relative expression of GABA mRNA and 5-HT mRNA

[0048] Example 3. Animal experiments prove the sleep-improving effect of the composition 70 ICR male mice were selected and raised at 23±3 ℃; 65±5% humidity, 12 h light-dark cycle, and good ventilation. Before the experiment, the mice were raised for 1 week to adapt to the laboratory environment. The mice were randomly divided into blank control group, model group, low-dose formula group, medium-dose formula group, high-dose formula group, comparative example group 1, comparative example group 2, comparative example group 3, comparative example group 4, and positive drug group (10 mice in each group). Among them, the blank control group was injected with normal saline; except for the blank control group, the remaining groups were intraperitoneally injected with para-chlorophenylalanine (PACA) (450 mg / kg·bw) for 3 days to establish an insomnia model.

[0049] After the modeling period, the blank control group and the model group were gavaged with normal saline, the low-dose group was gavaged with low-dose composition III (76 mg / kg·bw), the medium-dose group was gavaged with medium-dose composition III (95 mg / kg·bw), the high-dose group was gavaged with high-dose composition III (114 mg / kg·bw), the comparative example 1 group was gavaged with Poria cocos extract (95 mg / kg·bw), the comparative example 2 group was gavaged with GABA (95 mg / kg·bw), the comparative example 3 group was gavaged with Polygonatum sibiricum extract (95 mg / kg·bw), the comparative example 4 group was gavaged with phosphatidylserine (95 mg / kg·bw), and the positive drug group was gavaged with estazolam solution (2 mg / kg·bw).

[0050] After 30 days of administration, 30 min after gavage, sodium pentobarbital (45 mg / kg·bw) was injected into each group of mice, and the righting reflex of the mice was observed. The mice were placed in a dorsal recumbency position. If the abdomen remained upward for 30 to 60 s, the righting reflex was lost. If the mice successfully turned over more than 3 times within 30 s, the righting reflex was recovered. The time from injection to the disappearance of the righting reflex was recorded as the sleep latency; the time from the disappearance of the righting reflex to the recovery of the righting reflex was the sleep duration.

[0051] like Figure 1 Compared with the blank control group, the sleep latency of the model group increased significantly, indicating that the model was successfully established. Compared with the model group, the sleep latency induced by sodium pentobarbital in the positive drug group, low-dose formula group, medium-dose formula group, and low-dose formula group was significantly shortened, which were (93±15) s, (117±11) s, (106±18) s, and (104±14) s, respectively. The formula group shortened the sleep latency of mice by 19.15%, 24.82%, and 26.24%, respectively. Among them, the medium-dose formula group and the high-dose formula group had the best effect on shortening the sleep latency of mice.

[0052] The length of sleep time is directly related to sleep quality. Figure 2Compared with the blank control group, the sleep time of the model group decreased significantly, indicating that the model was successfully established. Compared with the model group, the sleep time induced by sodium pentobarbital in the positive drug group, the low-dose group, the medium-dose group, and the low-dose group was significantly prolonged, which were (4055±252) s, (2283±434) s, (2900±340) s, and (2784±225) s, respectively. Among them, the medium-dose and high-dose groups had the best effect on prolonging the sleep time of mice. Compared with the medium-dose group, the effect of comparative examples 1-4 on prolonging the sleep time of mice was significantly weakened, which were (2107±206) s, (2354±169) s, (1875±126) s, and (1563±117) s, respectively, indicating that the intervention effect of composition III was better than that of each single component at the same dose, that is, there was a synergistic effect of improving sleep between the substances.

[0053] The ELISA kit was used to detect the GABA content in serum. Figure 3 As shown in (A), the GABA content in the serum of the model group (1.202±0.092 µmoL / L) was significantly lower (P<0.05) than that of the control group (3.404±0.078 µmoL / L), indicating that PACA modeling was successful and the secretion of inhibitory neurotransmitter GABA in ICR mice was inhibited. Compared with the model group, the serum GABA content in the low-dose group, medium-dose group, high-dose group, and positive drug group was significantly increased (P<0.05), which were (2.761±0.219) µmoL / L, (3.358±0.096) µmoL / L, (3.028±0.085) µmoL / L, and (3.279±0.104) µmoL / L, respectively. The GABA content in the mouse brain is shown in Figure 3 As shown in (B), the brain GABA content in the model group (2.830±0.153 µmoL / L) was significantly lower than that in the control group (4.502±0.122 µmoL / L). Compared with the model group, the brain GABA content in the low-dose group, medium-dose group, high-dose group, and positive drug group was significantly increased (P<0.05), which were (3.233±0.087) µmoL / L, (4.060±0.070) µmoL / L, (3.826±0.067) µmoL / L, and (4.094±0.088) µmoL / L, respectively. This indicates that the formula intervention can significantly inhibit the decrease in GABA secretion levels caused by insomnia, among which the medium-dose group has the best effect in increasing the serum GABA content.

[0054] The 5-HT concentration in mouse serum was detected by ELISA kit. Figure 3(C). Compared with the control group, the 5-HT concentration in the mouse serum (108.942±6.911) pg / mL decreased significantly after PCPA modeling (P<0.05), indicating that PCPA has an inhibitory effect on the synthesis of 5-HT in mice. Compared with the model group, the 5-HT content in the serum of the low-dose group, medium-dose group, high-dose group, and positive drug group was significantly increased (P<0.05), which were (131.378±5.524) pg / mL, (145.321±8.456) pg / mL, (140.673±8.526) pg / mL, and (150.687±4.137) pg / mL, respectively. The 5-HT content in the mouse brain is shown in Figure 3 As shown in (D), the 5-HT content in the brain of the model group mice (95.577±10.533 pg / mL) was significantly lower than that in the control group (203.750±13.358 pg / mL). Compared with the model group, the 5-HT content in the serum of the low-dose group, medium-dose group, high-dose group, and positive drug group was significantly increased (P<0.05), which were (139.327±6.731) pg / mL, (168.173±5.742) pg / mL, (176.827±6.911) pg / mL, and (225.865±7.752) pg / mL, respectively. This indicates that the formula treatment can significantly improve the decrease in 5-HT levels caused by insomnia, among which the medium and high doses have the best effects.

[0055] Example 4 Effect of the composition on relieving stress in mice Adrenocorticotropic hormone (ACTH) is a polypeptide hormone secreted by the pituitary gland of vertebrates. It can promote tissue proliferation of the adrenal cortex and the production and secretion of cortical hormones. When the body is sleep deprived and in a high-stress state, the amount of ACTH synthesis will increase. The ELISA kit was used to detect the ACTH concentration in mouse serum. The results are as follows Figure 4 (A) As shown. Compared with the control group, the serum ACTH concentration (86.752±2.229) ng / L of mice was significantly increased after PCPA modeling (P<0.05), indicating that PCPA increased the stress level of mice. Compared with the model group, the serum ACTH content of the low-dose group, medium-dose group, high-dose group, and positive drug group was significantly decreased (P<0.05), which were (64.58±3.95) ng / L, (61.90±1.89) ng / L, (72.49±3.57) ng / L, and (64.39±1.53) ng / L, respectively. This indicates that the formula can significantly improve the increase in ACTH content caused by insomnia, among which the medium-dose group has the best improvement effect.

[0056] Corticosterone (CORT) is a steroid hormone produced in the adrenal cortex that binds to glucocorticoid and mineralocorticoid receptors. It is produced in response to ACTH and is a precursor for aldosterone synthesis. Since glucocorticoid production increases due to stress, it is often used as a biomarker of stress. The concentration of CORT in mouse serum was measured using an ELISA kit. The results are as follows Figure 4 (B) Compared with the control group, the CORT concentration in the serum of mice (56.397±3.524) µg / L increased significantly after PCPA modeling (P<0.05), indicating that PCPA increased the stress level of mice. Compared with the model group, the serum CORT content of the low-dose group, medium-dose group, high-dose group, and positive drug group decreased significantly (P<0.05), which were (47.73±3.34) µg / L, (40.62±2.54) µg / L, (43.9±3.45) µg / L, and (42.35±1.20) µg / L, respectively. This indicates that the formula can significantly improve the increase in CORT content caused by insomnia, among which the medium-dose group has the best improvement effect.

[0057] Oxidative stress refers to a state of imbalance between oxidation and antioxidant effects in the body, which leads to the accumulation of reactive oxygen species (ROS) in the body and oxidative damage to tissues and organs. Oxidative stress is a negative effect produced by free radicals in the body and is considered to be an important factor leading to aging and disease. In the antioxidant protection system, superoxide dismutase (SOD) and catalase (CAT) are the main antioxidant enzymes. 2 O 2 ), which can effectively offset the harmful effects of ROS, assist in scavenging free radicals, and reduce and eliminate oxidative damage.

[0058] The ELISA kit was used to detect the SOD concentration in mouse serum. Figure 5 As shown in (A). Compared with the control group, the SOD concentration in the serum of mice (2.855±0.221) U / mL decreased significantly after PCPA modeling (P<0.05), indicating that insomnia caused oxidative damage in the mice. Compared with the model group, the SOD content in the serum of the low-dose group, medium-dose group, high-dose group, and positive drug group increased significantly (P<0.05), which were (5.415±0.304) U / mL, (6.238±0.228) U / mL, (5.706±0.191) U / mL, and (7.602±0.312) U / mL, respectively. The CAT content in the serum of mice is shown in Figure 5As shown in (B). Compared with the control group, the CAT concentration in the mouse serum (6.724±0.221) µmoL / mL decreased significantly after PCPA modeling (P<0.05). Compared with the model group, the CAT content in the serum of the low-dose group, medium-dose group, high-dose group, and positive drug group increased significantly (P<0.05), which were (9.992±1.043) µmoL / mL, (13.001±1.228) µmoL / mL, (11.986±0.624) µmoL / mL, and (11.833±0.781) µmoL / mL, respectively. The GSH content in mouse serum is shown in Figure 5 (C) Compared with the control group, the concentration of GSH in the serum of mice (3.587±0.856) µmoL / L decreased significantly after PCPA modeling (P<0.05). Compared with the model group, the serum CAT content of the low-dose group, medium-dose group, high-dose group, and positive drug group increased significantly (P<0.05), which were (13.659±2.081) µmoL / L, (15.438±1.266) µmoL / L, (15.323±1.263) µmoL / L, and (21.980±0.820) µmoL / L, respectively. In summary, the formula group can significantly reduce the oxidative damage caused by insomnia, among which the medium-dose group has the best effect.

[0059] Lack of sleep can also increase the risk of inflammation in the body, leading to adverse reactions such as cardiovascular disease, hypertension, and diabetes. In order to evaluate the impact of changes in sleep quality on the level of inflammation in the body, the levels of several inflammatory factors in mouse serum were detected. Figure 6 As shown in (A). Compared with the control group, the TNF-α concentration in the mouse serum (601.250±14.387) pg / mL increased significantly after PCPA modeling (P<0.05), indicating that insomnia increased the content of pro-inflammatory factors in mice. Compared with the model group, the TNF-α content in the serum of the low-dose group, medium-dose group, high-dose group, and positive drug group decreased significantly (P<0.05), which were (519.487±13.483) pg / mL, (503.170±12.830) pg / mL, (489.100±15.927) pg / mL, and (459.286±15.290) pg / mL, respectively. The IL-1β content in mouse serum is shown in Figure 6As shown in (B). Compared with the control group, the IL-1β concentration in the mouse serum (64.905±1.597) pg / mL increased significantly after PCPA modeling (P<0.05). Compared with the model group, the IL-1β content in the serum of the low-dose group, medium-dose group, high-dose group, and positive drug group decreased significantly (P<0.05), which were (61.237±0.673) pg / mL, (56.328±1.977) pg / mL, (54.643±1.275) pg / mL, and (53.226±1.779) pg / mL, respectively. The IL-6 content in the mouse serum is shown in Figure 6 (C) As shown. Compared with the control group, the IL-6 concentration in the mouse serum (108.327±5.297) pg / mL increased significantly after PCPA modeling (P<0.05). Compared with the model group, the IL-6 content in the serum of the low-dose group, medium-dose group, high-dose group, and positive drug group decreased significantly (P<0.05), which were (88.329±4.936) pg / mL, (82.033±3.275) pg / mL, (93.927±1.328) pg / mL, and (77.832±4.972) pg / mL, respectively. In summary, the formula group can significantly alleviate the increased inflammatory level caused by insomnia, among which the medium-dose group has the best effect.

[0060] HE staining was used to analyze the effect of formula intervention on hippocampal neurons in mice. Figure 7 As shown. In the blank control group, there were abundant neuronal cells in the hippocampus, with plump and round cell morphology, light staining, and neat arrangement. In the insomnia model group, neuronal cells were disorderly arranged, the cells were degenerated and atrophied, the shape was irregular, dark staining appeared, and the nuclei were condensed. After positive drug intervention, the cell morphology returned to normal. In the goat milk group, only a few cells showed normal morphology, most cells were still atrophied, and the number of cells was small. After intervention by the formula group, the damage to neuronal cells was repaired to varying degrees. Among them, the medium-dose and high-dose groups showed that the proportion of normal cells was higher, and most of the cells were neatly and tightly arranged, plump in morphology, and lightly stained.

[0061] c-Fos protein is a nuclear phosphoprotein, a product of immediate early genes (IEG), involved in regulating the excitability and survival of neurons. The expression of c-Fos is closely related to stress response. After stress stimulation, it has been confirmed that the expression of c-Fos protein increases in the hypothalamus, hippocampus and other areas related to cognition and anxiety-like behavior. By using c-Fos antibody to perform specific immunofluorescence staining on mouse hippocampal neurons to compare the stress levels of different treatment groups, the results are as follows: Figure 8The results showed that compared with the blank control group, a large number of c-Fos+ neurons appeared in the insomnia model group, indicating that insomnia induced neuronal activation and upregulated the body's stress response. After intervention in the formula group, c-Fos in the hippocampus of mice + The neuron density decreased significantly, indicating that the formula intervention effectively reversed the stress caused by insomnia. The c-Fos + The neuron density is the lowest.

[0062] The present invention proves that the composition can upregulate the expression levels of GABAAR mRNA and 5-HT1AR mRNA in SH-SY5Y neuronal cells by RT-PCR method. Further, through the sodium pentobarbital induced sleep experiment, it is proved that the composition can shorten the sleep latency and prolong the sleep time. At the same time, the GABA and 5-hydroxytryptamine (5-HT) contents in serum and brain tissue are detected, which proves that the composition can promote the release of neurotransmitters related to sleep improvement. In addition, by analyzing the levels of adrenocorticotropic hormone corticotropin (ACTH), corticosterone (CORT), superoxide dismutase (SOD), catalase (CAT), and reduced glutathione (GSH) in serum, it is proved that the composition has the effect of alleviating oxidative stress damage in the body; by detecting interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor (TNF-α) in serum, it is found that the composition can effectively downregulate the secretion of proinflammatory factors. In addition, the composition can inhibit the expression of c-Fos protein related to neuronal stress in the hippocampus and alleviate the degeneration and atrophy of hippocampal neuronal cells caused by insomnia.

[0063] The embodiments described above are only some embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work shall all fall within the scope of protection of the present invention.

Claims

1. A composition for improving sleep, characterized in that: The composition comprises gamma-aminobutyric acid and Poria cocos extract, and the mass ratio of the gamma-aminobutyric acid to the Poria cocos extract is 55-85:15-45.

2. The composition according to claim 1, characterized in that The mass ratio of the γ-aminobutyric acid to the Poria cocos extract is 11:

9.

3. The composition according to claim 1, characterized in that The composition further comprises a polygonatum extract and phosphatidylserine, and the weight ratio of the gamma-aminobutyric acid, the tuckahoe extract, the polygonatum extract and the phosphatidylserine is 55-85:15-45:30-40:2-15.

4. The composition according to claim 3, characterized in that The weight ratio of the gamma-aminobutyric acid, the Poria cocos extract, the Polygonatum sibiricum extract and the phosphatidylserine is 11:9:8:

1.

5. A product, characterized in that The product comprises the composition according to any one of claims 1 to 4.

6. The product according to claim 5, characterized in that The product is a health food or a medicine, and the product also includes excipients acceptable to the health food or medicine.

7. Use of the composition according to any one of claims 1 to 4 in preparing a product having a sleep-improving effect, characterized in that: The product is a health food or medicine.

8. The use according to claim 7, characterized in that: The application is application in preparing products capable of prolonging sleep time.

9. The use according to claim 7, characterized in that: The application is one or more of the following: (1) Application in the preparation of products capable of improving the decreased GABA secretion level caused by insomnia; (2) Application in the preparation of products capable of improving the decrease in 5-HT levels caused by insomnia; (3) Application in the preparation of products capable of improving the increase of ACTH content caused by insomnia; (4) Application in the preparation of products capable of improving the increase in CORT content caused by insomnia; (5) Application in the preparation of products capable of improving oxidative damage caused by insomnia; (6) Use in the preparation of products capable of improving increased inflammation levels caused by insomnia; (7) Use in the preparation of products capable of improving neuronal cell damage caused by insomnia.

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