Probiotics-tcm compound composition with sleep aid efficacy and preparation method and application thereof

Abstract

The application discloses a probiotic-TCM compound composition with sleep-aiding efficacy, which comprises a TCM compound and probiotics; wherein the TCM compound comprises a Spondias extract, a Poria cocos extract, a lily extract and a gardenia extract; and the probiotics are lactobacillus plantarum. The application further discloses a preparation method and application of the probiotic-TCM compound composition. The probiotic-TCM compound composition provided by the application can exert synergistic sleep-aiding efficacy, and is expected to become a safe and effective natural sleep-aiding solution, thereby providing a new approach for improving sleep quality.

Description

Technical Field

[0001] This invention relates to the field of health food technology, specifically to a probiotic-TCM compound composition with sleep-aiding effects, its preparation method, and its application. Background Technology

[0002] In modern society, sleep problems are becoming increasingly common, mainly manifested as insomnia, poor sleep quality, and insufficient sleep duration. According to China's "2021 White Paper on Exercise and Sleep," 300 million people in my country currently suffer from sleep disorders, and this number continues to rise annually. Insufficient sleep not only affects an individual's daily life and work efficiency but also leads to various health problems, such as cardiovascular disease, metabolic disorders, and mental illnesses.

[0003] Current solutions for improving sleep primarily rely on medication, including barbiturates, melatonin, benzodiazepines, and non-benzodiazepine drugs. While medication-assisted sleep is fast-acting and effective, it can cause varying degrees of side effects, including excessive daytime sleepiness. Long-term use can also lead to poor drug tolerance, cognitive impairment, dependence, and withdrawal symptoms. For example, melatonin, an indole hormone secreted by the pineal gland, regulates the sleep-wake cycle; however, long-term overuse can reduce melatonin secretion, causing insomnia. Benzodiazepines enhance GABA activity by acting on GABA receptors, thereby slowing neurotransmission and providing sedative and anti-anxiety effects. However, high doses or continuous use can lead to adverse reactions, including altered sleep structure, impaired daytime psychomotor performance, anterograde amnesia, rebound insomnia, and withdrawal symptoms. Non-benzodiazepine drugs have a similar hypnotic effect to benzodiazepines, but long-term use may lead to adverse reactions such as drug tolerance and addiction.

[0004] Therefore, finding a safe and effective sleep aid solution is an urgent problem to be solved. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a probiotic-TCM compound composition with sleep-aiding effects. The probiotics and TCM compound in the composition can exert a synergistic sleep-aiding effect, and are expected to become a safe and effective natural substance sleep-aiding solution, thus providing a new way to improve sleep quality.

[0006] To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0007] The first aspect of the present invention provides a probiotic-TCM compound composition with sleep-aiding effects, comprising a TCM compound and probiotics;

[0008] The TCM compound includes jujube seed extract, poria cocos extract, lily extract and gardenia extract.

[0009] The probiotic is Lactobacillus plantarum.

[0010] Traditional Chinese medicine's mechanisms for improving sleep mainly include regulating gut microbiota, anti-inflammation, and regulating neurotransmitters. For example, jujube seed, as a sedative ingredient, has various pharmacological effects, mainly including sedative-hypnotic and anti-anxiety bioactivities. Jujube seed is rich in saponins and flavonoids, which have significant sedative and hypnotic effects on mice, increasing the sleep time induced by sodium barbital. Poria cocos, as a diuretic ingredient, has triterpenes and polysaccharides as its effective components for sedation and hypnosis. Poria cocos water extract has a significant inhibitory effect on mouse movement, reducing sleep latency and prolonging sleep time. Lily bulb, as a calming and tranquilizing ingredient, has saponins as its effective component for sedation and tranquility. Lily bulb extract can significantly shorten the time to fall asleep in mice and, in conjunction with sodium pentobarbital, prolong the sleep time in mice. Gardenia jasminoides, as a heat-clearing ingredient, has iridoid ethers as its main sedative and tranquilizing substances. With prolonged exposure to lily bulb extract in rats, anxiety behavior in rats significantly decreased.

[0011] Probiotics such as Lactobacillus primarily improve sleep through neuroendocrine, vagus nerve, and immunomodulatory pathways. In the neuroendocrine pathway, probiotics mainly metabolize and produce the inhibitory neurotransmitter gamma-aminobutyric acid (GABA), which plays an important physiological role in the human body, exhibiting sedative, calming, and heart rate-regulating effects. In the vagus nerve pathway, probiotics can mediate host brain and behavior by activating vagus nerve signals. After vagotomy in mice, the previously effective probiotics could not reverse anxiety behavior. In the immunomodulatory pathway, probiotics can induce the secondary host to produce sleep-regulating cytokines, resulting in reduced REM sleep time and increased non-REM sleep time in mice.

[0012] The inventors discovered in their research that the TCM compound, composed of jujube seed extract, gardenia extract, lily extract, and poria cocos extract, when used in combination with the probiotic Lactobacillus plantarum in a certain proportion, can produce a synergistic effect in promoting sleep. It can prolong sleep time, shorten sleep latency, increase the incidence of sleep, and thus improve sleep quality. Furthermore, this composition is highly effective in promoting sleep and has a high safety profile with no side effects.

[0013] Furthermore, by weight, the probiotic-TCM compound composition with sleep-aiding effects comprises: 3-8 parts of probiotics and 2-5 parts of TCM compound. The weight portions of probiotics can be 3, 4, 5, 6, 7, or 8 parts, etc.; the weight portions of TCM compound can be 2, 3, 4, or 5 parts, etc.

[0014] In a preferred embodiment of the present invention, the ratio of probiotics to TCM compound in the composition is (4-6):(3-5).

[0015] Furthermore, the probiotic is Lactobacillus plantarum powder, and the bacterial content in the powder can be 1*102 10 CFU / g.

[0016] Furthermore, the TCM compound comprises the following components by weight: 15-30 parts of Ziziphus jujuba seed extract, 0-4 parts of Poria cocos extract, 0-12 parts of Lilium brownii extract, and 5-12 parts of Gardenia jasminoides extract.

[0017] In the TCM compound of the present invention, the weight parts of jujube seed extract are 15 to 30 parts, for example, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 parts, etc.; the weight parts of Poria cocos extract are 0 to 4 parts, for example, 0, 1, 2, 3, 4 parts, etc.; the weight parts of lily extract are 0 to 12 parts, for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 parts, etc.; and the weight parts of gardenia extract are 5 to 12 parts, for example, 5, 6, 7, 8, 9, 10, 11, 12 parts, etc.

[0018] In a preferred embodiment of the present invention, the mass ratio of Ziziphus jujuba seed extract, Poria cocos extract, Lilium brownii extract and Gardenia jasminoides extract in the TCM compound is 8:1:3:3.

[0019] In another preferred embodiment of the present invention, the TCM compound includes jujube seed extract and gardenia extract, and the mass ratio of jujube seed extract to gardenia extract is 8:3.

[0020] A second aspect of this invention provides a method for preparing a probiotic-TCM compound composition with sleep-aiding effects, comprising the following steps:

[0021] S1. The raw material for the extract is extracted twice with water at a temperature of 90-100℃, and then impurities are filtered out to obtain the extract; the raw material for the extract is jujube seed powder, poria cocos powder, lily powder or gardenia powder;

[0022] S2. The extract is concentrated under vacuum to a relative density of 1.09–1.12 g / cm³. 3 Then add maltodextrin and autoclave.

[0023] S3. In a clean environment, the sterilized extract is spray-dried to obtain a powdered material; after crushing, sieving, and mixing, the extract powder is obtained.

[0024] S4. Mix the extract powder with the probiotic powder to obtain the probiotic-TCM compound composition.

[0025] Further, in step S1, when the raw material for the extract is jujube seed powder, the jujube seed powder is soaked in 8 times and 6 times the amount of water, respectively, and the extraction time is 120 minutes and 60 minutes, respectively.

[0026] When the raw material for the extract is Poria cocos powder, the Poria cocos powder is soaked in 10 times and 8 times the amount of water, and the extraction time is 120 minutes and 60 minutes, respectively.

[0027] When the raw material for the extract is lily powder, the lily powder is soaked in 8 times and 8 times the amount of water, respectively, and the extraction time is 120 minutes and 60 minutes, respectively.

[0028] When the raw material for the extract is gardenia powder, the gardenia powder is soaked in 8 times and 6 times the amount of water, respectively, and the extraction time is 120 minutes and 60 minutes, respectively.

[0029] Furthermore, in step S2, the vacuum degree is controlled at -0.06 to -0.09 MPa and the concentration temperature is 50-75℃ during the concentration process; the temperature is controlled at 115-135℃ and the time is 15-30 minutes during the autoclaving process.

[0030] Furthermore, in step S3, during spray drying, the inlet air temperature is 180-205℃ and the outlet air temperature is 85-105℃.

[0031] Furthermore, taking jujube seed extract as an example, its extraction method is as follows:

[0032] a1 Raw material source and preliminary testing: After receiving the jujube seed powder raw material, it undergoes strict testing by the quality control (QC) department to ensure that it meets the standards. The raw materials that pass the inspection enter the extraction process.

[0033] a2 Extraction and Filtration: Water extraction twice: Jujube seed powder was extracted with 8 times and 6 times the amount of water, respectively. The first extraction time was 120 minutes, and the second time was 60 minutes. The temperature was maintained at 90℃ to 100℃ during the extraction process, and continuous stirring was carried out to ensure the full extraction of active ingredients; Filtration: After each extraction, the extract was filtered through a filter cloth or filter screen to remove insoluble impurities, ensuring that the extract was clear and contained high-purity active ingredients.

[0034] a3 Vacuum Concentration and Maltodextrin Addition: Concentration Conditions: The filtered extract is processed using a vacuum concentration device, with the vacuum level controlled at -0.06 to -0.09 MPa and the temperature set at 50℃ to 75℃, concentrated to a relative density of 1.09 to 1.12 to ensure the extract is within the optimal concentration range; Maltodextrin Addition: An appropriate amount of maltodextrin (according to the specified ratio) is added to the concentrated extract to enhance product stability and finished product characteristics. The concentrate is further heated and stirred evenly to ensure complete dissolution of the maltodextrin.

[0035] a4 High-Temperature Sterilization and Drying: High-Pressure Sterilization: The extract containing maltodextrin is sterilized at 115℃ to 135℃ for 15-30 minutes to ensure complete inactivation of microorganisms in the extract; Spray Drying: The sterilized extract is spray dried in a clean production area (Grade D) (inlet air temperature controlled at 180℃ to 205℃, outlet air temperature at 85℃ to 105℃) to obtain a powdered product. The moisture content of the powder is controlled within the specified range to ensure product quality.

[0036] a5 Crushing and Sieving: Crushing: The dried jujube seed powder needs to undergo a crushing process to ensure it reaches a uniform particle size; Sieving: The powder is initially sieved using a 100-mesh sieve to remove larger particles. It is then sieved again under CCP (Critical Control Point) standards to ensure consistent product particle size.

[0037] a6 Mixing and Packaging: Mixing: The sieved jujube seed powder is mixed evenly according to the ratio for 50 to 60 minutes to ensure the uniformity and stability of the powder; Inner Packaging Inspection: The mixed powder undergoes internal control inspection to confirm that all indicators are qualified. The jujube seed powder that passes the inspection is then sealed in the inner packaging, followed by outer packaging; Finished Product Inspection and Warehousing: The packaged finished product is inspected by a metal foreign object detector (CCP3). After passing the inspection, a label is affixed, indicating the product name, batch number, and net content, and the product is then stored in the warehouse for later use.

[0038] Further, in step S4, the various raw materials in the TCM compound are mixed according to the ratio, pulverized, and passed through a 150-mesh sieve; then the obtained TCM compound is thoroughly mixed with probiotic powder, and after passing the inspection, it is packed into a sterilized aluminum foil bag to obtain the probiotic-TCM compound composition.

[0039] A third aspect of the present invention provides the application of the aforementioned probiotic-TCM compound composition in the preparation of sleep aid formulations.

[0040] Furthermore, the sleep aid is used to increase the content of inhibitory neurotransmitters in the brain, decrease the content of excitatory neurotransmitters, and reduce the expression of GFAP protein.

[0041] Furthermore, the formulations include, but are not limited to, food, pharmaceuticals, and health products.

[0042] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0043] 1. The probiotic-TCM compound composition provided by this invention has no direct hypnotic effect, but it can prolong sleep time, shorten sleep latency, and increase the incidence of sleep, thus improving sleep. This probiotic-TCM compound composition improves sleep by increasing the content of inhibitory neurotransmitters in the mouse brain, decreasing the content of excitatory neurotransmitters, and reducing the expression of GFAP protein.

[0044] 2. The probiotic-TCM compound composition provided by this invention contains no preservatives or artificial synthetic chemical components. While having sleep-aiding effects, it is highly safe, has no toxic side effects, and is suitable for long-term use. Attached Figure Description

[0045] Figure 1 Figure showing the effect of probiotics combined with TCM compound on sleep in mice;

[0046] Figure 2 Figure showing the effect of probiotics combined with TCM compound on the sleep latency of sodium barbital in mice;

[0047] Figure 3 Figure showing the effect of probiotics combined with TCM compound on neurotransmitters in mouse brain tissue;

[0048] Figure 4 Figure showing the effect of probiotics combined with TCM compound on GFAP expression in mouse brain;

[0049] Among them: A: blank group; B: probiotic group; C: TCM compound 1 group; D: TCM compound 2 group; E: probiotic + TCM compound 1 group; F: probiotic + TCM compound 2 group; G: positive control group. Detailed Implementation

[0050] The present invention will be further described below with reference to specific embodiments, so that those skilled in the art can better understand and implement the present invention, but the embodiments are not intended to limit the present invention.

[0051] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0052] Unless otherwise specified, the experimental methods used in the following examples and comparative examples are conventional methods, and the materials and reagents used are commercially available unless otherwise specified.

[0053] Example 1

[0054] This embodiment provides a probiotic-TCM compound composition with sleep-aiding effects, comprising 3 parts TCM compound and 40 parts probiotics by weight.

[0055] The TCM compound comprises jujube seed extract, poria cocos extract, lily extract, and gardenia extract in a mass ratio of 8:1:3:3, designated as TCM compound group 1. The probiotic is Lactobacillus plantarum P8 powder.

[0056] Example 2

[0057] This embodiment provides a probiotic-TCM compound composition with sleep-aiding effects, comprising 3 parts TCM compound and 40 parts probiotics by weight.

[0058] The TCM compound comprises jujube seed extract and gardenia extract in a mass ratio of 8:3, designated as TCM compound group 2. The probiotic is Lactobacillus plantarum P-8 powder.

[0059] Comparative Example 1

[0060] This embodiment provides a TCM compound composition with sleep-aiding effects, comprising jujube seed extract, poria cocos extract, lily extract and gardenia extract, in a mass ratio of 8:1:3:3, referred to as TCM compound group 1.

[0061] Comparative Example 2

[0062] This embodiment provides a TCM compound composition with sleep-aiding effects, comprising jujube seed extract and gardenia extract in a mass ratio of 8:3, referred to as TCM compound group 2.

[0063] Comparative Example 3

[0064] This embodiment provides a probiotic powder, which is Lactobacillus plantarum P-8 powder.

[0065] Sleep aid efficacy test

[0066] 1. Experimental Methods

[0067] 1.1. Experimental animals: Eighty-four-week-old ICR mice were housed in an SPF (pathogen-free) laboratory at a temperature of 22-24°C with 12-hour light-dark cycles, relative humidity of 40%-55%, and good ventilation. The animals had free access to food and water and were fed for one week before the experiments were conducted. Subsequent experiments continued for another 30 days.

[0068] 1.2. Animal grouping: 84 mice were randomly divided into 7 groups of 12 mice each: blank group (normal feeding group), probiotic group, TCM group (2 groups), probiotic + TCM group (2 groups), and positive control group.

[0069] 1.3. Animal administration method:

[0070] All mice in all groups were fed normally and administered medication as directed:

[0071] (1) The blank group received 0.1 mL of physiological saline by gavage;

[0072] (2) The probiotic group was given 11 mg / day / pcs of probiotic dry powder as a daily dose by gavage to the control group;

[0073] (3) TCM1 group was given TCM compound of Comparative Example 1 in the feed every day (TCM compound added amount was 0.38g / 100g - equivalent to 11mg / d / pcs per bird);

[0074] (4) The TCM2 group was given a fixed amount of TCM compound from Comparative Example 2 in the feed every day (TCM compound addition amount was 0.38g / 100g - equivalent to 11mg / d / pcs per animal);

[0075] (5) The probiotic + TCM1 group was given a quantitative amount of TCM compound from Example 1 in the feed every day, and a quantitative amount of probiotic dry powder was administered by gavage (TCM compound added amount was 0.21g / 100g - equivalent to 6mg / d / pcs per animal, probiotic dry powder 5mg / d / pcs).

[0076] (6) The probiotic + TCM2 group was given a fixed amount of TCM compound from Example 2 in the feed every day, and a fixed amount of probiotic dry powder was administered by gavage (TCM compound added at 0.18g / 100g - equivalent to 5.2mg / d / pcs per animal, and probiotic dry powder at 5.8mg / d / pcs).

[0077] (7) The positive control group was treated with diazepam administered by gavage daily at a dose of 2 mg / kg / d.

[0078] 1.4. Behavioral Analysis

[0079] Preliminary treatments were conducted before the formal experiment to determine the sodium pentobarbital dose (40-60 mg / kg) that would induce 100% sleep in the animals without causing excessively long sleep duration. This dose was used in the formal experiment. Sleep patterns in each group of mice were observed after administration. If no immediate sleep was observed 30 minutes after the last gavage, a subthreshold dose of sodium pentobarbital was used for subsequent synergistic sleep behavior experiments (six mice were randomly assigned to each of the seven groups of 12) to avoid false positives.

[0080] 1.5. Physiological and Biochemical Analysis

[0081] Evaluation of mouse weight changes. Mice were euthanized (the remaining 6 mice from 7 groups were sacrificed) when there was no significant decrease in mouse weight and in accordance with ethical guidelines, and the organ-to-body weight ratio was analyzed.

[0082] 1.6. Neurotransmitter Analysis and Immunofluorescence Staining

[0083] Neurotransmitters were isolated and extracted from homogenized brain tissue of euthanized mice after buffer treatment, and the neurotransmitters 5-HT, DA and GABA were quantified by high performance liquid chromatography (HPLC).

[0084] Brain tissue cells were fixed with 4% paraformaldehyde for 10–20 minutes, and infiltrated cells were treated with 0.1% Triton X-100. Samples were incubated with anti-GFAP antibody, and GFAP was labeled with a secondary antibody conjugated with Alexa Fluor 488. Nuclear staining was performed using Hoechst 33342. Visualization and analysis were performed under a fluorescence microscope.

[0085] 2. Experimental Results

[0086] 2.1. Effects of Lactobacillus plantarum combined with TCM compound on mouse body weight

[0087] Mouse body weight reflects whether the mouse is growing healthily. Table 1 shows the effect of Lactobacillus plantarum combined with TCM compound on mouse body weight. As can be seen from the table, after 30 days of gavage, there was no significant difference in body weight among the groups (P > 0.05). This indicates that the probiotics combined with TCM compound have no side effects on mouse body weight and are safe.

[0088] Table 1

[0089] Blank group <![CDATA[34.22±1.97 a ]]> <![CDATA[36.98±2.75 a ]]> Probiotic group <![CDATA[33.23±1.80 a ]]> <![CDATA[36.12±2.17 a ]]> TCM1 Group <![CDATA[34.46±1.96 a ]]> <![CDATA[37.60±2.48 a ]]> TCM2 Group <![CDATA[34.96±2.15 a ]]> <![CDATA[36.98±2.50 a ]]> Probiotics + TCM1 group <![CDATA[33.98±1.79 a ]]> <![CDATA[35.70±2.35 a ]]> Probiotics + TCM2 group <![CDATA[33.61±1.85 a ]]> <![CDATA[35.38±3.63 a ]]> Positive control group <![CDATA[33.69±1.76 a ]]> <![CDATA[35.73±2.56 a ]]>

[0090] 2.2. Effects of Lactobacillus plantarum combined with TCM compound on organ coefficients in mice

[0091] As shown in Table 2, after 30 days of gavage, there were no significant differences in organ coefficients (such as lung, kidney, spleen, and liver) among the different groups of mice (P > 0.05). This indicates that the probiotics combined with TCM compound had no adverse effects on the organs of mice.

[0092] Table 2

[0093]

[0094] 2.3. Effects of Lactobacillus plantarum combined with TCM compound on direct sleep in mice

[0095] Studies have shown that if a sample has an inhibitory effect on the central nervous system, it will cause mice to fall asleep directly, indicating that the sample does not fall under the category of improving sleep. When no animals in the blank control group and the probiotic combined with TCM compound group fall asleep, it can be determined that the probiotic combined with TCM compound has no inhibitory effect on the central nervous system, and the remaining three behavioral experiments are then conducted.

[0096] As shown in Table 3, 30 minutes after the last gavage, neither the blank control group nor the probiotic combined with TCM compound group showed direct sleep, and there was no significant difference (P>0.05), indicating that the probiotic combined with TCM compound cannot induce direct sleep in mice.

[0097] Table 3

[0098] Blank group 12 <![CDATA[0 a ]]> Probiotic group 12 <![CDATA[0 a ]]> TCM1 Group 12 <![CDATA[0 a ]]> TCM2 Group 12 <![CDATA[0 a ]]> Probiotics + TCM1 group 12 <![CDATA[0 a ]]> Probiotics + TCM2 group 12 <![CDATA[0 a ]]> Positive control group 12 <![CDATA[0 a ]]>

[0099] 2.4. Effects of Lactobacillus plantarum combined with TCM compound on pentobarbital sodium-induced sleep time in mice

[0100] The length of sleep directly affects sleep quality. Sodium pentobarbital is a central nervous system drug with a hypnotic effect, which can effectively prolong sleep time. Compared with the blank control group, if the mice in the probiotic combined with TCM compound group had a significantly prolonged sleep time, it indicates that the sample can synergistically prolong the sleep time of mice with sodium pentobarbital.

[0101] like Figure 1 As shown, compared with the blank control group, the probiotic combined with TCM compound 2 group had a prolonged sleep time of 34.37 min. Therefore, the probiotic combined with TCM compound 2 group can enhance the effect of sodium pentobarbital, has the best sleep improvement effect, and can prolong the sleep time of mice.

[0102] 2.5. Effects of Lactobacillus plantarum combined with TCM compound on low-dose pentobarbital sodium-induced hypnosis in mice.

[0103] Sodium pentobarbital is a centrally acting drug that is metabolized by liver enzymes. If the sample inhibits liver enzymes, prolonged sleep time may also occur, leading to false positive results. To rule out false positives, a subthreshold dose of sodium pentobarbital for hypnosis should be performed. After the last administration, each mouse is intraperitoneally injected with sodium pentobarbital at a dose of 0.2 mL / 20 g (mouse), and the disappearance of the righting reflex is used as the indicator.

[0104] Table 4 shows that the incidence of sleep was increased in all groups of mice compared with the blank control group. Specifically, the incidence of sleep in the probiotic combined with TCM 1 group and the probiotic combined with TCM 2 group was 66.67% and 91.67%, respectively, both showing significant differences (P<0.05). These results indicate that probiotics, in combination with sodium pentobarbital, can synergistically increase the incidence of sleep.

[0105] Table 4

[0106] Blank group 12 2 <![CDATA[16.67% e ]]> Probiotic group 12 3 <![CDATA[25.00% d ]]> TCM1 Group 12 10 <![CDATA[83.33% b ]]> TCM2 Group 12 8 <![CDATA[66.67% c ]]> Probiotics + TCM1 group 12 8 <![CDATA[66.67% c ]]> Probiotics + TCM2 group 12 11 <![CDATA[91.67% a ]]> Positive control group 12 11 <![CDATA[91.67% a ]]>

[0107] 2.6. Effects of Lactobacillus plantarum combined with TCM compound on sodium barbital sleep latency in mice

[0108] The speed at which one falls asleep is also an important indicator of sleep quality; a short sleep latency indicates a rapid onset of sleep. Sodium barbital is a drug that has a depressant effect on the central nervous system. If, after intraperitoneal injection, the sleep latency in the probiotic combined with TCM compound group was significantly shortened, it indicates a synergistic effect between the sample and sodium barbital, and the experimental result is positive.

[0109] Depend on Figure 2 As shown, compared with the blank control group, the sleep time was shortened in both the probiotic combined with TCM compound group 1 and the probiotic combined with TCM compound group 2. Among them, the probiotic combined with TCM compound group 2 showed the best sleep improvement effect, with a sleep time shortened by 13 minutes. The results indicate that the probiotic combined with TCM compound has a synergistic effect with sodium barbital, which can shorten the sleep latency of mice.

[0110] 2.7. Effects of Lactobacillus plantarum combined with TCM compound on neurotransmitters in mouse brain tissue

[0111] Please see Figure 3 Compared with the blank control group and the positive control group, the levels of inhibitory neurotransmitters in the probiotic combined with TCM compound groups 1 and 2 were increased. Compared with the blank control group, the levels of excitatory neurotransmitters in the probiotic combined with TCM compound groups 1 and 2 were significantly decreased (P < 0.05). This indicates that the probiotic-TCM compound combination can effectively increase the levels of 5-HT and GABA in the mouse brain and decrease the level of DA, thereby assisting sleep by regulating the neurotransmitter balance mechanism.

[0112] 2.8. Effects of Lactobacillus plantarum combined with TCM compound on GFAP expression in mouse brain

[0113] In addition to neuronal circuits, the brain also contains various types of glial cells, which are 10-50 times more numerous than neurons. Among them, astrocytes (AS) are closely related to sleep. Studies have shown that astrocytes can maintain the sleep-wake cycle by releasing ATP from the brain. Glial fibrillary acidic protein (GFAP) is a type III intermediate filamentous protein that exists in monomeric form and is a marker of AS activation.

[0114] Depend on Figure 4 As can be seen, the green area represents GFAP. When sleep disorders occur, AS begins to activate, and GFAP expression increases. Compared with the control group, the GFAP expression levels in both probiotic combined with TCM compound groups 1 and 2 were downregulated, indicating that probiotic combined with TCM compound can assist sleep by regulating GFAP expression.

[0115] In summary, the probiotic-TCM compound composition had no effect on mouse body weight or organ coefficients, was non-toxic, and demonstrated safety. Based on epibehavioral results, the probiotic-TCM compound composition did not have a direct hypnotic effect, but it could prolong sleep time, shorten sleep latency, and increase the incidence of sleep, with all experimental results being positive, proving that the probiotic-TCM compound composition has a sleep-improving effect. Furthermore, based on brain tissue biochemical and histopathological analysis, it was found that the probiotic-TCM compound composition improves sleep by increasing the content of inhibitory neurotransmitters, decreasing the content of excitatory neurotransmitters, and reducing the expression of GFAP protein in the mouse brain. Therefore, the probiotic-TCM compound composition is effective, safe, and without side effects in aiding sleep.

[0116] The above-described embodiments are merely preferred embodiments provided to fully illustrate the present invention, and the scope of protection of the present invention is not limited thereto. Equivalent substitutions or modifications made by those skilled in the art based on the present invention are all within the scope of protection of the present invention. The scope of protection of the present invention is defined by the claims.

Claims

1. A probiotic-TCM compound composition with sleep-aiding effects, characterized in that, By weight, the composition comprises the following components: 3-8 parts probiotics and 2-5 parts TCM compound; The TCM compound is composed of jujube seed extract and gardenia extract, and the mass ratio of jujube seed extract to gardenia extract is 8:

3. The probiotic is Lactobacillus plantarum.

2. The probiotic-TCM compound composition with sleep-aiding effects according to claim 1, characterized in that, The probiotic is Lactobacillus plantarum powder, with a bacterial count of 1*10⁻⁶. 10 CFU / g.

3. A method for preparing a probiotic-TCM compound composition with sleep-aiding effects according to claim 1 or 2, characterized in that, Includes the following steps: S1. The raw material for the extract is extracted twice with water at a temperature of 90-100℃, and then impurities are filtered out to obtain the extract; the raw material for the extract is jujube seed powder or gardenia powder; S2. The extract is concentrated under vacuum to a relative density of 1.09~1.12 g / cm³. 3 Then add maltodextrin and autoclave. S3. In a clean environment, the sterilized extract is spray-dried to obtain a powder; the powder is then pulverized, sieved, and mixed to obtain jujube seed extract powder and gardenia extract powder, respectively. S4. The jujube seed extract powder and the gardenia extract powder are mixed together, and then mixed with probiotic powder to obtain the probiotic-TCM compound composition.

4. The method for preparing a probiotic-TCM compound composition with sleep-aiding effects according to claim 3, characterized in that, In step S1, the extract raw materials are soaked in 8 times and 6 times the amount of water, respectively, for 120 minutes and 60 minutes, respectively.

5. A method for preparing a probiotic-TCM compound composition with sleep-aiding effects according to claim 3, characterized in that, In step S2, the vacuum degree is controlled at -0.06 to -0.09 MPa and the concentration temperature is 50-75℃ during the concentration process; the temperature is controlled at 115-135℃ and the time is 15-30 minutes during the autoclaving process.

6. The method for preparing the probiotic-TCM compound composition with sleep-aiding effects according to claim 3, characterized in that, In step S3, during spray drying, the inlet air temperature is 180-205℃ and the outlet air temperature is 85-105℃.

7. The use of the probiotic-TCM compound composition according to claim 1 or 2 in the preparation of sleep aid formulations.

8. The application according to claim 7, characterized in that, The sleep aid preparation can increase the content of inhibitory neurotransmitters in the brain, decrease the content of excitatory neurotransmitters, and reduce the expression of GFAP protein.

9. The application according to claim 7, characterized in that, The preparations include pharmaceuticals and health products.

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