Probiotic-TCM compound composition with sleep aiding effect as well as preparation method and application of probiotic-TCM compound composition

Abstract

The invention discloses a probiotic-TCM compound composition with a sleep aiding effect. The probiotic-TCM compound composition comprises a TCM compound and probiotics, wherein the TCM compound comprises a spina date seed extract, a poria cocos extract, a lily bulb extract and a gardenia extract; the probiotics are lactobacillus plantarum. The invention further discloses a preparation method and application of the probiotic-TCM compound composition. According to the probiotic-TCM compound composition provided by the invention, the compound of the probiotics and the TCM can play a synergistic sleep-aiding role, and the probiotic-TCM compound composition is expected to become a safe and effective natural substance sleep-aiding solution, so that a new way is provided for improving the sleep quality.

Description

Technical Field

[0001] The present invention relates to the technical field of health food, and in particular to a probiotic-TCM compound composition with sleep-aiding effect, and a preparation method and application thereof. Background Art

[0002] In modern society, sleep problems are becoming increasingly common, mainly manifested as insomnia, poor sleep quality and insufficient sleep time. According to China's "2021 Sports and Sleep White Paper", there are currently 300 million people in my country with sleep disorders, and this number is still rising year by year. Lack of sleep not only affects a person's daily life and work efficiency, but also leads to a variety of health problems, such as cardiovascular disease, metabolic disorders, mental illness, etc.

[0003] The existing technical solutions for improving sleep are mainly based on drug treatment, including barbiturates, melatonin drugs, benzodiazepines, non-benzodiazepines, etc. Although drug-assisted sleep is fast and effective, it has different degrees of side effects on the human body, including excessive daytime sleepiness. At the same time, long-term use of people will produce poor tolerance to drugs, cognitive impairment, dependence and withdrawal reactions. For example, melatonin drugs are an indole hormone secreted by the pineal gland, which has the function of regulating the sleep-wake cycle, but long-term excessive use of melatonin drugs will reduce the secretion of melatonin in the body, thereby causing insomnia. Benzodiazepines enhance the activity of GABA by acting on GABA receptors, thereby slowing down neurotransmission and sedative and anti-anxiety effects. However, high doses or continuous use may lead to some adverse reactions, including changes in sleep structure, impaired daytime psychomotor performance, anterograde amnesia, rebound insomnia, withdrawal symptoms, etc. The hypnotic effects of non-benzodiazepines are similar to those of benzodiazepines, but if taken for a long time, adverse reactions such as tolerance and addiction may occur.

[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 effect. The probiotics and TCM compound in the composition can play a synergistic sleep-aiding effect, and is expected to become a safe and effective natural substance sleep-aiding solution, thereby providing a new way to improve sleep quality.

[0006] In order to solve the above technical problems, the present invention provides the following technical solutions:

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

[0008] The TCM compound comprises a Chinese date seed extract, a tuckahoe extract, a lily extract and a gardenia extract;

[0009] The probiotic is Lactobacillus plantarum.

[0010] The mechanisms of traditional Chinese medicine in improving sleep mainly include regulating intestinal flora, anti-inflammatory, regulating neurotransmitters and other aspects. For example, as a sedative raw material, Chinese date seeds have multiple pharmacological effects, mainly including sedation, hypnosis, anti-anxiety and other biological activities. Chinese date seeds are rich in saponins and flavonoids, which have significant sedative and hypnotic effects on mice and increase the sleep time induced by sodium pentobarbital. As a diuretic and dampness-inducing raw material, Poria cocos has triterpenes and polysaccharides as its active ingredients for sedative and hypnotic effects. Poria cocos water extract has a significant inhibitory effect on the movement of mice, can reduce sleep latency and prolong sleep time. Lily is a raw material for clearing the mind and calming the nerves, and its active ingredient for calming the nerves is saponin. Lily extract can significantly shorten the time for mice to fall asleep, and cooperate with sodium pentobarbital to prolong the sleep time of mice. Gardenia is a heat-clearing raw material, and its main substance for calming the nerves is cyclopentane physiologically active substances. As the exposure time of rats to lily extract substances increases, the anxiety behavior of rats is significantly reduced.

[0011] Probiotics such as Lactobacillus improve sleep mainly through neuroendocrine pathways, vagus nerve pathways, and immunomodulatory pathways. In the neuroendocrine pathway, probiotics mainly metabolize to produce inhibitory neurotransmitters γ-aminobutyric acid (GABA), which plays an important physiological role in the human body and has the effects of calming the nerves, regulating heart rate, etc. In the vagus nerve pathway, probiotics can mediate the host's brain and behavior by activating vagus nerve signals. After vagotomy in mice, the originally effective probiotics could not reverse anxious behavior. In the immunomodulatory pathway, probiotics can induce the secondary host to produce sleep regulation-related cytokines, resulting in less rapid eye movement sleep time and increased non-rapid eye movement sleep time in mice.

[0012] The inventors found in their research that the TCM compound composed of Semen Ziziphi Spinosae extract, Gardenia jasminoides extract, Lilium extract and Poria cocos extract, when used in combination with the probiotic Lactobacillus plantarum in a certain ratio, can produce a synergistic effect in helping sleep, prolonging sleep time, shortening sleep latency, increasing sleep incidence, and thus improving sleep. In addition, the composition has a high sleep-aiding effect, is highly safe, and has no side effects.

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

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

[0015] Furthermore, the probiotics are Lactobacillus plantarum powder, and the bacterial content in the powder can be 1*10 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 extract, and 5-12 parts of Gardenia jasminoides extract.

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

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

[0019] In another preferred embodiment of the present invention, the TCM compound comprises the Ziziphus jujuba seed extract and the Gardenia jasminoides extract, and the mass ratio of the Ziziphus jujuba seed extract to the Gardenia jasminoides extract is 8:3.

[0020] The second aspect of the present invention provides a method for preparing a probiotic-TCM compound composition having a sleep-aiding effect, comprising the following steps:

[0021] S1. The extract raw material is extracted twice with water at an extraction temperature of 90 to 100 ° C, and then the impurities are filtered out to obtain an extract; the extract raw material is jujube seed powder, Poria powder, lily powder or gardenia powder;

[0022] S2. The extract is concentrated under vacuum to a relative density of 1.09 to 1.12 g / cm 3 , then add maltodextrin and sterilize by high pressure;

[0023] S3. In a clean environment, the sterilized extract is spray-dried to obtain a powdered material; the powdered material is crushed, sieved, and mixed to obtain an extract powder;

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

[0025] Furthermore, in step S1, when the extract raw material is spiny jujube seed powder, 8 times and 6 times of water are used to soak the spiny jujube seed powder, and the extraction time is 120 minutes and 60 minutes respectively;

[0026] When the extract raw material is Poria powder, 10 times and 8 times of water are used to soak the Poria powder, and the extraction time is 120 minutes and 60 minutes respectively;

[0027] When the extract raw material is lily powder, 8 times and 8 times of water are used to soak the lily powder, and the extraction time is 120 minutes and 60 minutes respectively;

[0028] When the extract raw material is gardenia powder, 8 times and 6 times water are used to soak the gardenia powder, and the extraction time is 120 minutes and 60 minutes respectively.

[0029] Furthermore, in step S2, the vacuum degree is controlled to be -0.06 to -0.09 MPa during the concentration, and the concentration temperature is 50-75°C; the temperature is controlled to be 115-135°C during the high-pressure sterilization, and the time is 15-30 minutes.

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

[0031] Further, taking the Chinese date seed extract as an example, the specific extraction method is as follows:

[0032] a1 Source of raw materials and preliminary testing: After receiving the raw materials of sour jujube kernel powder, they are strictly tested by the quality control (QC) department to ensure that they meet the standards. The raw materials that pass the inspection enter the extraction process.

[0033] a2 Extraction and filtration: Water extraction twice: the Ziziphus jujuba seed powder is extracted with 8 times water and 6 times water respectively, the first extraction time is 120 minutes, and the second extraction time is 60 minutes. The temperature is kept at 90℃ to 100℃ during the extraction process, and stirring is continued to ensure the full extraction of the active ingredients; Filtration: After each extraction is completed, the extract is filtered through a filter cloth or filter to remove insoluble impurities, ensuring that the extract is clear and contains high-purity active ingredients.

[0034] a3 Vacuum concentration and maltodextrin addition: Concentration conditions: The filtered extract is processed by vacuum concentration equipment, the vacuum degree is controlled at -0.06 to -0.09MPa, the temperature is set at 50℃ to 75℃, and concentrated to a relative density of 1.09 to 1.12 to ensure that the extract is within the optimal concentration range; Add maltodextrin: Add an appropriate amount of maltodextrin (according to the specified ratio) to the concentrated extract to enhance the stability and finished product characteristics of the product. The concentrate continues to be heated and stirred evenly to completely dissolve the maltodextrin.

[0035] a4 High temperature sterilization and drying: High pressure sterilization: The extract with maltodextrin is sterilized at 115℃ to 135℃ for 15-30 minutes to ensure that the microorganisms in the extract are completely inactivated; Spray drying: The sterilized extract is spray dried in a clean production area (Grade D) (the inlet air temperature is controlled at 180℃ to 205℃, and the outlet air temperature is 85℃ to 105℃). After drying, a powdered product is obtained, and 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 kernel powder needs to go through a crushing process to ensure that it reaches a uniform particle size; Sieving: Use a 100-mesh screen to preliminarily sieve the powder to remove larger particles. Then sieve again under the CCP (critical control point) standard to ensure the consistency of the product particle size.

[0037] a6 Mixing and packaging: Mixing: The qualified jujube seed powder is evenly mixed in proportion for 50 to 60 minutes to ensure the uniformity and stability of the powder; Internal packaging inspection: The mixed powder is subjected to internal control inspection to confirm that all indicators are qualified. The jujube seed powder that passes the inspection is sealed in the inner packaging and then packaged in the outer packaging; Finished product inspection and warehousing: The packaged finished product is inspected by a metal foreign body detector (CCP3). After passing the inspection, it is labeled with the product name, batch number and net content, and is stored in the warehouse for use.

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

[0039] The third aspect of the present invention provides the use of the probiotic-TCM compound composition in the preparation of a sleep-aiding preparation.

[0040] Furthermore, the sleep-aiding preparation is used to increase the inhibitory neurotransmitter content in the brain, reduce the excitatory neurotransmitter content, and reduce the expression of GFAP protein.

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

[0042] Compared with the prior art, the present invention has the following beneficial effects:

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

[0044] 2. The probiotic-TCM compound composition provided by the present invention has no preservatives and no artificial synthetic chemical components. While having the effect of helping sleep, it is highly safe, has no toxic side effects, and is suitable for long-term use. BRIEF DESCRIPTION OF THE DRAWINGS

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

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

[0047] Figure 3 This is a diagram showing the effects of probiotics combined with TCM compound on neurotransmitters in mouse brain tissue;

[0048] Figure 4 This is 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 group 1; D: TCM compound group 2; E: probiotic + TCM compound group 1; F: probiotic + TCM compound group 2; G: positive control group. DETAILED DESCRIPTION

[0050] The present invention is further described below in conjunction with specific embodiments so that those skilled in the art can better understand the present invention and implement it, 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 those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

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

[0053] Example 1

[0054] This embodiment provides a probiotic-TCM compound composition with a sleep-aiding effect, which comprises 3 parts of TCM compound and 40 parts of probiotics by weight;

[0055] The TCM compound comprises a jujube seed extract, a tuckahoe extract, a lily extract and a gardenia extract in a mass ratio of 8:1:3:3, which is recorded as TCM compound group 1. The probiotic is Lactobacillus plantarum P-8 powder.

[0056] Example 2

[0057] This embodiment provides a probiotic-TCM compound composition with a sleep-aiding effect, which comprises 3 parts of TCM compound and 40 parts of probiotics by weight;

[0058] The TCM compound comprises a spinulosa jujuba seed extract and a gardenia jasmine extract in a mass ratio of 8:3, which is recorded as TCM compound group 2. The probiotics are Lactobacillus plantarum P-8 powder.

[0059] Comparative Example 1

[0060] This embodiment provides a TCM compound composition with sleep-aiding effect, including Ziziphus jujuba seed extract, Poria cocos extract, Lilium extract and Gardenia jasminoides extract, and their mass ratio is 8:1:3:3, which is recorded as TCM compound group 1.

[0061] Comparative Example 2

[0062] This embodiment provides a TCM compound composition with a sleep-aiding effect, comprising a Ziziphus jujuba seed extract and a Gardenia jasminoides extract, wherein the mass ratio of the two is 8:3, and is recorded 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: 84 ICR mice aged four weeks were housed in an SPF (pathogen-free) laboratory at a temperature of 22-24°C, 12-h light-dark cycle, relative humidity of 40%-55%, and good ventilation. The animals were allowed free access to food and water. The animals were fed for 1 week before the experiments were carried out, and the animals were kept for 30 days in the subsequent experiments.

[0068] 1.2. Animal grouping: 84 mice were randomly divided into 7 groups, with 12 mice in each group, namely 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 groups of mice were raised normally and administered drugs according to the following methods:

[0071] (1) Blank group: 0.1 mL of normal saline was intragastrically administered;

[0072] (2) The probiotic group was intragastrically administered with 11 mg / d / pcs of probiotic dry powder of comparative example 3 every day;

[0073] (3) TCM1 group added the TCM compound of comparative example 1 to the feed every day (TCM compound addition amount was 0.38 g / 100 g, equivalent to 11 mg / d / pcs per head);

[0074] (4) TCM group 2 added the TCM compound of comparative example 2 to the feed every day (TCM compound addition amount was 0.38 g / 100 g, equivalent to 11 mg / d / pcs per head);

[0075] (5) The probiotics + TCM1 group added the TCM compound of Example 1 to the feed every day, and gavaged the probiotic dry powder quantitatively (TCM compound added at 0.21 g / 100 g, equivalent to 6 mg / d / pcs per head, probiotic dry powder 5 mg / d / pcs);

[0076] (6) The probiotics + TCM2 group added the TCM compound of Example 2 to the feed every day, and the probiotic dry powder was quantitatively gavaged (TCM compound added at a dosage of 0.18 g / 100 g, equivalent to 5.2 mg / d / pcs per head, probiotic dry powder 5.8 mg / d / pcs);

[0077] (7) The positive control group was treated with diazepam by oral gavage every day at a dosage of 2 mg / kg / d.

[0078] 1.4. Behavioral analysis

[0079] Before the formal experiment, the pre-experimental treatment was performed to determine the dose of pentobarbital sodium (40-60 mg / kg) that made the animals fall asleep 100% but did not make them sleep for too long. This dose was used for the formal experiment. The sleep status of mice in each group was observed after administration. If the mice did not fall asleep directly 30 minutes after administration after the last oral gavage, a subsequent collaborative sleep behavior experiment was conducted with a subthreshold dose of pentobarbital sodium (six mice in each of the 7 groups of 12 mice were randomly selected to avoid false positives.

[0080] 1.5. Physiological and biochemical analysis

[0081] The weight changes of mice were evaluated. When there was no significant weight loss and the mice were ethically approved, the mice were killed (the remaining 6 mice in the 7 groups) and the organ-to-body ratio was analyzed.

[0082] 1.6. Neurotransmitter analysis and immunofluorescence staining

[0083] Neurotransmitters were separated and extracted from brain tissue homogenates of sacrificed mice by 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 permeabilized with 0.1% Triton X-100. Samples were incubated with anti-GFAP antibody, GFAP was labeled with Alexa Fluor 488-conjugated secondary antibody, and nuclear staining was performed with Hoechst 33342. Analysis was visualized under a fluorescence microscope.

[0085] 2. Experimental results

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

[0087] The weight of mice reflects whether the mice grow healthily. Table 1 shows the effect of plant lactobacillus combined with TCM compound on the weight of mice. As can be seen from the table, after 30 days of intragastric administration, there was no significant difference in the weight of mice in each group (P>0.05). This shows that the probiotics combined with TCM compound has no side effects on the weight of mice and is safe.

[0088] Table 1

[0089] Group Initial body weight (g) Last weight (g) Blank Group <![CDATA[34.22±1.97 a ]]> <![CDATA[36.98±2.75 a ]]> Probiotics 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 of mice

[0091] As shown in Table 2, 30 days after intragastric administration, there were no significant differences in organ coefficients of the mice in each group, such as lung, kidney, spleen, liver, etc. (P>0.05), indicating that the probiotics combined with TCM compound had no side 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 into the category of improving sleep. When no animals fall asleep in the blank control group and the probiotics combined with TCM compound group, it can be determined that the probiotics combined with TCM compound has no inhibitory effect on the central nervous system, and the remaining three behavioral experiments are performed.

[0096] As shown in Table 3, 30 minutes after the last intragastric administration, there was no direct sleep phenomenon in both the blank control group and the probiotics combined with TCM compound group, and there was no significant difference (P>0.05), indicating that the probiotics combined with TCM compound could not make mice sleep directly.

[0097] Table 3

[0098] Group Experimental animals (only) Sleeping animals (only) Blank Group 12 <![CDATA[0 a ]]> Probiotics 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 duration in mice

[0100] The length of sleep time is directly related to sleep quality. Sodium pentobarbital is a drug that acts on the central nervous system, has a hypnotic effect, and can effectively prolong sleep time. Compared with the blank control group, if the sleep time of mice in the probiotics combined with TCM compound group is significantly prolonged, it means that the sample can synergize with sodium pentobarbital to prolong the sleep time of mice.

[0101] like Figure 1 As shown in the figure, compared with the blank control group, the sleep time of the probiotics combined with TCM compound 2 group was prolonged by 34.37 minutes. Therefore, the probiotics combined with TCM compound 2 group can enhance the effect of sodium pentobarbital, improve sleep, and prolong the sleep time of mice.

[0102] 2.5. Effects of Lactobacillus plantarum combined with TCM compound on the hypnotic experiment of pentobarbital sodium in mice

[0103] Sodium pentobarbital is a drug that acts on the central nervous system and can be metabolized by liver enzymes. If the sample has an inhibitory effect on liver enzymes, the sleep time will also be prolonged, which will result in a false positive result. In order to eliminate the possibility of false positives, a subthreshold dose hypnosis experiment of sodium pentobarbital should be conducted again. After the last administration, each mouse was intraperitoneally injected with sodium pentobarbital; the injection volume was 0.2mL / 20g (mouse), and the disappearance of the righting reflex was used as an indicator.

[0104] As shown in Table 4, compared with the blank control group, the sleep incidence of mice in each group was increased. Among them, the sleep incidence of probiotics combined with TCM 1 group and probiotics combined with TCM 2 group were 66.67% and 91.67%, respectively, with significant differences (P<0.05). The results show that it can synergize with sodium pentobarbital to increase the sleep incidence.

[0105] Table 4

[0106] Group Experimental animals (only) Sleeping animals (only) Sleep incidence (%) Blank Group 12 2 <![CDATA[16.67% e ]]> Probiotics 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 barbiturate sodium sleep latency in mice

[0108] The speed of falling asleep is also an important indicator of sleep. A short sleep latency can indicate a state of rapid sleep. Sodium barbital is also a drug that has an inhibitory effect on the central nervous system. After intraperitoneal injection, if the sleep latency in the probiotics combined with TCM compound group is significantly shortened, that is, the sample and sodium barbital also have a synergistic effect, the experimental result is positive.

[0109] Depend on Figure 2 As shown in the figure, compared with the blank control group, the sleep time of probiotics combined with TCM compound group 1 and probiotics combined with TCM compound group 2 was shortened. Among them, probiotics combined with TCM compound group 2 had the best effect on improving sleep, and the sleep time was shortened by 13 minutes. The results showed that probiotics combined with TCM compound had a synergistic effect with sodium barbital and could shorten the sleep latency of mice.

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

[0111] See also Figure 3 Compared with the blank group and the positive control group, the inhibitory neurotransmitter content in probiotics combined with TCM compound group 1 and group 2 increased. Compared with the blank group, the excitatory neurotransmitter content in probiotics combined with TCM compound group 1 and group 2 was significantly reduced (P < 0.05). This shows that the probiotic-TCM compound composition can effectively increase the 5-HT and GABA content in the mouse brain and reduce the DA content, 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, the number of which is 10-50 times that of neurons. Among them, astrocytes (AS) are cells that are closely related to sleep. Studies have shown that astrocytes can maintain the sleep-wake cycle by releasing ATP in the brain. Glial fibrillary acidic protein (GFAP) is a type III intermediate filament protein that exists in monomeric form and is a marker of AS activation.

[0114] Depend on Figure 4 It can be seen that the green area represents GFAP. When in a sleep disorder state, AS begins to activate and the expression of GFAP increases. Compared with the blank group, the expression of GFAP in probiotics combined with TCM compound group 1 and group 2 was downregulated, indicating that probiotics combined with TCM compound can assist sleep by regulating GFAP expression.

[0115] In summary, the probiotic-TCM compound composition has no effect on the weight and organ coefficient of mice, is non-toxic, and is safe. Based on the results of epibehavioral studies, the probiotic-TCM compound composition has no direct hypnotic effect, can prolong sleep time, shorten sleep latency, and increase sleep incidence. The experimental results are all positive, proving that the probiotic-TCM compound composition has the effect of improving sleep. At the same time, based on brain tissue biochemistry and histopathological analysis, it was found that the probiotic-TCM compound composition improves sleep by increasing the content of inhibitory neurotransmitters in the mouse brain, reducing the content of excitatory neurotransmitters, and reducing the expression of GFAP protein. Therefore, the probiotic-TCM compound composition is effective, safe, and has no side effects in assisting sleep.

[0116] The above-described embodiments are only preferred embodiments for fully illustrating the present invention, and the protection scope of the present invention is not limited thereto. Equivalent substitutions or changes made by those skilled in the art based on the present invention are within the protection scope of the present invention. The protection scope of the present invention shall be subject to the claims.

Claims

1. A probiotic-TCM compound composition with sleep-aiding effect, characterized in that: Includes TCM compound and probiotics; The TCM compound comprises a Chinese date seed extract, a tuckahoe extract, a lily extract and a gardenia extract; The probiotic is Lactobacillus plantarum.

2. A probiotic-TCM compound composition with sleep-aiding effect according to claim 1, characterized in that: 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 extract, and 5-12 parts of Gardenia jasminoides extract.

3. A probiotic-TCM compound composition with sleep-aiding effect according to claim 2, characterized in that: In the TCM compound, the mass ratio of the Ziziphus jujuba seed extract, the Poria cocos extract, the lily extract and the gardenia extract is 8:1:3:

3.

4. The probiotic-TCM compound composition with sleep-aiding effect according to claim 2, characterized in that: The TCM compound comprises a Chinese jujube seed extract and a gardenia extract, and the mass ratio of the Chinese jujube seed extract to the gardenia extract is 8:

3.

5. The probiotic-TCM compound composition with sleep-aiding effect according to claim 1, characterized in that: By weight, the composition comprises: 3 to 8 parts of probiotics and 2 to 5 parts of TCM compound; or, The probiotics are Lactobacillus plantarum powder, and the bacterial content is 1*10 10 CFU / g.

6. The method for preparing a probiotic-TCM compound composition having a sleep-aiding effect according to any one of claims 1 to 5, characterized in that: The following steps are involved: S1. The extract raw material is extracted twice with water at an extraction temperature of 90 to 100 ° C, and then the impurities are filtered out to obtain an extract; the extract raw material is jujube seed powder, Poria powder, lily powder or gardenia powder; S2. The extract is concentrated under vacuum to a relative density of 1.09 to 1.12 g / cm 3 , then add maltodextrin and sterilize by high pressure; S3. In a clean environment, the sterilized extract is spray-dried to obtain a powdered material; the powdered material is crushed, sieved, and mixed to obtain an extract powder; S4. Mixing the extract powder with probiotic powder to obtain the probiotic-TCM compound composition.

7. The method for preparing a probiotic-TCM compound composition having a sleep-aiding effect according to claim 6, characterized in that: In step S1, when the extract raw material is spiny jujube seed powder, 8 times and 6 times of water are used to soak the spiny jujube seed powder, and the extraction time is 120 minutes and 60 minutes respectively; When the extract raw material is Poria powder, 10 times and 8 times of water are used to soak the Poria powder, and the extraction time is 120 minutes and 60 minutes respectively; When the extract raw material is lily powder, 8 times and 8 times of water are used to soak the lily powder, and the extraction time is 120 minutes and 60 minutes respectively; When the extract raw material is gardenia powder, 8 times and 6 times water are used to soak the gardenia powder, and the extraction time is 120 minutes and 60 minutes respectively.

8. The method for preparing a probiotic-TCM compound composition having a sleep-aiding effect according to claim 6, characterized in that: In step S2, the vacuum degree is controlled to be -0.06 to -0.09 MPa during concentration, and the concentration temperature is 50-75° C.; the temperature is controlled to be 115-135° C. during high pressure sterilization, and the time is 15-30 minutes; In step S3, during spray drying, the air inlet temperature is 180-205°C, and the air outlet temperature is 85-105°C.

9. Use of the probiotic-TCM compound composition according to any one of claims 1 to 5 in the preparation of a sleep-aiding preparation.

10. The use according to claim 9, characterized in that: The sleep-aiding preparation is used to increase the inhibitory neurotransmitter content in the brain, reduce the excitatory neurotransmitter content, and reduce the expression of GFAP protein; and / or, The preparations include foods, medicines and health products.

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