WO2025129426A1 - Microorganism/grain/herb fermentation compound capable of delaying aging and aiding in sleep by cerebral dopamine production, method for preparing same, and use thereof - Google Patents

Abstract

Provided are a microorganism/grain/herb fermentation compound with the effects of delaying aging and aiding in sleep, a method for preparing same, and use thereof. The compound is prepared from Gastrodia elata rhizoma, black rice, and wheatgrass by means of fermentation, and can promote dopamine production, help maintain neural stability in brains, delay brain aging, protect brain nerves, and calm the nerves to help sleep.

Description

A fermented compound of Spartina officinalis that can delay aging and promote sleep through brain dopamine production, and its preparation method and application Technical Field

The present invention provides a smilax glabra fermentation compound that can delay aging and soothe sleep through brain dopamine production, which can effectively promote the human body to produce dopamine, glutamic acid and vitamin B6, thereby delaying brain aging, protecting brain nerves and calming nerves to help sleep.

Background Art

Dopamine is an important neurotransmitter that can affect a person's mood. It is an organic compound that plays several important roles in the brain and body. It is usually synthesized in the human brain and kidneys. In the brain, dopamine acts as a neurotransmitter, sending signals to other nerve cells through the release of chemicals by neurons. Several important neurological diseases are associated with dysfunction of the dopamine system, so some key drugs that can modify the role of dopamine can be used to treat them. For example, Parkinson's disease: a degenerative condition that causes body tremors and movement disorders is caused by insufficient secretion of dopamine by secretory neurons in a region of the midbrain called the substantia nigra. Its metabolic precursor L-dopa (L-DOPA) can be manufactured industrially, and levodopa is the most widely used treatment. There is evidence that schizophrenia involves changes in dopamine activity levels, and most commonly used antipsychotic drugs have the primary effect of reducing dopamine activity. In addition, restless legs syndrome and attention deficit hyperactivity disorder are associated with reduced dopamine activity. Dopamine itself can be made into an intravenous drug. Although it cannot reach the brain from the blood, its peripheral effects make it useful for the treatment of heart failure or shock, especially for newborn babies. In addition, studies have found that moderate dopamine can help the quality of sleep.

Glutamic acid (GA) is an amino acid found in natural proteins. Since glutamate can be synthesized in the human body, it is classified as a non-essential amino acid. Glutamate is commonly found in animals and plants in various forms. It is also the most abundant excitatory neurotransmitter in the nervous system of vertebrates and a precursor of γ-aminobutyric acid (GABA).

Vitamin B6 (Vitamin B), also known as anti-dermatitis vitamin and pyridoxine, is a type of B vitamin and an essential vitamin. It is composed of six vitamin isomers that can be converted into each other. It is closely related to amino acid metabolism and is an amino acid. A coenzyme for amino acid decarboxylase, transaminase, etc. The most common chemical form is pyridoxine; the most biologically active is pyridoxal phosphate, which acts as a coenzyme in more than 140 enzyme reactions in the metabolism of amino acids, glucose and lipids. Plants synthesize pyridoxine to protect against ultraviolet B in sunlight and synthesize chlorophyll, but animals cannot synthesize vitamin B6, so they need to obtain vitamin B6 by eating plants or other animals; although intestinal bacteria also produce some vitamin B6, it is not enough to meet the needs of animals. For adults, the recommended dietary intake of vitamin B6 is 1.0 to 2.0 mg per day, and the safe upper limit is 25 to 100 mg per day. Vitamin B6 deficiency is rare, and common symptoms include rash and inflammation of the mouth and eyes, drowsiness, and peripheral neuropathy that affects the sensory and motor nerves of the hands and feet; in addition, there are dermatitis, spasms, anemia, etc. At the same time, some rare genetic diseases can cause infants to have epileptic seizures due to vitamin B6 deficiency.

Gastrodia elata is a perennial saprophytic upright herb with underground rhizomes in the shape of tubers. It is a precious medicinal material and is recorded in many Chinese pharmacopoeias. For example, in Compendium of Materia Medica, it is recorded that "Gastrodia elata is a medicine for the liver meridian and qi". Medicinal Gastrodia elata is the dried tuber of the plant Gastrodia elata. It is long oval or long strip, wrinkled, has a unique smell, tastes sweet and slightly pungent, and is mainly distributed in Sichuan, Yunnan and other places. Gastrodia elata is sweet, pungent, flat, non-toxic in nature and belongs to the liver meridian. It can calm wind and stop spasms, calm liver yang, dispel wind and dredge collaterals. It has the effect of treating convulsions, convulsions, cramps, dizziness, headaches, hemiplegia, limb numbness, rheumatic pain, etc. It can be taken orally in a decoction of 3-10 grams, or made into pills, powders, or ground into powder and swallowed, 1-1.5 grams each time, but it should be used with caution in patients with severe qi and blood deficiency.

Purple rice, also known as black rice, is a type of purple-black rice. Purple rice can hybridize with other rice species, so there is no reproductive isolation from other rice species, and they all belong to rice. In addition, black rice was also called "forbidden rice" in ancient China, because generally speaking, only people in the upper class could afford it. The activity of hydrophilic antioxidants in purple rice bran is much higher than that of lipophilic antioxidants, anthocyanins and γ-stanols, which are mainly located inside the purple rice bran. Its lipophilic antioxidant extract containing gamma-oryzanol has anti-inflammatory activity.

Wheatgrass/Catgrass is a member of the genus Agropyron, especially the young shoots of Agropyron cristatum (a cousin of wheat). Its young leaves can be squeezed into juice or dried and ground into powder. Unprocessed plants contain a large amount of cellulose that is difficult for humans to digest, but they also contain beneficial ingredients such as chlorophyll, amino acids, vitamins, minerals and enzymes. In addition, the Compendium of Materia Medica records the therapeutic effects of wheatgrass, "Wheatgrass seedlings have a pungent, cold smell and are non-toxic. They are mainly used to eliminate alcohol poisoning, sudden fever, alcohol carbuncle, yellow eyes... Mash it and squeeze out the juice to drink it every day, it will quench thirst, relieve chest heat and insulate, and benefit the small intestine..." This shows that wheatgrass has the effects of reducing inflammation and fever, strengthening the stomach and digestion, and dredging the stomach.

In the past, Gastrodia elata was mainly used as an additive in the form of Gastrodia elata extract. However, no research has confirmed that Gastrodia elata alone can actually stimulate the body's own production of dopamine or help the brain maintain nerve stability. Therefore, additional ingredients such as phosphatidylserine and vitamins are also added to increase the effect of stabilizing brain nerves.

In view of this, the relevant field urgently needs to develop a Gastrodia elata fermentation complex that can truly promote dopamine and help the brain maintain neural stability, and to confirm through experiments that it can delay brain aging, protect brain nerves, calm nerves and help sleep.

Summary of the invention

In order to solve the problem that the above-mentioned natural plant extracts cannot effectively help the brain maintain nerve stability, the present invention utilizes natural food ingredients that are mixed in a specific proportion and fermented with probiotics to increase the dopamine content and the concentration of antioxidants SOD, GPx, and G6PD in brain tissue, and reduce the content of peroxides 8-oxodG and MDA in brain tissue, which has the potential to delay brain aging and protect brain nerves.

In one embodiment of the present invention, a method for preparing a Japonica foetida fermentation complex that can delay aging and promote sleep through brain dopamine production comprises: a Japonica foetida raw material fragmentation stage, a negative pressure wall breaking stage, an extraction fermentation stage, and a modification fermentation stage.

The subsidiary technical means derived from the above necessary technical means are, among others, the crushing stage of the raw material of the spatholobi is to obtain multiple spatholobi crude extracts by respectively pressing multiple (plural) spatholobi raw materials.

Preferably, the raw materials of the rice grass are gastrodia elata fruiting bodies, black rice and wheat grass.

Preferably, the plurality of crude extracts of Gastrodia elata fruiting bodies are crude extracts, Gastrodia elata fruiting body residues, crude black rice extracts, black rice residues, crude wheat grass extracts, and wheat grass residues.

The auxiliary technical means derived from the above necessary technical means are: in the negative pressure wall breaking stage, the crude extract of multiple fungi and spatholobi is placed in effect The first extract was obtained on the day.

The subsidiary technical means derived from the above necessary technical means are that, in the refined fermentation stage, 0.1-0.5% (w/w) pectinase and the first extract are implanted with 0.2-2% (w/w) lactic acid bacteria; and in the refined fermentation stage, the fermentation temperature is controlled at The fermentation is continued for 8 to 14 days to obtain the first fermentation liquid.

The subsidiary technical means derived from the above necessary technical means are: in the modified fermentation stage: 0.2-2% (w/w) yeast or acetic acid bacteria is implanted into the first fermentation liquid; in the modified fermentation stage, the fermentation temperature is controlled at Continuous fermentation The second fermentation liquid obtained on the day.

In one embodiment of the present invention, the Spartina umbellata fermentation complex comprises a plurality of Spartina umbellata crude extracts, lactic acid bacteria, and yeast or acetic acid bacteria.

Preferably, the plurality of crude extracts of Gastrodia elata fruiting bodies are crude extracts of Gastrodia elata fruiting bodies, residues of Gastrodia elata fruiting bodies, crude extracts of black rice, residues of black rice, crude extracts of wheat grass, and residues of wheat grass.

In one embodiment of the present invention, the Spartina officinalis fermentation complex has the function of increasing the number and activity of lactic acid bacteria that synthesize gamma-aminobutyric acid (GABA) in the intestines of mammals.

In one embodiment of the present invention, the Spartina officinalis fermentation complex improves the sleep quality of an individual and can maintain the average deep sleep time of the individual within a normal range.

Preferably, the fermented compound of Junmi grass can restore the proportion of oculomotor period to

In one embodiment of the present invention, the Spartina officinalis fermentation complex has the efficacy of preventing and/or treating Parkinson's disease.

In one embodiment of the present invention, the Spartina officinalis fermentation complex has the effect of enhancing the anti-oxidation of the brain.

In one embodiment of the present invention, the Spartina officinalis fermentation complex has a nerve-protecting effect.

In one embodiment of the present invention, the Spartina officinalis fermentation complex has the efficacy of treating nerve damage.

In one embodiment of the present invention, the Spartina officinalis fermentation complex has the effect of preventing the decrease of dopamine in the brain.

In one embodiment of the present invention, the Spartina officinalis fermentation complex has the efficacy of treating decreased dopamine in the brain.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 shows the changes in sugar content of different strains during the refined fermentation stage of the present invention;

FIG2 shows the pH value changes of different strains during the refined fermentation stage of the present invention;

FIG3 shows the changes in sugar content of different strains during the modified fermentation stage of the present invention;

FIG4 shows the pH value changes of different strains during the modified fermentation stage of the present invention;

FIG5 shows the sleep-inducing effect of the Spartina officinalis fermentation complex of the present invention, which can delay aging and soothe sleep through the production of brain dopamine;

FIG6 shows the effect of the Spartina officinalis fermentation complex of the present invention on promoting the content of dopamine in the brain through the production of brain dopamine to delay aging and promote sleep;

FIG. 7 shows the effect of the Spartina officinalis fermentation compound of the present invention on tyrosine hydroxylase in brain tissue slices, which can delay aging and promote sleep through brain dopamine production;

FIG8 shows the effect of the Spartina officinalis fermentation complex of the present invention, which can delay aging and promote sleep through brain dopamine production, on the activity of the antioxidant enzyme G6PD in brain tissue;

FIG. 9 shows the effect of the Spartina officinalis fermentation complex of the present invention, which can delay aging and promote sleep through brain dopamine production, on the activity of antioxidant GPx in brain tissue;

FIG. 10 shows the effect of the Spartina officinalis fermentation complex of the present invention, which can delay aging and promote sleep through brain dopamine production, on the activity of superoxide dismutase (SOD) in brain tissue;

FIG. 11 shows the effect of the Spartina officinalis fermentation complex of the present invention, which can delay aging and promote sleep through brain dopamine production, on the content of the oxide 8-oxodG in brain tissue;

FIG. 12 shows the effect of the Spartina officinalis fermentation complex of the present invention, which can delay aging and promote sleep through brain dopamine production, on the content of lipid peroxide MDA in brain tissue.

DETAILED DESCRIPTION

The technical content of the present invention is described below through specific embodiments, and people familiar with this art can understand the advantages and effects of the present invention from the contents disclosed in this specification. However, the present invention can also be implemented or applied through other different specific embodiments.

Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by those skilled in the art to which the present invention belongs. Those skilled in the art will recognize that many methods and materials similar or equivalent to those described herein can be used to implement the present invention. Of course, the present invention is in no way limited to the methods and materials described.

The above embodiments are only for explaining the principle and efficacy of the present invention, and their purpose is to enable those familiar with the above technology to understand the content of the present invention and implement it accordingly, but they are not intended to limit the present invention. Therefore, those familiar with this technology can make equivalent modifications, changes and variations to the above embodiments without departing from the spirit of the present invention. The scope of rights of the present invention shall be as listed in the claims of the above patent application.

Example 1: Preparation of the Spartina officinalis fermentation complex

Gastrodia elata, black rice and wheat sprouts are combined with lactic acid bacteria, yeast or acetic acid bacteria to carry out stage fermentation steps, and the fermentation liquid is filtered and retained after the fermentation is completed; the stage fermentation steps include:

(1) Crushing of the raw materials of Junmi grass: the wheat seedlings are squeezed physically until they are crushed, while retaining the juice, pomace or leaf residue; the gastrodia elata and black rice are soaked in hot water at 70-100° C. for 30 minutes to soften, and then squeezed until the fruiting bodies are in pieces, while retaining the juice and residue;

(2) Negative pressure cell wall breaking stage: organic brown sugar, isomaltooligosaccharide, sorbitol, granulated sugar, sucrose or a combination thereof is added at 0.2-1% (w/w) to increase the osmotic pressure of the extract, and negative pressure extraction is performed in a near-vacuum environment of 20 cmHg to 60 cmHg for 5-14 days to break the cell walls of fruits and vegetables and release intracellular nutrients and intracellular polysaccharides;

(3) Extract fermentation stage: lactic acid bacteria (Lactobacillus plantarum, Lactobacillus delbrueckii, Lactococcus lactis, Lactobacillus acidophilus or Bifidobacterium (B.bifidum)) or a combination thereof is implanted into the extract at a ratio of 0.2-2% (w/w), the fermentation temperature is controlled at 22-28°C, and the fermentation is continued for 8-14 days. During this stage, the decomposition characteristics of lactic acid bacteria are utilized to produce a variety of decomposition enzymes to degrade the nutrients of fruits and vegetables into small molecules. During the fermentation process, 0.1-0.5% (w/w) pectin decomposition enzyme is added to increase the decomposition rate;

(4) Modification fermentation stage: one or a combination of yeast or acetic acid bacteria (S. fibuligera, S. cerevisiae, Pichia faciens, S. pombe, A. hansenii, A. xylinum, A. suboxydans) is implanted into the extract at a ratio of 0.2-2% (w/w), the fermentation temperature is controlled at 22-28°C, and the fermentation is continued for 10-21 days. In this stage, the decomposition characteristics of microorganisms are utilized to produce a variety of decomposition enzymes to decompose the polysaccharide components into small molecules, the viscosity of the fermentation liquid is reduced and the fluidity is improved. After the fermentation is completed, the mycelium fermentation complex is obtained for use in the subsequent experiments of the implementation examples.

Example 2: Comparison of stage fermentation tests

In order to test the influence of strains on the fermentation effect at different fermentation stages, so as to obtain the best fermentation result, the present invention further uses different strains to carry out fermentation experiments. In the embodiment, two different strains (Lactobacillus plantarum (LP) and Lactobacillus bulgaricus (LD) are used in the extract fermentation stage; Aspergillus fibuligera (SF) and Saccharomyces cerevisiae (SC) are used in the modified fermentation stage) to carry out fermentation comparison and evaluate the best fermentation strain. During the fermentation process, the change range of sugar content and pH value is proportional to the fermentation activity of the strain. In short-term fermentation, the activity of the strain can greatly accelerate the completeness of the fermentation. The results are shown in Table 1, Table 2 and Figures 1-4. It can be seen that different strains do have a great impact on the fermentation effect. In the essence fermentation stage, Lactobacillus plantarum can more effectively enhance the decomposition effect than Lactobacillus delbrueckii, while in the modified fermentation stage, Aspergillus fibuligera can better reduce the viscosity of the fermentation liquid and improve the fluidity than Saccharomyces cerevisiae.

Example 3: Establishment of Parkinson's disease animal model

Establishment of Parkinson's model: The mice were divided into 4 groups: male mice, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induced control group (Ctrl), MPTP induced and given a low dose of Spartina umbellata fermentation complex (L), MPTP induced and given the recommended dose of Spartina umbellata fermentation complex (M) and MPTP induced and given a high dose of Spartina umbellata fermentation complex (H); female mice, 1 group, MPTP induced and given a high dose of Spartina umbellata fermentation complex (F), with 12 mice in each group. The groups that were given the Ferment Complex of Spartina umbellata were all given the Ferment Complex of Spartina umbellata by tube feeding for 28 consecutive days. The low-dose Ferment Complex of Spartina umbellata was 0.195 mg per gram of mouse body weight, equivalent to 250 mg/70 kg/day for humans; the recommended dose of Ferment Complex of Spartina umbellata was 0.39 mg per gram of mouse body weight, equivalent to 500 mg/70 kg/day for humans; the high-dose Ferment Complex of Spartina umbellata was 1.17 mg per gram of mouse body weight, equivalent to 750 mg/70 kg/day for humans.

Example 4: Experiment on the self-production of GABA in the intestine

The intestinal environment (pH 8.3) was simulated in NaOH solution, and intestinal probiotics were added to simulate intestinal bacteria and environment. 750 mg of the Spartina fermentation complex was added to 50 ml of NaOH aqueous solution, and the solution was taken out every hour to detect the number of bacteria and GABA content. In order to compare the difference between the Spartina fermentation complex of the present invention and the general Gastrodia extract, the Spartina extract complex formed by mixing Gastrodia, black rice and wheat seedling extracts in the same proportion of the Spartina fermentation complex was also tested. The results are shown in Tables 3 and 4 below.

Table 3. Experimental results of the intestinal simulation of GABA self-production by the Spartina spp. fermentation complex

Table 4. Results of the experiment on the simulated GABA self-generation in the intestine of the general Spartina officinalis extract complex

Comparison table three, table four can be seen, the present invention's bacterium rice grass fermentation complex under the simulated intestinal flora environment, intestinal release GABA precursor (fermentation type), can be through intestinal probiotics effect, promote self-generation GABA; when slowly released in the intestinal tract for more than 3 hours, the fourth hour GABA is synthesized in large quantities, for the human body to enter the best time of deep sleep, with staged pressure relief, sleep effect. GABA precursor will be slowly generated after entering the blood-brain barrier, and the generation time lasts for more than 8 hours to ensure that GABA is completely absorbed and utilized by the brain. However, the general bacterium rice grass extraction complex is that the GABA content is absorbed by the human body and the content reaches the highest peak within 0.5 hours of taking, and even surpasses the bacterium rice grass fermentation complex of the present invention, but with the increase of time, the content of GABA gradually decreases, representing that the bacterium rice grass extraction complex cannot effectively promote the self-generation of GABA in the human intestinal tract, but because the absorption effect causes the GABA content to increase with time and gradually decrease until it is exhausted, so the bacterium rice grass fermentation complex of the present invention and the general bacterium rice grass extraction complex that is not fermented have great differences in nature.

Embodiment 5: Human sleep monitoring

18 men or women aged 20 to 60 were given 750 mg of the fermented complex of Spartina officinalis 30 minutes before bedtime and were asked to wear smart wearable devices for 5 days of sleep monitoring. The sleep duration and the length of the REM sleep were monitored. The study pointed out that every 5% decrease in the REM sleep will increase the mortality rate by 17% and increase the risk of dementia.

The results are shown in Figure 5. Among the test groups, the deep sleep time of all test subjects was increased on the first day of use, and the average deep sleep time increased by 4.6% on the first day of use, and the REM sleep period was shortened by 5.09%. On the second day of use, the deep sleep time was slightly reduced, and the REM sleep period increased significantly, which means that the body function has gradually recovered on the second day, and the brain function has begun to strengthen and solidify memory. After the third day, the average deep sleep time can be maintained within the normal range, and the REM sleep period accounts for 20-25%, which is within the healthy range.

Example 6. Determination of dopamine concentration in the brain

Dopamine is mainly found in the human brain area, commonly known as the happy hormone, which controls emotions and stress changes. Moderate dopamine secretion helps sleep.

The experiment used drugs to induce young Parkinson's model mice. Male mice were tube-fed at three different doses and female mice were tube-fed at the highest dose. The mice were continuously given the fermentation complex of Spartina officinalis for 28 days before being sacrificed. After sacrifice, the striatum of the mice was removed to measure the dopamine concentration in the mouse brain in order to analyze the dopamine concentration in their brain tissue.

The results are shown in Figure 6. There is a significant difference in the substantia nigra injury mice after treatment with low doses and recommended doses, indicating that low doses and recommended doses have significant therapeutic effects, but there are still significant differences from healthy mice, so the evaluation can be used as a dose for preventive effect. After high-dose treatment of male and female substantia nigra injury mice, there is no significant difference from healthy mice, indicating that high doses have the effect of treating insufficient dopamine secretion. Therefore, in summary, low doses and recommended doses have the effect of preventing dopamine deficiency to achieve preventive effects, while high doses can be used as effective doses for treatment.

Example 7: Testing of the content of tyrosine hydroxylase, a precursor of dopamine in the brain

Tyrosine hydroxylase (TH) is considered the basis of dopamine secretion in brain tissue. When TH concentration decreases, dopamine production decreases, which can lead to sleep disorders, depression, and inattention.

The experiment used drugs to induce Parkinson's disease in young mice. Male mice were tube-fed with three different doses and female mice were tube-fed with the highest dose. The mice were continuously given the fermentation complex of Spartina officinalis for 28 days before being sacrificed. The brain tissues of the mice were removed for dehydration, embedding, sectioning and staining. TH immunostaining was performed and the pathological changes of dopamine precursor tyrosine hydroxylase (TH) in the brain tissues were observed under an optical microscope. The amount of TH was used as the basis for diagnosis and record evaluation.

The results are shown in Figure 7. There are differences in mice with substantia nigra injury after treatment with low doses, indicating that low doses can be used as preventive consumption in normal health care effects. There are significant differences in mice with substantia nigra injury after treatment with the recommended dose, and the therapeutic effect is significantly higher than that of the low dose group, but there are still significant differences compared with healthy mice. There is no significant difference between male and female mice with substantia nigra injury after treatment with high doses and healthy mice, indicating that high doses have the effect of preventing dopamine deficiency. Therefore, in summary, low doses and recommended doses have the effect of preventing substantia nigra injury, and high doses can be used as effective doses for treatment.

Example 8. Antioxidant Index Content in Blood

The aging of dopamine neurons is generally considered to be due to excessive oxidation, which causes neuronal oxidative atrophy. Therefore, the higher the antioxidant index, the stronger the ability to protect nerve cells. If the G6PD enzyme exists in brain tissue, it can produce antioxidant substances to resist the damage of oxidative free radicals. Studies have shown that if the G6PD enzyme activity is low in patients, the antioxidant capacity will be lost. SOD is a comprehensive free radical antioxidant enzyme. The activity of SOD can determine the antioxidant capacity of an organism, so it is commonly used as one of the antioxidant indicators.

The experiment used drug-induced aging Parkinson's model mice, feeding them three different doses for 12 weeks, and then taking blood samples from the mice to analyze the activity of the antioxidant indicators SOD, G6PD and GPx.

The results are shown in Figures 8-10. Whether it is the content of antioxidant indicators such as G6PD, GPx or SOD, there are significant differences in the substantia nigra injured mice after low-dose treatment, indicating that the low dose has a significant therapeutic effect, but there are still significant differences from healthy mice. There are significant differences in the substantia nigra injured mice after treatment with the recommended dose, and the therapeutic effect is significantly higher than the low-dose group, but there are still significant differences from healthy mice. After high-dose treatment of male and female substantia nigra injured mice, there is no significant difference from healthy mice, indicating that the high dose can prevent the problem of dopamine secretion deficiency. Therefore, in summary, low doses and recommended doses have the effect of preventing dopamine deficiency and protecting neurons, and high doses can be used as effective doses for treatment.

Example 9: Determination of brain tissue oxide concentration

Studies have shown that 8-oxodG is a strong free radical oxidant, which can cause neuron damage in the brain.

The experiment used drug-induced aging Parkinson's model mice. Male mice were fed three different doses and female mice were fed the highest dose. The mice were continuously given the fermentation complex of Spartina officinalis for 28 days, then sacrificed. The mouse brain tissue was removed and the mitochondria in the brain tissue were separated. The mitochondrial DNA was extracted and the 8-oxodg content in the brain mitochondrial DNA was analyzed to evaluate the degree of mitochondrial DNA damage.

The results are shown in Figure 11. After the mice with substantia nigra injury were treated with a low dose, there was a significant difference, indicating that the low dose had a significant therapeutic effect, but there was still a significant difference from healthy mice. After the mice with substantia nigra injury were treated with the recommended dose, there was a significant difference, and the therapeutic effect was significantly higher than the low-dose group, but there was still a significant difference from healthy mice. After high-dose treatment of male and female mice with substantia nigra injury, there was no significant difference from healthy mice, indicating that the high dose has a protective effect on neurons. Therefore, in summary, low doses and recommended doses have the effect of preventing neuronal damage, and high doses can be used as effective doses for treating neuronal damage.

Example 10: Determination of lipid oxide concentration in brain tissue

Malondialdehyde (MDA) is a lipid peroxide that can cause neuronal damage when present in the brain.

The experiment used drug-induced aging Parkinson's disease model mice, and fed male mice with three different doses and female mice with the highest dose. After 28 days of continuous administration of the Spartina fermentation complex, the mice were sacrificed and brain tissue samples were taken to analyze the content of MDA. Reactive oxygen species (ROS) can cause lipid peroxidation. The formation of MDA affects the oxidation reactions of cell membranes, lipoproteins, and other lipid-containing molecules, causing oxidative atrophy of neurons.

The results are shown in Figure 12. After the mice with substantia nigra injury were treated with a low dose, there was a significant difference, indicating that the low dose had a significant therapeutic effect, but there was still a significant difference compared with healthy mice. After the mice with substantia nigra injury were treated with the recommended dose, there was a significant difference, and the therapeutic effect was significantly higher than the low-dose group, but there was still a significant difference from healthy mice. After high-dose treatment of male and female mice with substantia nigra injury, there was no significant difference from healthy mice, indicating that the high dose has a neuroprotective effect. Therefore, in summary, low doses and recommended doses have the effect of protecting neurons, and high doses can be used as effective doses for treating damaged neurons.

The above experimental data are preliminary experimental results obtained under specific conditions, which are only used to facilitate understanding or reference of the technical content of the present invention, and other related experiments are still needed. The experimental data and its results are not used to limit the scope of rights of the present invention.

The aforementioned preferred embodiments are merely examples of the present invention and its technical features, and the technology of this embodiment may still be appropriately implemented in various substantially equivalent modifications and/or replacement methods; therefore, the scope of rights of the present invention shall be subject to the scope defined in the claims of the patent application.

Claims (20)

A method for preparing a Spartina officinalis fermentation complex capable of delaying aging and promoting sleep through brain dopamine production, characterized in that it comprises the following steps: The Junmi grass raw material crushing stage: the Junmi grass raw materials are respectively squeezed to obtain a Junmi grass crude extract, wherein the Junmi grass raw materials are Gastrodia elata fruiting bodies, black rice, and wheat grass; the Junmi grass crude extract is a Gastrodia elata fruiting body crude extract, Gastrodia elata fruiting body residue, black rice crude extract, black rice residue, wheat grass crude extract, and wheat grass residue; Negative pressure wall breaking stage: the crude extract of multiple fungi Spartina officinalis is placed in Next, effect Day 1, obtaining the first extract; Extraction fermentation stage: 0.1-0.5% (w/w) pectin decomposing enzyme and 0.2-2% (w/w) lactic acid bacteria are implanted into the first extract, wherein the extraction fermentation stage is to control the fermentation temperature at The fermentation is continued for 8 to 14 days to obtain a first fermentation liquid; Modification fermentation stage: 0.2-2% (w/w) yeast or acetic acid bacteria is implanted into the first fermentation broth, wherein the fermentation temperature of the modification fermentation stage is controlled at Continuous fermentation Days later, the second fermentation broth was obtained. The preparation method according to claim 1, characterized in that the crushing stage of the rice grass raw material further comprises a hot water treatment step, wherein the gastrodia fruiting body, black rice and wheat grass are Soak in hot water for 30 minutes. The preparation method according to claim 1, characterized in that the negative pressure wall breaking stage further includes an isotonic pressure increasing step, wherein the isotonic pressure increasing step is to add of sugars. The preparation method according to claim 3, characterized in that the sugars include one or more of organic brown sugar, isomalto-oligosaccharides, sorbitol, granulated sugar, and sucrose. The preparation method according to claim 1, characterized in that the lactic acid bacteria include one or more of Lactobacillus plantarum, Lactobacillus delbrueckii, Lactococcus lactis, Lactobacillus acidophilus or B. bifidum. The preparation method according to claim 1, characterized in that the yeast or the acetic acid bacteria comprises one or more of Aspergillus fibuligera, Saccharomyces cerevisiae, Pichia faciens, S. pombe, A. hansenii, A. xylinum or A. suboxydans. The preparation method according to claim 1, further comprising a granulation stage, wherein the second fermentation broth is filtered and granulated by spray drying. A Spartina fermentation complex that can delay aging and soothe sleep through brain dopamine production is obtained according to the preparation method as claimed in claim 1, characterized in that the Spartina fermentation complex comprises the multiple Spartina crude extracts, the lactic acid bacteria, and the yeast or the acetic acid bacteria, wherein the multiple Spartina crude extracts are Gastrodia elata fruiting body crude extract, Gastrodia elata fruiting body residue, black rice crude extract, black rice residue, wheat grass crude extract, and wheat grass residue. A use of the Spartina umbellata fermentation complex obtained according to the preparation method as claimed in claim 1, characterized in that the Spartina umbellata fermentation complex increases the number and activity of lactic acid bacteria that can synthesize γ-aminobutyric acid in the intestine of mammals. The use according to claim 9 is characterized in that it can effectively enhance the synthesis of a γ-aminobutyric acid (GABA) precursor in the intestine of a mammal. The use according to claim 9, characterized in that the γ-aminobutyric acid (GABA) precursor continues to be generated for more than 8 hours. The use according to claim 9, characterized in that the mammal is selected from cats, dogs, rabbits, cows, horses, sheep, goats, monkeys, mice, rats, gerbils, guinea pigs, pigs, or humans. A use of the Spartina officinalis fermentation complex obtained according to the preparation method as claimed in claim 1 for preparing a drug for improving the sleep quality of an individual, wherein the average deep sleep time of the individual can be maintained within a normal range. The eye movement period can be restored and maintained at The effective dosage of the Spartina officinalis fermentation complex is 250 mg/70 kg person/day to 750 mg/70 kg person/day per dose. A use of the Spartina umbellata fermentation complex obtained according to the preparation method as claimed in claim 1 for preparing a drug for preventing Parkinson's disease, characterized in that the effective dose of the Spartina umbellata fermentation complex is 250 mg/70 kg person/day to 750 mg/70 kg person/day per dose. A use of the Spartina umbellata fermentation complex obtained according to the preparation method as claimed in claim 1 for preparing a drug for treating Parkinson's disease, characterized in that the effective dose of the Spartina umbellata fermentation complex is 500 mg/70 kg person/day to 750 mg/70 kg person/day per dose. A use of the Spartina umbellata fermentation complex obtained by the preparation method as claimed in claim 11 for preparing a drug for improving brain antioxidant indicators, characterized in that the antioxidant indicators are SOD activity, G6PD activity and GPx activity, and the effective dose of the Spartina umbellata fermentation complex is 250 mg/70 kg person/day to 750 mg/70 kg person/day per dose. A use of the Spartina umbellata fermentation complex obtained by the preparation method as claimed in claim 1 for preparing a drug for protecting nerves, characterized in that the effective dose of the Spartina umbellata fermentation complex is 250 mg/70 kg person/day to 750 mg/70 kg person/day per dose, wherein the nerve protection includes preventing neuronal damage or preventing substantia nigra damage. A use of the Spartina umbellata fermentation complex obtained by the preparation method as claimed in claim 1 for preparing a drug for treating nerve damage, characterized in that the effective dose of the Spartina umbellata fermentation complex is 500 mg/70 kg person/day to 750 mg/70 kg person/day per dose, wherein the nerve treatment includes treating neuronal damage or treating substantia nigra damage. A use of the Spartina umbellata fermentation complex obtained by the preparation method as claimed in claim 1 for preparing a drug for preventing dopamine reduction in the brain, characterized in that the effective dose of the Spartina umbellata fermentation complex is 250 mg/70 kg person/day to 750 mg/70 kg person/day per dose. A use of the Spartina umbellata fermentation complex obtained according to the preparation method as claimed in claim 1 for preparing a drug for treating decreased dopamine in the brain, wherein the effective dose of the Spartina umbellata fermentation complex is 500 mg/70 kg person/day to 750 mg/70 kg person/day per dose.