TWI407968B - Composition and method for prevention and treatment of alzheimer's disease - Google Patents

Abstract

This invention presented is a composition and method for prevention and treatment of Alzheimer's disease, the composition comprising monacolins and anti-inflammation agents is made by the method of the invention. The composition of the invention has effect on prevention and treatment of Alzheimer's disease without noticeable side effects caused to patients.

Description

Composition for treating and preventing Alzheimer's disease and preparation method thereof

The invention relates to a composition for treating and preventing Alzheimer's disease and a preparation method thereof, in particular to a treatment and prevention of monacolins, anti-inflammatory substances and antioxidant substances from red glutinous rice. The composition of Alzheimer's disease can be applied to consumers by means of tablets, capsules, powders, drinks and the like.

Alzheimer's disease (AD), also known as Alzheimer's disease, is a persistent neurological disorder. People who get Alzheimer's disease will gradually lose their memory and have language and emotional barriers. The gradual loss of intelligence is called dementia. When the disease becomes more and more serious, the patient needs the assistance of others in all aspects of life, such as taking a bath, eating, going to the toilet, and so on. Because people with Alzheimer's disease need day and night care, the lives of their relatives and friends are often greatly affected. Loss of memory may be the most obvious symptom of Alzheimer's disease, especially if you can't remember what happened not recently or recently. The initial symptoms appear subtle and gradual, imperceptible, and these symptoms may also occur in other dementias that are not specific to Alzheimer's disease. For example: Lost in a familiar place, forgetting that something has been done, always revisiting old things or not being able to learn new things. When the condition worsens, the patient may not be able to find the appropriate word or make a major decision during the conversation. One of the most painful aspects of Alzheimer's disease is that patients sometimes have no way to recognize relatives and friends. The patient's personality may also become abnormally irritating, paranoid and disliking, and do not like to interact with people. In the later period, the patient may appear on the street and get lost. New research shows that areas of the brain that deal with visual and spatial information in Alzheimer's patients may be damaged. This explains why patients have no way of recognizing where they are or are confused. Patients may also become unfocused and therefore unable to care for the needs of their own daily body. Other areas of the brain that are important for memory in Alzheimer's disease are also affected, such as the basal forebrain and hippocampus. Many people suffering from Alzheimer's disease die from other causes, such as pneumonia. From the date of diagnosis, patients with Alzheimer's disease generally have a life span of 6-8 years, but many patients still survive for more than 20 years.

There are currently about five drugs for the treatment of Alzheimer's disease on the market that are approved by the US Food and Drug Administration (FDA). These drugs are cholinesterase inhibitors (non-trade name tacrine and donepezil). Cholinesterase is a key enzyme in the decomposition of acetylcholine. These drugs block enzymes by blocking choline esters ( The role of cholinesterase to inhibit the decomposition of acetylcholine. Both drugs can increase the content of acetylcholine in the brain. Both drugs delay the loss of memory and help the patient perform the actions required for daily living. But it is important that these drugs do not cure Alzheimer's disease, they can only alleviate the symptoms of Alzheimer's disease. In addition, these drugs can cause some side effects to patients, such as nausea, headache, diarrhea, insomnia, pain, hallucinations or dizziness, etc., so it is not a practical treatment for Alzheimer's disease.

As mentioned above, such effective treatment and prevention of Alzheimer's disease, without any obvious adverse side effects, is a practical thinking direction.

Accordingly, the main object of the present invention is to provide a composition for treating and preventing Alzheimer's disease for the purpose of effectively treating and preventing Alzheimer's disease.

Another object of the present invention is to provide a composition for treating Alzheimer's disease which is effective in treating and preventing Alzheimer's disease without causing any significant adverse side effects.

A composition for treating and preventing Alzheimer's disease according to the present invention, which is a composition comprising monacolins, an anti-inflammatory substance and an antioxidant substance, which can be applied to a tablet, Capsules, powders, drinks, etc. are available to consumers. The composition formed by the three kinds of substances is used for treating and preventing Alzheimer's disease. In addition, in the practice of the present invention, the minimum effective amount of monacolins is at least more than 100 μg, and the effective minimum content of anti-inflammatory substances is at least More than 40μg, and the effective minimum content of antioxidants is at least greater than 10μg, and the optimal weight ratio of Monacolins, inflammatory substances and antioxidants is 40:2:1 to achieve the treatment and Prevention and efficacy without any obvious adverse side effects.

Furthermore, a composition for treating and preventing Alzheimer's disease disclosed in the present invention is a composition comprising monacolins and an anti-inflammatory substance, which is effective by Monacolins. The minimum content must be at least 200 μg, and the effective minimum content of anti-inflammatory substances should be at least greater than 60 μg, and the optimal weight ratio of Monacolins to inflammatory substances is 10:1, and the treatment can be achieved in this composition. It has the effect of preventing Alzheimer's disease and not causing any obvious adverse side effects.

Furthermore, in the present invention, a method for preparing and preventing an Alzheimer's disease composition is as follows: firstly, one meter is cleaned and sterilized under a high pressure and high temperature environment, and then a specific red peony strain is inoculated to In a culture medium and in a first specific temperature and humidity environment, a specific oscillating environment and a first specific time limit for inoculation, after the completion of the sterilization, stirring the inoculum culture medium in a first specific temperature environment and continuously providing for a second specific time limit a specific proportion of moisture is appropriately stirred at a fixed time in the first specific temperature environment for a post-ripening within a third specific time limit, and the red mash fermentation product after completion of the fermentation is collected and maintained for the fourth specific period. Drying at a second specific temperature, grinding the dried red peony fermentate into a powder and analyzing whether it meets the one-component ratio condition, when the composition for treating and preventing Alzheimer's disease according to the present invention is The prepared powder of the red sorghum fermented product is added to the edible beverage in a specific ratio to prepare a treatment and prevention of Alzheimer's The red glutinous beverage can also be filled into a capsule or prepared into a tablet by preparing the powder of the red sorghum fermented product. In this way, the composition can be implemented and the treatment and prevention of Alzheimer can be achieved. And the effect of not causing any obvious adverse side effects.

The details and techniques of the present invention are described below:

The present invention is a composition for treating and preventing Alzheimer's disease, and the concept of the present invention will be described before introducing the composition of the present invention. The main factor in the formation of Alzheimer's disease is the deposition of amyloid β-protein (Aβ) in the brain, and the lack of neurotransmitters and oxidative inflammation cause the disease to gradually worsen. The production of Aβ is a fragment protein produced by cleavage of a secretase-like amyloid precursor protein (APP) by a secretase. The sheared enzymes contain alpha, beta and gamma-secretase. Aβ is a β-secretase that can cleave the amino acid linkages of 671 and 672, and γ-secretase can cleave Aβ fragments on the APP near 713. When the enzyme cleaves to the amino acid sequence, the remaining fragments are produced in addition to the 1-40, 1-42 fragments. If the APP is cleaved by α-secretase to produce a fragment of sAPP-α and p10 kDa, p10 can be cleaved by γ-secretase to generate p 3 and p 7 fragments, wherein p3 is a small fragment of Aβ. In addition, if APP alone is cleaved by β-secretase to produce fragments of sAPP-β and p12 kDa, the p12 fragment can be cleaved again by γ-secretase to produce Aβ and p7 fragments (Evin et al. 2003; Shoji et al. 1992). Although these small fragments have no ability to aggregate in the brain, they can demonstrate the cleavage activity of different types of secretase. When APP is decomposed by β-secretase and γ-secretase, various types of Aβ are produced. Among them, soluble Aβ is not toxic, and it must be polymerized to form fibrils, which is toxic to nerve cells. Soluble Aβ is not toxic and must be polymerized to form Aβ fibrils before it can be sterilized by disrupting intracellular calcium ion balance and inducing oxidative stress. Aβ polymers can also interfere with axonal elongation by promoting hyperphosphorylation of intracellular tau, resulting in cell death. In addition, Aβ fiber bundles can be activated by binding to specific receptors on the microglia and astrocyte membranes in the brain, which release a variety of neurotoxins, including interleukin-1 (IL-1) and interleukin- 6 (IL-6), tumor necrosis factor (TNF-α) and other inflammatory substances and free radicals such as nitric oxide and superoxide anion attack the nerve cells, causing their death. In recent years, it has been found that the apoE (apoE, apolipoprotein E) gene on the 19th pair of chromosomes is associated with Alzheimer's disease. The expression of ApoE and Aβ is rapidly combined and deposited in the brain to form plaques of the elderly, which will cause nerve cells. Necrosis.

In addition, many literatures indicate that cardiovascular disease has a certain correlation with Alzheimer's disease. According to Martha et al., the elderly who are over 65 years old are surveyed. The elderly who are rich in saturated fatty acids that cause cardiovascular disease in their diets have been investigated. After 4 years, most elderly people will suffer from Alzheimer's disease. In contrast, the diet rich in polyunsaturated fatty acids is inversely related to the elderly with monounsaturated fatty acids (Freund-Levi et al. 2006). There are many studies demonstrating that the production of Aβ that causes Alzheimer's disease is associated with intracellular lipid metabolism (Frears et al. 1999; Kuo et al. 1998; Roher and Kuo 1999). In recent years, studies have shown that Aβ is altered by intracellular cholesterol distribution and cholesterol esterification (Frears et al. 1999). ApoE promotes the increase of cholesterol in plasma and is one of the risk factors for cardiovascular disease. Puglielli et al. (2001) indicated that intracellular acetylcholine activity in hamsters was positively correlated with cholesterol lipid content (Puglielli et al. 2001; Zhao et al. 2005). Therefore, cardiovascular disease has a positive correlation with Alzheimer's disease.

Thus, the compositions of the present invention focus on reducing the deposition of A[beta] in the brain and reducing saturated fatty acids that cause cardiovascular disease for the treatment of Alzheimer's disease.

In recent years, related research on Monascus species has been increasing. In the future development, Hung Hom will be developed into a multi-functional health food. The monacolin K, γ-aminobutyric acid (GABA) and antioxidant substances in red peony have been confirmed by the present invention to greatly improve the feasibility of improving Alzheimer's disease for treatment and prevention.

Monacolin K's alias contains lovastatin, a rate-limiting enzyme for cholesterol synthesis - 3-hydroxy-3-methylglutaryl-coenzyme A reductase, an inhibitor of HMG CoA reductase, is a statin-like compound that has been clinically proven to be effective in improving cardiovascular disease (CAD). Epidemiological studies in recent years have also pointed out that patients with Alzheimer's disease treated with statin do have a good improvement effect. In the UK General Practitioners Research Database, 70% of patients with Alzheimer's disease treated with statin had significant improvement (Jick et al. 2000); similar experiments still included 2000. In the year, Wolozin et al. used Alzheimer's disease in the elderly in three different hospitals in the United States as a subject. The diagnosis was based on NINCDS-ADRDA (National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related). According to the Disorders Association, the subjects were divided into three groups, group A was the control group, group B was treated with statins, and group C was treated with other cardiovascular diseases (including blood pressure lowering drugs). The result is surprisingly that when statins are used to treat patients, the degree of Alzheimer's disease can be reduced by 70%, with lovastatin and palavastatin (pravastatin). ) works best (Wolozin et al. 2000).

Nicotinic, muscarinic, serotonin, glutamate receptors and gamma-aminobutyric acid (γ-aminobutyric acid) in the brains of Alzheimer's patients GABA), norepinephrine and other nerve impulse transfer factors are all defective. The brain will be reduced by 10% during aging, mainly due to the decrease in the number of neurons in the cerebral cortex, which may even decrease by 30-50% in some areas, and the neurotransmitters will also decrease, such as acetylcholine Alkali, GABA, catecholamine, and the like. Due to the reduction of these substances, the cognitive function of the brain is reduced or the memory ability is impaired. The earliest symptoms are memory loss, loss of ability to judge things, and easy to get lost. In the advanced stage, the patient may have language disorders, abnormal behavior, paralysis, and life cannot take care of themselves (Mohr et al. 1994).

When the granular gland is deficient in O ₂ or H ₂ O ₂ and OH‧, it leads to the formation of highly reactive OH‧. The release of Aβ by free radicals leads to the formation of insoluble Aβ, which worsens Alzheimer's disease (Hsieh and Tai 2003). Experiments have found that oral antioxidants such as vitamin C and vitamin E combined preparations can effectively alleviate the symptoms caused by Alzheimer's disease (Yallampalli et al. 1998; Yamada et al. 1999). In addition, it is called ceramlde, a natural nerve sheath that counteracts the effects of Aβ and FeSO _{4 on} hippocampal nerve injury (Abousalham et al. 2002). The antioxidant capacity of red peony was proposed by Aniya et al. in 1999. The antioxidant mechanism of dimerumic acid in red carp can be found to remove ‧ OH, ‧ O ₂ ^- , ferryl-Mb, peroxyl The radicals and the inhibition of lipid peroxidation (LPO), which provide an electron to the oxide to oxidize itself to a nitroxide radical, and the nitroxide radical is removed again. Lose to achieve antioxidant effects (Taira et al. 2002). The anti-oxidant effect of red peony can be used in the prevention of Alzheimer's disease to avoid the disease being aggravated by the attack of oxidative free radicals.

The present invention was published on July 30, 2007 in the Journal of Neuroscience Research ( Lee CL , Kuo TF, Wang JJ, Pan TM: Red mold rice ameliorates impairment of memory and learning ability in intracerebroventricular amyloid beta-infused rat via repressing amyloid J Neurosci Res. 2007, 85, 3171-7182.) The animal used in the study of the present invention is a Wistar strain male rat purchased from the Experimental Animal Center of the National Taiwan University School of Medicine, 8 weeks old, weighing About 250 g, 7 in each group, kept in an air-conditioned space, the temperature is maintained at 23±1 °C, the lighting control is 12 hours, 12 hours, dark (08:00 light, 20:00 light), food and water are not given limit. Rats were pre-cultured to a body weight of 300 g. The experimental group and brain infusion were implanted. The daily intake of red glutinous rice and the amount of Aβ40 injected in each group are shown in Table 3-2. The Vehicle group was a rat who underwent surgery but did not infuse Aβ40 as a normal memory learning ability. The Aβ group was an Alzheimer's disease rat who underwent surgical infusion of Aβ40 as a memory learning dysfunction. The feeding dose of red glutinous rice was via FDA. The published Bodyd's Formula of Body Surface Area is calculated (Boyd 1935; Lee et al. 2006). One of the red glutinous rice feeding doses in this study was equivalent to 2 g of red glutinous rice per day for adults (170 cm, 65 kg), high dose red glutinous rice for RH group, and daily feeding for LS group. The same dose of lovastatin (1.43 mg/kg rat) as the RL group was used to compare the treatment of Alzheimer's disease with simple monacolin K and red scorpion containing monacolin, antioxidant and anti-inflammatory substances. And the effect of prevention.

Alzheimer's disease rats were infused with Aβ40 for 28 consecutive days in the rat brain, causing Aβ40 to deposit directly in the brain and causing Alzheimer's disease. During the experiment, the red sorghum fermentation product or the same concentration of lovastatin was fed during the experiment. To investigate the inhibitory effect of red peony on the production of Aβ toxicity and to evaluate the impact on memory learning ability. The experimental results of the present invention show that infusion of Aβ40 in the brain of rats for 28 consecutive days increases the activity of acetylcholine activity, the amount of reactive oxygen species (ROS) and lipid peroxidation in the brain, and Reduce total antioxidant capacity and superoxide dismutase (SOD) activity. The feeding of red glutinous rice can significantly inhibit the damage caused by infusion of Aβ40 in the brain, and the effect is better than that of the lovastatin drug treatment group. In addition, the infused Aβ40 cannot be deposited in a large amount in the hippocampus back tissue by inhibiting the oxidative inflammatory response. Red glutinous rice was first confirmed to have a lower risk factor for Alzheimer's disease and improved memory learning ability, and its effect is more significant than lovastatin drugs.

The test grouping of the present invention is shown in Table 1:

The test apparatus of the present invention is as shown in Fig. 1. The test apparatus 10 includes a bright room 11 and a dark room 12 of the same size, and a 10 (W) × 10 (D) cm gate 13 is provided in the center of the two chambers, when the gate 13 is opened. The bright room 11 and the dark room 12 can be connected. The bright room 11 has illumination, while the dark room 12 has no light, and the bottom is provided with a metal wire 14 arranged in parallel at intervals of 1 cm, and is supplied with current. The experimental procedure is to first place the rats of each group in the bright room 11 and simultaneously open the gate 13. Immediately after the rat entered the dark room 12, the gate 13 was closed, and current (100 V, 0.3 mA, 2 sec) was applied to the wire 14, and 5 seconds after the electrical stimulation, the rat was taken out from the dark room and returned to the cage. If the rat does not enter the darkroom 12 after 300 seconds, the rat is forced into the darkroom 12 and the gate 13 is closed, and the current of the metal wire 14 at the bottom of the darkroom 12 is returned to the home cage. Measurement period: At 24 hours and 48 hours after the training, the rats were placed in the bright room 11 while the gate 13 was opened, and the step-through latency of the rats in the bright room 11 was recorded. When the residence time in the bright room 11 is more than 300 seconds, the learning and memory ability of the rat is said to be normal.

The time that the rat stays in the bright room 11 until entering the dark room 12 can be regarded as an indicator for evaluating the memory learning ability in the passive avoidance test. As shown in Fig. 2, all the rats immediately entered the dark room 12 during the first test due to the darkening characteristics, and the electric shock stimulated in the dark room 12 caused the rats with normal memory learning ability to be next. During the test, I was afraid to enter the darkroom 12 again. Therefore, in the second trial, the time that each group of experimental animals stayed in the bright room 11 will be significantly different due to the difference in memory learning ability. The results of the second trial showed that the Aβ group stayed in the bright room for significantly less time than the vehicle group. The residence time of the experimental group RL and RH in the bright room was significantly longer than that of the Aβ group ( p <0.05). Although the effect of the Lovastatin group was not as good as the RL group with the same monacolin K content, the effect of memory learning was still significantly higher than that of the Aβ group.

Another test device of the present invention, as shown in Fig. 3, is a water labyrinth device 20 having a circular swimming pool 21 having a diameter of 140 cm and a height of 45 cm. The swimming pool contains a movable rest platform P1 (or Called escape platform, escape platform). The platform has a diameter of 12 cm and a height of 25 cm. Before the experiment, the circular swimming pool 21 must be filled with water to a liquid level of 27 cm. The circular pool 21 is divided into four quadrants (I, II, III and IV zones) and five starting points are set, and the rest platform P1 is placed at the center of either quadrant. During the test, a camera was placed directly above the center point of the pool to record the swimming path of the experimental animals.

The effect of red glutinous rice on the impairment of reference memory learning ability in Alzheimer's disease rats with Aβ transfused into the brain is shown in Fig. 4. The time from the start of the water maze device 20 to the finding of the resting platform P1 of the experimental animals is regarded as an indicator for evaluating the reference memory test. From the results of the second to the ninth memory training, the Alzheimer's disease rats with Aβ infusion always took a longer time to find the rest platform P1 than the normal memory group, but fed the red scorpion. Alzheimer's disease rats (RL and RH groups) significantly reduced the time of search ( p < 0.05). Although the Lovastatin diet can take a shorter search time than the Aβ group, its effect is lower than that of the RL group and the RH group. The same search time will present a similar swim path length. In addition, there was no significant difference in swimming speed between the groups of experimental animals (data not shown) ( p > 0.05).

The spatial detection test was followed by the last reference memory test on day 24. The rest platform P1 will be removed from the water maze device 20 after the end of the reference memory test, and the time when the experimental animal is placed in the quadrant of the resting platform P1 is regarded as an indicator of the memory learning ability of the spatial detection test. As a result, as shown in Fig. 4A, the time required for the Aβ group to search in the target quadrant was significantly lower than that of the vehicle group ( p < 0.05). In the RL group and the RH group, the search time in the target quadrant was 38.2% ( p < 0.05) and 48.0% ( p < 0.01), respectively, which proved that red glutinous rice can improve memory learning in Alzheimer's rats. Impaired ability. The swimming path can help to judge the authenticity of the memory learning ability of experimental animals in the spatial detection test. Figure 4B shows that the Aβ group has no directional and non-targeted target quadrants in the spatial detection test, and the swimming path extends throughout the circular pool 21. Conversely, the RL group, the RH group, and the vehicle group with better memory learning ability will directly swim into the target quadrant, and it takes a long time to hang in the target area. However, the lovastatin diet group still showed an improvement in memory learning ability between the RL group and the Aβ group (Fig. 4B and Fig. 4C). The working memory test is a kind of short-term memory learning ability evaluation method. The results are shown in Fig. 5. The time of finding the resting platform P1 in the Aβ group is significantly longer than that in the vehicle group ( p< 0.01). However, the RL group and the RH group exhibited memory learning effects in the working memory test as in the normal memory group, and were shortened by 57.3% and 58.9% compared with the Aβ group ( p < 0.01). Although the LS group was significantly shorter than the Aβ group by 26.7% of the search time ( p < 0.05), the effect was lower in the RL group and the RH group ( p < 0.05).

Aβ has been shown to be a major risk factor for memory learning in patients with Alzheimer's disease (Hashimoto et al. 2005; Stephan and Phillips 2005). The manner in which Aβ40 is infused into the lateral ventricles of the rat brain has been shown to successfully establish an animal model of Alzheimer's disease (Stephan and Phillips 2005). The model animals established in this way also have the characteristics of insufficient memory behavior and learning ability (Kar et al. 1998; Schubert et al. 1995; Townsend and Pratico 2005). Infusion of Aβ induces free radicals and reactive oxygen atoms and causes neuronal damage and synaptic elongation to be inhibited, resulting in a lack of memory learning ability (Townsend and Pratico 2005). Therefore, there are several studies that have used antioxidant and anti-inflammatory substances to reduce the risk of oxidative inflammation in the brain of these Alzheimer's diseases and to reduce the ability to learn memory (Chauhan et al. 2004; Cordle et al. 2005).

The Morris water maze test is one of the main methods for assessing memory learning ability. The reference memory test represents a long-term memory ability evaluation method, while the working memory test is a short-term memory ability evaluation method. According to the results of the above memory learning test, Alzheimer's rats fed a single dose and five times the dose of red glutinous rice can shorten the search time of the reference memory test and the working memory test ( p < 0.05). In addition, the red glutinous rice diet group can extend the search time of the target quadrant in the spatial detection test. These results clearly indicate that feeding red glutinous rice can improve memory learning ability and find a resting platform for the water maze in a shorter time. Conversely, untreated Alzheimer's rats searched the entire water maze in an untargeted manner, so it took more time in both the reference memory and the working memory test.

Monacolin K is one of the main functional components of red glutinous rice to lower blood fat, and statin compound has been considered to treat Alzheimer's disease because it has the effect of inhibiting the formation and anti-inflammatory of Aβ in the brain. Emerging drugs (Chauhan et al. 2004; Li et al. 2006). Most Alzheimer's disease treatment studies use statins to reduce the production of Aβ in the brain of mice transgenic with Alzheimer's disease (Yamada et al. 1999). In the current known studies, lovastatin has not been used to alleviate the impairment of memory learning ability of Alzheimer's disease model rats with Aβ infusion. In this study, lovastatin was used to replace monacolin K in red glutinous rice to explore whether monacolin K in red glutinous rice is the only substance that can alleviate the memory learning ability of Alzheimer's disease. However, the results of the above memory learning experiments indicated that the lovastatin feeding group was less effective in reducing memory learning impairment than the RL group with the same monacolin K concentration. Therefore, it was confirmed that monacolin K in red glutinous rice is not the only functional component that alleviates the damage of memory loss caused by Aβ.

Monacolin K is one of the main functional components of red glutinous rice, and statin has been considered as an emerging drug for the treatment of Alzheimer's disease because it has the effect of inhibiting the production and anti-inflammatory of Aβ in the brain (Chauhan et al) 2004; Li et al. 2006). Most Alzheimer's disease treatment studies use statins to reduce the production of Aβ in the brain of mice transgenic with Alzheimer's disease (Yamada et al. 1999). In the current known studies, lovastatin has not been used to alleviate the impairment of memory learning ability of Alzheimer's disease model rats with Aβ infusion. In this study, lovastatin was used to replace monacolin K in red glutinous rice to explore whether monacolin K in red glutinous rice is the only substance that can alleviate the memory learning ability of Alzheimer's disease. However, the results of the above memory learning experiments indicated that the lovastatin feeding group was less effective in reducing memory learning impairment than the RL group with the same monacolin K concentration. Therefore, consumption of monacolin K can effectively improve the symptoms of Alzheimer's disease caused by Aβ.

Current treatments for Alzheimer's disease focus on inhibiting acetylcholine activity to increase acetylcholine concentration and improve cognitive and memory behavior (Nabeshima and Nitta 1994). Related studies established in Alzheimer's disease model animals have also demonstrated that continuous infusion of Aβ into the rat brain will result in neurotoxicity of choline neuronal decline and deficiency. Studies have also shown that the brain acetylcholine content in the brain of rats with Aβ Alzheimer's disease is significantly less than that of normal rats (Arendt et al. 1984; Darvesh et al. 2004). A decrease in acetylcholine concentration and an increase in acetylcholine activity will exacerbate neuronal loss (Stephan and Phillips 2005). In addition, increased acetylcholine activity has been noted to cause a greater degree of polymerization of Aβ and to form a more stable and more toxic fibrillar amyloid (Aβ fibrils) (Stephan and Phillips 2005). Therefore, inhibition of the activity of acetylcholine enzyme is considered to be an indirect reduction of the neuroprotective effect of Aβ-induced memory learning. Figure 6 shows the effect of red glutinous rice on the activity of acetylcholine in the cerebral cortex and hippocampus of rats with Aβ40 infusion. The figure indicates that the acetylcholine activity of the cerebral cortex of the Aβ group is higher than that of the vehicle group. Increased by 50.5%, and increased by 179.1% in the hippocampus. Both doubling and five-fold doses of red glutinous rice significantly inhibited the activity of acetylcholine enzyme increased by Aβ. However, lovastatin had no inhibitory effect on acetylcholine activity in cerebral cortex and hippocampus. This result confirmed that the red glutinous rice has a substance that inhibits the activity of acetylcholine enzyme, and this substance should not be monacolin K but other functional metabolites. Little research indicates that lovastatin inhibits the activity of acetylcholine enzyme. Studies have shown that lovastatin does not have a significant inhibitory effect on the activity of acetylcholine. The results of this study are consistent with the trends of the present invention. In addition to monacolin K, the complex efficacious metabolite of red glutinous rice may contain other acetylcholine inhibitors. However, GABA, a metabolite in red glutinous rice, is also a neurotransmitter that also helps improve the cognitive and memory behavior of Alzheimer's disease.

Aβ has been shown to cause oxidative stress in the brain of Alzheimer's patients. Therefore, inhibition of Aβ-induced oxidative stress is considered to be an important indicator for the development of Alzheimer's disease drugs. As shown in Figure 7, infusion of Aβ into the brain resulted in a decrease in total antioxidants status (TAS) in the cerebral cortex and hippocampus tissue by 20.9% and 20.4%, respectively, compared to the vehicle group. Feeding lovastatin only increased the TAS of the cerebral cortex by 13.9%, but it was not effective in hippocampal gyrus. The TAS in the cerebral cortex and hippocampus of the RL group was significantly increased by 24.6% and 46.2%. However, this study found that TAS in the cerebral cortex and hippocampus of rats fed Alzheimer's disease with a five-fold dose of red glutinous rice was significantly higher than that of the Aβ group.

The effect of red glutinous rice on the concentration of malondialdehyde (MDA) in the cerebral cortex and hippocampal gyrus, as shown in Figure 8. MDA concentrations in the cerebral cortex and hippocampus were significantly increased by 95.3% and 112%, respectively, after transfusion of Aβ in the brain, but this increase in MDA concentration was effectively reduced again by increasing the feeding dose of red mites.

The superoxide dismutase (SOD) activity in the cerebral cortex and hippocampus of the RL group and the RH group also showed similar neuroprotective effects, as shown in Fig. 9. Although SOD activity in the cerebral cortex and hippocampus was reduced by 19.8% and 25.2%, respectively, in the brain, a double dose of red glutinous rice would increase the hippocampus in the cerebral cortex and hippocampus. SOD activity was 27.2% and 52.7%, and 5 times of red glutinous rice increased by 27.2% and 60.9%, respectively.

From the above results, it is known that Alzheimer's disease model rats with impaired memory learning have severe oxidative stress in the cerebral cortex and hippocampus. However, these oxidative stresses are improved by daily feeding of red glutinous rice, and the effects are not only dose-effect, but also better than the lovastatin feeding group. Antioxidant substances in the fermentation products of red peony have been mentioned as dimerumic acid, tannin, phenol, monounsaturated fatty acid, superoxide. Disproportionation enzymes and sterols (Aniya et al. 1999; Wang et al. 2006).

Therefore, the antioxidant substance of the present invention can be derived from dimerumic acid, tannin, phenol, monounsaturated fatty acid, superoxide dismutase and sterol. (sterols) and any of superoxide dismutase (SOD).

The results of the in vivo animal test of the present invention are shown in Fig. 10A. The brain infusion of Aβ significantly increases the concentration of reactive oxygen species (ROS) in the cerebral cortex and hippocampus of the rat, respectively, by 39.8%. 28.7% ( p < 0.05). The concentration of ROS in the cerebral cortex and hippocampus of rats with Alzheimer's disease fed daily red glutinous rice decreased. The RL group was reduced by 16.0% ( p < 0.05) and 21.2% ( p < 0.05), respectively, and the RH group was decreased by 35.4% ( p < 0.01) and 21.3% ( p < 0.05), respectively. The Lovastatin diet reduced the ROS concentration in the cerebral cortex by 29.3% and decreased by 15.7% in the hippocampus ( p < 0.05). The expression of inducible nitric oxide synthase (iNOS) in hippocampus is shown in immunohistochemical staining in Figure 10B. Aβ infusion significantly increased iNOS expression in hippocampal gyrus of RL and RH groups. It was found that the amount of expression of iNOS was significantly suppressed. However, it was also found that the anti-inflammatory effect of the lovastatin feeding group was worse than the 1x and 5x doses of the red scorpion feeding group.

The concentration of ROS in the cerebral cortex and hippocampal gyrus of Alzheimer's disease rats was significantly reduced by increasing the dose of red glutinous rice. Although lovastatin has been shown to be effective in inhibiting the inflammatory response induced by Aβ in cell mode, such effects have not been demonstrated in animal models, and the anti-inflammatory effects of statins have not been applied to the reduction of brain transfusion Aβ. Injury in memory learning in rats with heather. This study confirmed that lovastatin feeding significantly reduced ROS content and iNOS expression in brain tissue, but these effects were slightly worse than the red glutinous rice diet group. In recent years, the health benefits of secondary metabolites of red peony have been gradually developed. Therefore, many anti-inflammatory substances have been proposed, including various forms of monacolins and 6 kinds of red peony metabolites (azaphilones). ): monascin, ankaflavin, rubropunctatin, monascorburin, rubropunctamine and monazkamine (monascorburamine), 2 species of furanoisophthalides: xanthomonasin A and xanthomonasin B; and 2 amino acids ): (+)-monascus acid ((+)-monascumic acid) and (-)-monascumic acid (Schubert et al. 1995). Therefore, monacolin K is not the only metabolite that inhibits the inflammatory response caused by Aβ. Akihisa et al. (2005) used an anti-inflammatory response caused by 12-O-tetradecanoylphorbol-13-acetate (TPA) in a mouse model. And confirmed that the main anti-inflammatory substances are azaphilones and furanoisophthalides. Based on the results of in vitro and in vivo animal evaluation of this study and the anti-inflammatory effects of red peony, the inhibition of Aβ-induced inflammatory response by red glutinous rice is mainly due to the synergistic mechanism of monacolin K and other anti-inflammatory substances.

Therefore, the inflammatory substances mentioned in the present invention can be derived from γ-aminobutyric acid (GABA), monascin, ankaflavin, and rubropunctatin. , monascorburin, rubropunctamine, monascorburamine, xanthomonasin A, and xanthomonasin B And (+)-monascus acid ((+)-monascumic acid) and (-)-monascusic acid ((-)-monascumic acid).

After continuous infusion of Aβ40 to the hippocampal gyrus of the rats for 28 days, it was found from the comparison with the vehicle group in Fig. 11 that a large amount of Aβ40 was deposited in the hippocampal gyrus of the Aβ group. From the above results and previous studies, it can be known that the infused Aβ40 causes oxidative stress and inflammatory reaction in the brain, thereby causing deposition of Aβ40. When the deposited Aβ40 content is more, it will cause more severe oxidative stress and inflammatory reaction, and repeated vicious cycles will cause the brain damage to continue to increase. Lovastatin has the effect of inhibiting the inflammatory response caused by Aβ40, but its ability to resist oxidative stress is weak. Therefore, it can be found that the Aβ40 in the hippocampal gyrus of the LS group is slightly lower than that of the Aβ group, but a large amount of Aβ40 deposition can still be found. The RL group and the RH group fed red glutinous rice had the effect of significantly reducing the accumulation of Aβ40. The reason why red glutinous rice reduces the accumulation of Aβ40 in hippocampal gyrus is mainly due to its inhibition of oxidative stress and inflammatory reaction. Aβ40 infused into the brain is deposited without being promoted by oxidative inflammatory substances. Aβ40 can not cause damage to the brain, and thus effectively improve the ability of memory learning.

The combination of Monacolins, anti-inflammatory substances and antioxidant substances of the present invention for treating and preventing Alzheimer's disease can provide symptoms for improving Alzheimer's disease in red glutinous rice, wherein In one embodiment, the effective minimum content of monacolins per gram of red glutinous rice is at least greater than 100 μg, and the effective minimum content of antioxidants per gram of red glutinous rice is at least greater than 40 μg, and each gram The anti-inflammatory substance in red glutinous rice has an effective minimum content of at least 10 μg; wherein the optimal weight ratio of Monacolins, inflammatory substances and antioxidant substances is 40:2:1, which is the best in the first embodiment. Efficacy, in addition to the effective weight ratio of Monacolins, inflammatory substances and antioxidants ranging from 10:4:1 to 90:2:1 can be applied and applied in tablets, capsules, powders , drinks and other means for consumers to take.

Further, in the second embodiment, a composition of Monacolins and an anti-inflammatory substance can also be used for the treatment and prevention of Alzheimer's disease, when each gram of red glutinous rice is monacolin ( Monacolins) may also have the effect of the above process in compositions having an effective minimum level of at least greater than 200 μg and an effective minimum level of anti-inflammatory substances of at least 60 μg per gram of red glutinous rice; wherein monacolins and inflammation are present. The optimum weight ratio of the substance is 10:1 for the best effect in the second embodiment, and the effective weight ratio of Monacolins to inflammatory substances ranges from 10:3 to 45:1. The second embodiment is carried out and applied to the consumer in the form of tablets, capsules, powders, drinks, and the like.

The anti-inflammatory substance contained in red glutinous rice is γ-aminobutyric acid (GABA), and red glutinous rice is a natural food, which can be fermented and refined in a specific way, so that it is applied to the human body. Does not produce any significant adverse side effects. Therefore, if red glutinous rice is used as a conditioning food for the treatment of Alzheimer's disease, the effect is not only significant, but there is no general medicine that will cause a variety of side effects for patients, which is an invention for the benefit of the population.

Please refer to Figures 12-1 to 12-3 for the preparation and treatment of Alzheimer's disease composition according to the present invention, as follows: First, first clean one meter and in a high pressure and high temperature environment. Sterilization (step 100), this one meter is a rice (Oryza satiVa L., Japonica) , this high pressure high temperature environment refers to a pressure environment of 121 ° C and 1 kg / cm ² , and then a specific The red peony strain is inoculated into a bacterium culture medium and is inoculated in a first specific temperature and humidity environment, a specific oscillating environment and a first specific time limit (step 110), and the bacterium culture medium is composed of at least 5 g of the rice immersion. In 100 ml of distilled sterile water, this first specific time limit means that after 48 hours, the first specific temperature is maintained at 30 ° C, and the specific oscillating environment is maintained at 100 to 150 revolutions per minute (revolutions per minute; Rpm); that is, the red peony strain was inoculated to the inoculum culture medium, and cultured at 30 ° C, shaking at 125 rpm for about 48 hours, and then the bacteria were collected, and 500 g of the solid medium was weighed and washed for 6-8 hours. After filtering the water with cotton cloth, place it on a crepe cloth, flatten it in a tray and sterilize (121 ° C, 20~25 min). Sprinkle water 100 mL. A second sterilization (121 ° C, 20 min) was carried out and the cells were sterilized after cooling. The inoculum was taken into a solid matrix (5%) and stirred well. And then stirring the inoculum culture medium in the first specific temperature environment and continuously providing a specific proportion of water for a second specific time period (step 120), wherein the second specific time limit means within 72 hours, The specific proportion of water refers to 20% distilled sterile water, the culture medium is stirred every 24 hours within 72 hours and supplemented with 20% distilled water and cultured in a 30 ° C incubator, then in a third Within a certain time limit, appropriate agitation is performed at every fixed time in the first specific temperature environment to achieve post-ripening (step 130), and the post-mature refers to a stage in which metabolites are generated, and this third specific time limit is referred to in 96. Within hours. This fixed time means every 10 hours. Within 96 hours, the inoculum culture medium is appropriately stirred every 10 hours, and then one of the red sorghum fermented materials after the completion of the fermentation is collected and dried at a second specific temperature for a fourth specific period. (Step 140), this fourth specific period means that within 24 hours, this second specific temperature means that it is maintained at 55-60 °C. This step is a collection process, and the fermented red sorghum fermented product is subjected to a drying process of 24 hours at 60° C. in an oven, and then the dried red sorghum fermented product is ground into a powder and analyzed for compliance with a component ratio. Condition (step 150), the composition ratio condition means that the composition comprises a monacolins, an anti-inflammatory substance and an antioxidant substance, and the effective weight ratio of the three substances ranges from 10:4:1. To 90:2:1, in this range, it is confirmed that it is in the condition of the ratio of the component. If not, the process is terminated because it is a failed production process; if it is confirmed in the range, step A is further Step B, please refer to Figure 12-2 and Figure 12-3. After Step A, the prepared powder of the red sorghum fermented product is added into the edible beverage in a specific ratio to prepare a treatment and prevention. The sputum drink of Zhaimer's disease (step 160), and complete the production process. This specific ratio is between 1.0 and 4.0%.

After step B, the prepared powder of the red sorghum fermented product is filled into a capsule or made into a tablet (step 170), and the production process is completed.

The production method of the present embodiment can be cultured through a traditional tray, and a smaller type of tray can be made, which is about 20×30×5 cm in length, width and height. When the substrate is cultured, the enamel cloth is wrapped with a solid matrix, which is favorable for turning over. At the same time, the humidity can be maintained; the upper layer is covered with a layer of crepe to isolate the outside pollution, and at the same time maintain the humidity of the red glutinous rice during the fermentation process. The entire culture process is carried out in an open space, and the composition of the present invention can be carried out by the present production method, and can be applied to tablets, capsules, powders, drinks, and the like.

Although the present invention has been described above in terms of the preferred embodiments thereof, it is not intended to limit the present invention, and those skilled in the art may make some modifications and refinements, such as others, without departing from the spirit and scope of the invention. Any article that treats Alzheimer's disease with monacolins, anti-inflammatory substances, and antioxidants is used, and the scope of the present invention is defined by the scope of the appended claims.

10. . . Test device

11. . . Ming room

12. . . darkroom

13. . . Gate

14. . . metal wires

20. . . Water maze device

twenty one. . . Round pool

P1. . . Rest platform

Step 100 Clean one meter and sterilize in a high pressure and high temperature environment

Step 110: inoculating a specific red peony strain into a culture medium and inoculation in a first specific temperature and humidity environment, a specific oscillating environment and a first specific time limit

Step 120: stirring the inoculum culture medium in the first specific temperature environment and continuously providing a specific proportion of moisture within a second specific time limit

Step 130: performing a suitable agitation at a fixed time in the first specific temperature environment within a third specific time limit to achieve the post-ripening

Step 140: Receiving one of the red mash fermentation products after completion of the fermentation and drying it at a second specific temperature for a fourth specific period of time

Step 150: grinding the dried red sorghum fermented product into a powder and analyzing whether it meets the ratio of one component

Step 160: The prepared powder of the red sorghum fermented product is added into an edible beverage in a specific ratio to prepare a red scorpion drink having the treatment and prevention of Alzheimer's disease.

Step 170: filling the prepared powder of the red sorghum fermented product into a capsule or making a tablet

Figure 1 is a schematic view of the test device of the present invention;

Figure 2 is a time chart of the rats staying in the bright room until entering the darkroom;

Figure 3 is a schematic view of the water maze device and sub-area of the present invention;

Figure 4 is a graph showing the effect of red glutinous rice on memory learning ability in a reference memory test and a probe test in rats with amyloid 40 (Aβ40) brain infusion;

Figure 5 is the memory learning ability of red glutinous rice in rats with Aβ40 infusion in the working memory test;

Figure 6 is a graph showing the effect of red glutinous rice on the activity of acetylcholine in the cerebral cortex and hippocampus of rats with Aβ40 infused into the brain;

Figure 7 is a comparison of total antioxidant status (TAS) in the cerebral cortex and hippocampal gyrus of rats with Aβ transfused;

Figure 8 is a graph showing the effect of red glutinous rice on the concentration of malondialdehyde (MDA) in the cerebral cortex and hippocampal gyrus of rats;

Figure 9 is a graph showing the effect of red glutinous rice on the activity of superoxide dismutase (SOD) in rat cerebral cortex and hippocampus.

Figure 10 is the effect of red glutinous rice on the content of reactive oxygen species (ROS) in the cerebral cortex and hippocampus of rat Aβ40 brain and the expression of nitric oxide synthase (iNOS) in hippocampus.

Figure 11 is a schematic diagram showing the deposition of Aβ40 after continuous infusion of Aβ40 to the hippocampus of rats;

Figures 12-1 to 12-3 show the preparation of the composition for treating and preventing Alzheimer's disease of the present invention.

Claims (2)

The invention relates to a method for preparing and preventing a composition of Alzheimer's disease, which comprises the following steps: (1) cleaning a rice (Oryza satiVa L., Japonica) and sterilizing in a high-pressure high-temperature environment, wherein The high-pressure high-temperature environment refers to a pressure environment of 121 ° C and 1 kg / cm ² ; (2) inoculate a specific red sputum strain into a culture medium and rotate at 30 ° C, 100 ~ 150 rpm (revolutions per minute ; rpm) and 48 hours after inoculation, wherein the inoculum culture medium is prepared by soaking at least 5 g of the dried rice in 100 ml of distilled sterile water; (3) stirring the inoculum at 30 ° C The medium is continuously provided with 20% distilled water for sterilization within 72 hours; (4) is suitably stirred within 96 hours with every 10 hours at 30 ° C to achieve ripening; (5) one of the collected fermentations The red sorghum fermented product is dried at 55 ° C to 60 ° C for 24 hours; (6) the dried red sorghum fermented product is ground into a powder and analyzed for compliance with a component ratio condition, wherein The component ratio condition means that the red mash fermentation product contained in a powder form contains a monacolins, an anti-inflammatory substance and an antioxidant substance, the optimal weight ratio of the three substances is 40:2:1; and (7) the prepared powder of the red peony ferment It is added to an edible beverage in a ratio of 1.0 to 4.0% to make a red sip drink with the treatment and prevention of Alzheimer's disease. The method for producing a composition for treating and preventing Alzheimer's disease according to claim 1, wherein the step of drying the dried red peony fermentate into a powder and performing analysis to meet a component ratio condition Further, the method further comprises the steps of filling the prepared powder of the red peony fermentate into a capsule or making a tablet.