KR20130070616A - A composition for prevention and treatment of dementia or parkinson's disease comprising extracts of morus alba or lycium chinense, or mixture thereof as an active ingredient - Google Patents

Abstract

PURPOSE: A composition containing a Morus alba, a Lycium chinense extract, or a mixture thereof for preventing and treating dementia or Parkinson's disease is provided to ensure cell protection effect, antioxidation effect, and anti-apoptotic effect. CONSTITUTION: A pharmaceutical composition for preventing and treating Parkinson's disease contains a Lycium chinense extract as an active ingredient. A pharmaceutical composition for preventing and treating Alzheimer's disease contains a mixture of a Morus alba extract and a Lycium chinense extract, as an active ingredient. A health functional food for preventing and treating Parkinson's disease contains the Lycium chinense extract as an active ingredient. A health functional food for preventing and treating Alzheimer's disease contains the mixture as an active ingredient. [Reference numerals] (AA) Morus alba extract(mg/Kg)

Description

A composition for prevention and treatment of dementia or Parkinson's disease comprising extracts of Morus alba or Lycium chinense, or mixture according as an active ingredient}

The present invention is heartache (scientific name: Morus alba ) or phalanx ( Lycium) The present invention relates to a pharmaceutical composition or health functional food for preventing and treating dementia or Parkinson's disease, which contains chinense extract or a mixture thereof as an active ingredient.

According to the National Statistical Office (NSO), the percentage of the elderly population in Korea is as follows: In 2000, the proportion of the population aged 65 and over accounted for 7.2% of total population, It is expected to enter the aging society. As the aging issue becomes a social issue, the public's interest in the characteristics of the elderly population, the welfare of the elderly such as housing, health, culture, and leisure has increased, and the demand for statistics is also increasing. The key to this change is that chronic degenerative diseases are becoming more of a problem than acute communicable diseases, which have been the cause of death for the past 50 years due to the increase in the aging population. Among chronic degenerative diseases, cerebrovascular death is a very important disease that is the second leading cause of mortality due to single disease.

Dementia is a disorder characterized by impairment of normal daily life in occupational, social, and interpersonal relationships, with memory impairment, dementia, disorientation, loss of comprehension, change in personality and emotions, . Alzheimer's disease, vascular dementia, and other dementias caused by alcoholism, trauma, and sequelae of Parkinson's disease, depending on the cause, is a pathological symptom that should be distinguished from normal aging. do. In addition, recent epidemiologic studies have reported that risk factors for cerebrovascular diseases such as hypertension, diabetes, hyperlipidemia, and heart disease increase the incidence of Alzheimer's as well as vascular dementia. It is true.

Alzheimer's disease (AD) is characterized by a loss of neurons and an extracellular senescent half-life with amyloid-beta (Aβ), a 39-43 amino acid peptide derived from amyloid precursor protein senile plaque. In vitro and in vivo studies have shown that A [beta] or A [beta] peptide fragments have toxic effects, suggesting that A [beta] plays an important role in the pathogenesis of AD (Butterfield et al . , Free Radical Biology and Medicine , 2002 , 32: 1050-1060; ButterfIeld et al ., Free Radical Biology and Medicine , 2007, 43: 658-677). At the time of culture, Aβ directly induces neuronal cell death and makes neuronal cells vulnerable to excitotoxic and oxidative damage. NMDA (N-methyl-D-aspartate receptor) receptors act as mediators of Aβ-binding selective substrate or Aβ-induced glutamate excitotoxicity. NMDA receptors are highly transmissive in particular ligand in Ca ^{2 +} - gate / overvoltage - is sensitive cation channels. Extensive increase in [Ca ^{2 +]} _i will lead directly to cell dysfunction, over-excitement or death. Thus, the neurotoxic effect of A? Is a noncompetitive NMDA receptor antagonist (5R.10S) - (+) - 5-methyl-10,11-dihydro-5H- dibenzo [a, d] cycloheptene- -Imine maleate (MK-801) [5R.10S) - (+) - 5-methyl-10,11-dihydro-5H-dibenzo (a, b) cyclohepten-5,10-imine maleate] as will be demonstrated by the report, Ca ^{+ 2} influx through NMDA receptors by exposure Aβ plays a crucial role in the neurotoxicity induced Aβ-. The formation of reactive oxygen species (ROS) is also believed to be involved in the onset of degenerative brain diseases. Some evidence supports the involvement of oxidative stress as an active factor in A [beta] -mediated neuropathy by triggering or facilitating neurodegeneration through a broad molecular shape that interferes with neuronal homeostasis. However, the clinical benefit of direct blockers of NMDA receptor antagonists and neuronal channels is controversial, as they lack the verifiable efficacy or have serious adverse effects.

Parkinson's disease is a chronic progressive degenerative disease of the nervous system characterized by stable tremor, stiffness, gait, and postural instability, which results in gradual loss of dopaminergic neurons in the subcortical nigra pars compacta (SNc) of the brain Exhibits neuropathologic features (Calne meat al ., 1983, Heikkila 1984). Parkinson's disease is estimated to be about 1% of the population at approximately 60 years of age or older.

Although the cause of Parkinson's disease is not clear, the 'multinomial hypothesis' that genetic and environmental factors interact with each other is the most commonly accepted. Most patients with Parkinson's disease develop without familial involvement, but about 10% are familial Parkinson's.

L-Dopa is currently commonly used as a symptomatic agent for increasing the reduced amount of dopamine in Parkinson's disease. After use, L-Dopa slows down Parkinson's disease and relieves clinical symptoms. However, L-Dopa has adverse effects such as involuntary exercise and vomiting after long-term use (Clarke and Deane, 2001). In addition, therapies used to treat Parkinson's disease include FDA-approved dopamine agonists, catechol-O-methyltransferase inhibitors (COMT inhibitors), and monoamine oxidase B ( monoamine oxidase B, MAO-B inhibitors) and anti-cholinergics.

Animal models used for the study of Parkinson's disease include 6-hydroxydopamine (6-OHDA), rotenon, 1-methyl-4-phenyl-1,2,3,6-tetrahydro There is an animal model using pyridine (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, MPTP). Among them, MPTP was first known to induce Parkinson's disease in humans by drug addicts in 1982, which is clinically similar to both humans as well as to humans in primates and mice, and is suitable for studying the pathophysiological changes of Parkinson's disease It has been evaluated as an experimental animal model. The mechanism of MPTP-induced dopaminergic neuronal injury has not yet been elucidated. Recently, however, MPTP exposure has been shown to increase inflammation in the cerebrum, and this inflammatory response has been reported to be an important process in pathological studies of Parkinson's disease . MPTP has been used as a useful Parkinson model since it causes acute degeneration of the nigrostriatal pathway in mice and monkeys. In addition, the animal model of MPTP is known to be suitable for studying the therapeutic effect of apoptosis due to mitochondrial dysfunction and oxidative stress as a stage of acute inflammation of Parkinson's disease or the neuroprotective effect of drugs have.

Heartache (scientific name: Morus alba , English name: white Mulberry) is a deciduous tree belonging to the Mulberry family (Moraceac), when the fruit of mulberry radish is magenta, it is harvested and used as a medicinal herb, and it is known to be used for the treatment of dizziness, tinnitus, browning, and beef Byung-Soo Kang et al., Herbology, Younglimsa, Seoul, Korea 2000), and Japan, it is known to apply to the treatment of tonic, pain medication, insomnia, tinnitus, dizziness, back pain, constipation, etc. (Namba, T., The Encyclopedia of Wakan-Yaku with Color Pictures Vol. 1 Hoikusha, Osaka, Japan, 1993). Dongbogam writes that it governs Sogal, benefits the five chapters, and gathers the mulberry tree's tablets. (Engbobogam Regional Committee, Dongbobogam, Namsan Party, Seoul, Korea, 2000). Heartache has been reported with various vitamins, organic acids, sugars, etc., and research reports on active ingredients are not known.

Jigolpi (scientific name: Lycium chinense (Chinese Matrimony vine) is a root bark of goji berry tree and contains 0.08% of betaine and 1.07% of saponin, which is known to be effective for antipyretic, diabetes and lowering blood pressure. It has been shown to show a strong inhibitory effect against the typhoid group, paratyphoid A bacteria, and Prexena bacteria. The phalanges of these phalanxes are produced as a by-product when renewing the species of Goji, but only a small amount is used as a medicinal substance, and most of them are discarded, and studies on cerebrovascular cerebrovascular diseases are not known. .

Therefore, the present inventors are trying to develop a natural substance having a therapeutic and prophylactic effect on dementia or Parkinson's disease, and the extract of heartache or phalanx is mixed with beta amyloid (Amyloid-β) in the cerebral cortex and hippocampal. Cell protection against Abeta, protection against Thapsigargin, toxic to Endoplasmic reticulum stress, mitochondrial membrane potential, reactive oxygen species (ROS), bcl-2, bax and cas The anti-apoptosis effect and the protective effect on neurons through MAP-2 positive cell count were confirmed through the measurement of Caspase-3, and 6-OHDA (6-hydroxydopamine) induced toxicity. The present invention was completed by revealing the antioxidant and anti-apoptotic effects on the present invention, which can be usefully used for the prevention and treatment of dementia or Parkinson's disease.

An object of the present invention is upset (scientific name: Morus alba ) or phalanx ( Lycium) It is to provide a composition for the prevention and treatment of dementia or Parkinson's disease, and a dietary supplement for the prevention and improvement of dementia or Parkinson's disease containing chinense extract, or a mixture thereof as an active ingredient.

In order to achieve the above object, the present invention is the heart (morus name: Morus alba ) provides a composition for preventing and treating dementia containing the extract as an active ingredient.

The invention also jigolpi (scientific name: Lycium It provides a composition for preventing and treating dementia or Parkinson's disease containing chinense ) extract as an active ingredient.

The present invention also provides a composition for the prevention and treatment of dementia, containing a mixed extract of the heart and the phalanges as an active ingredient.

The present invention also provides a health functional food for preventing and improving dementia containing the extract of the heart as an active ingredient.

In addition, the present invention provides a functional food for preventing and improving dementia or Parkinson's disease containing the extract of the phalanges as an active ingredient.

In addition, the present invention provides a dietary supplement for preventing and improving dementia, which contains a mixed extract of lettuce and phalanges as an active ingredient.

Upset of the present invention (the scientific name: Morus alba ) or phalanx ( Lycium) chinense ) extract, or mixtures thereof, confirmed the cellular protective effect against amyloid-β (Abeta) -induced toxicity and the cytoprotective effect against Thapsigargin, and also 6-OHDA (6-hydroxydopamine) -induced toxicity. By confirming the antioxidant effect and anti-self killing effect on the heart, or extracts of the phalanges, or mixtures thereof, the pharmaceutical composition for the prevention and treatment of dementia or Parkinson's disease or the development of a health food for the purpose Can be used effectively.

1 is a heartache (scientific name: Morus alba ) is a graph showing the results of the passive avoidance task after the extract was treated.
Figure 2 is a diagram showing the change in neuroprogenitor cells (doublecortin; DCX) after treating the heart extract.
Figure 3 is a diagram showing the change in the growth of neurites (neurite) after processing the heart extract.
Figure 4 is a graph measuring the level of NGF (nerve growth factor; nerve growth factor) after treatment of the heart extract.
5 is a graph showing the cell viability after treatment of the heart extract.
Figure 6 is a graph showing the cell viability of the heart beat extract pretreatment for beta amyloid (Abeta) toxic treatment.
Figure 7 is a graph showing the anti-self killing effect of the lettuce extract.
8 is a graph showing the number of MAP-2 positive cells.
Figure 9 is a graph showing cell viability using tapsigargin for the heart extract.
10 is jigolpi (scientific name: Lycium chinense ) is a graph showing cell viability in cortical cells of the extract.
Figure 11 is a graph showing the cell viability in hippocampus (hippocampus) cells of the phalanx extract.
Figure 12 is a graph showing the cell survival rate using top sigajin for the phalanges extract.
Figure 13 is a graph showing the cell survival rate of the phalanx golgol extract for 6-OHDA (6-hydroxydopamine) induced toxicity.
14 is a graph showing the antioxidant effect of the phalanges extracts on 6-OHDA induced toxicity.
Figure 15 is a graph showing the anti-self killing effect of the phalanges extracts on 6-OHDA induced toxicity.
Figure 16 is a diagram showing the protective effect of the phalanx of the phalanx extracts against 6-OHDA or MPP + toxicity.
Figure 17 is a graph showing the behavioral test mitigation effect of the phalanges extract 6-OHDA toxicity.
18 is a graph showing the cytoprotective effect of the mixture of the heart and phalanges.

Hereinafter, the present invention will be described in detail.

The present invention is heartache (scientific name: Morus alba ) provides a composition for preventing and treating dementia containing the extract as an active ingredient.

The heart can be used without limitation, such as cultivated or commercially available.

The dementia is preferably Alzheimer's disease, but is not limited thereto.

The lettuce extract is preferably prepared in the following steps, but is not limited thereto.

1) extracting the dried heart by adding an extraction solvent;

2) filtering the extract of step 1); And

3) Concentrating the filtered extract of step 2) under reduced pressure.

In the above method, the extraction solvent of step 1) is preferably a solvent selected from water, an alcohol or a mixture thereof, preferably a C ₁ to C ₂ lower alcohol or a mixed solvent thereof, and a 70% aqueous ethanol solution But it is not limited thereto. The amount of the extraction solvent is preferably 5 to 15 times the dry weight of the heart, and more preferably 7 to 10 times, but not always limited thereto. The extraction method may be an extraction method such as hot water extraction, immersion extraction, reflux cooling extraction, or ultrasonic extraction, but is not limited thereto. The extraction temperature is preferably in the range of 10 ° C to 100 ° C, more preferably room temperature. The extraction time is preferably 30 minutes to 3 hours, more preferably 1 to 2 hours, but is not limited thereto. The number of times of extraction is preferably 1 to 5 times, more preferably 3 times, but is not limited thereto.

In the above method, the reduced pressure concentration in step 3) is preferably a vacuum rotary evaporator, but is not limited thereto. The drying is preferably performed by lyophilization, but is not limited thereto.

The present inventors conducted a passive avoidance test to confirm the simple cognitive function of the heart extract extract, it was confirmed that showing the significant memory enhancement effect when administering the heart extract (see Fig. 1).

In addition, in order to confirm the simple cognitive function of the extract of the heart, extracts were measured by immunohistochemical staining of the changes in the neural precursor cells (doublecortin) and neurites (neurite). It was confirmed that it showed a significant increase in the projections (see FIGS. 2 and 3).

In addition, in order to confirm the effect of improving the simple cognitive function of the lettuce extract, as a result of measuring the NGF (nerve growth factor; nerve growth factor) level, it was confirmed that the NGF significantly increased when administering the lettuce extract (see Fig. 4). ).

Therefore, the extract of the heart of the present invention shows a significant memory enhancing effect, passive increase in neuronal progenitor cells and neurites, and a significant increase in NGF as a result of passive avoidance test, preventing and improving diseases related to cognitive dysfunction Or as an active ingredient of a therapeutic composition.

In addition, in order to confirm the protective effect in the cells of the lettuce extract, the cell viability was measured according to the beta amyloid (Abeta) -induced toxicity, the administration of the lettuce extract significantly increased the cell viability compared to the control group It was confirmed that (see Fig. 5 and 6).

In addition, the present inventors measured the cell viability using Thapsigargin, which is toxic to ER stress (Endoplasmic reticulum stress), in order to confirm the protective effect in the cells of the heart beat extract, the cell viability reduced by Topsijin It was confirmed that the dama extract extract was protected by simultaneous treatment (see FIG. 9).

In addition, the present inventors measured the mitochondrial membrane potential, reactive oxygen species (ROS), bcl-2 and bax in order to confirm the anti-apoptosis effect of the lettuce extract, the lettuce extract In the pretreatment group, the inhibition of mitochondrial membrane potential disruption affecting apoptosis signaling, the reduction of ROS that promotes apoptosis, the reduction of bax, a protein that induces apoptosis, and the increase of the anti-apoptosis protein, bcl-2, were confirmed. (See FIG. 7).

In addition, the present inventors measured the number of MAP-2 positive cells in order to confirm the protective effect on the neurons of the heart extract extract, it was confirmed that the MAP-2 positive cell number reduced by Abeta is protected by the heart extract extract pretreatment ( 8).

Therefore, the lettuce extract of the present invention protects the cells against the toxicity of beta amyloid and top sigajin, has an anti-self killing effect, and has a protective effect on neurons, so as to be useful as an active ingredient of a composition for preventing and treating dementia. Can be used.

The invention also jigolpi (scientific name: Lycium It provides a composition for preventing and treating dementia or Parkinson's disease containing chinense ) extract as an active ingredient.

The phalanges can be used without limitation, such as those grown or commercially available.

The dementia is preferably Alzheimer's but is not limited thereto.

The phalanges extract is preferably prepared in the following steps, but is not limited thereto.

1) extracting the dried phalanges by adding an extraction solvent;

2) filtering the extract of step 1); And

3) Concentrating the filtered extract of step 2) under reduced pressure.

In the above method, the extraction solvent of step 1) is preferably a solvent selected from water, an alcohol or a mixture thereof, preferably a C ₁ to C ₂ lower alcohol or a mixed solvent thereof, and a 70% aqueous ethanol solution But it is not limited thereto. The amount of the extraction solvent is preferably 5 to 15 times the dry weight of the phalanges, and more preferably 7 to 10 times, but not always limited thereto. The extraction method may be an extraction method such as hot water extraction, immersion extraction, reflux cooling extraction, or ultrasonic extraction, but is not limited thereto. The extraction temperature is preferably in the range of 10 ° C to 100 ° C, more preferably room temperature. The extraction time is preferably 30 minutes to 3 hours, more preferably 1 to 2 hours, but is not limited thereto. The number of times of extraction is preferably 1 to 5 times, more preferably 3 times, but is not limited thereto.

In the above method, the reduced pressure concentration in step 3) is preferably a vacuum rotary evaporator, but is not limited thereto. The drying is preferably performed by lyophilization, but is not limited thereto.

The present inventors measured the cell viability according to beta amyloid-induced toxicity in order to confirm the protective effect in the cells of the phalanx extract, it was confirmed that administration of the phalanx extract significantly increased the cell viability compared to the control (Fig. 10 and FIG. 11).

In addition, the present inventors measured the cell viability using the ER stress toxic Thapsigin, in order to confirm the protective effect in the cells of the phalanx extract, the cell viability reduced by the top sigajin is protected by simultaneous treatment of the phalanx extract It confirmed that it became (refer FIG. 12).

In addition, the present inventors measured the cell survival rate using 6-OHDA (6-hydroxydopamine), which causes dopamine neurological lesions in the injected site, in order to confirm the protective effect in the cells of the phalanx extract, to protect the cells and Survival was confirmed to increase significantly compared to the control (see Figure 13).

In addition, the present inventors measured the ROS and NO increased by 6-OHDA in order to confirm the antioxidant effect of the phalanges of the extract, it was confirmed that the extract of the phalanges inhibits the overproduction of ROS and NO, and inhibits the activity of GSH. (See FIG. 14).

In addition, the present inventors confirmed the mitochondrial membrane potential, cytochrome C concentration and caspase-3 activity in order to confirm the anti-self killing effect of the phalanges extract, caspase in the pretreatment of the phalanges extract It was confirmed that inhibiting the -3 activity, inhibited the disturbance of the membrane potential of the meat cone and suppressed the increase of cytochrome C concentration (see Fig. 15).

In addition, the present inventors measured the number of TH-positive neurons in order to confirm the protective effect of the phalanx extract on dopaminergic cells, the number of TH-positive neurons in the pretreatment of the phalanx extract Was found to increase significantly (see FIG. 16).

In addition, the present inventors measured the rotation test in order to confirm the behavioral test mitigation effect on 6-OHDA toxicity of the phalanx extract, the phalanx extract was confirmed that the behavioral symptoms were alleviated compared to the toxic control group (See FIG. 17).

Therefore, the phalanges extract of the present invention protects the cells against beta amyloid, top cigar, 6-OHDA and MPP + toxicity, has antioxidant and anti-apoptotic effects, and also has behavioral alleviating effects on 6-OHDA toxicity. Therefore, it can be usefully used as an active ingredient of a composition for preventing and treating dementia or Parkinson's disease.

The present invention also provides a composition for the prevention and treatment of dementia, containing a mixed extract of the heart and the phalanges as an active ingredient.

The mixture is a mixture of the heart beat and the phalanges extract preferably in a 1: 1 ratio, but is not limited thereto.

The mixed extract may be used by mixing each extract, or both prepared by extracting the heart and the phalanges together.

The heartache and phalanges can be used without limitation, such as those grown or commercially available.

The dementia is preferably Alzheimer's but is not limited thereto.

As the solvent of the extract, it is preferable to use a solvent selected from water, an alcohol or a mixture thereof, preferably a lower alcohol of C ₁ to C ₂ or a mixed solvent thereof, and more preferably, an aqueous 70% ethanol solution is used. It is not limited to this. The amount of the extraction solvent is preferably 5 to 15 times the weight of dry heart or phalanges, and more preferably 7 to 10 times, but not always limited thereto. The extraction method may be an extraction method such as hot water extraction, immersion extraction, reflux cooling extraction, or ultrasonic extraction, but is not limited thereto. The extraction temperature is preferably in the range of 10 ° C to 100 ° C, more preferably room temperature. The extraction time is preferably 30 minutes to 3 hours, more preferably 1 to 2 hours, but is not limited thereto. The number of times of extraction is preferably 1 to 5 times, more preferably 3 times, but is not limited thereto.

In the above method, the reduced pressure concentration in step 3) is preferably a vacuum rotary evaporator, but is not limited thereto. The drying is preferably performed by lyophilization, but is not limited thereto.

The present inventors measured the cell viability according to the beta amyloid-induced toxicity in order to confirm the protective effect in the cells of the mixture of the heart and the phalanges, the cell survival rate was significantly higher than the control group when pre-treated with the mixture of the heart and the phalanges It was confirmed that the increase by (see Figure 18).

Therefore, the mixture of the heartache and phalanges of the present invention has an effect of protecting the cells against beta amyloid toxicity, so it can be usefully used as an active ingredient of a composition for preventing and treating dementia.

The composition of the present invention preferably comprises 0.1 to 90% by weight of the lettuce extract or phalanges extract, or a mixture thereof, based on the total weight of the composition, but is not limited thereto.

The composition of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the preparation of pharmaceutical compositions.

The composition of the present invention may be administered orally or parenterally, and when parenteral administration, it is preferable to select external or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection injection method. It is not limited thereto.

The composition of the present invention can be used in the form of oral formulations, external preparations, suppositories, and sterile injectable solutions, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, respectively, according to conventional methods. have. Carriers, excipients and diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, Methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations can be used in at least one excipient such as starch, calcium carbonate (calcium carbonate) carbonate, sucrose or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The preferred dosage of the composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the composition is preferably administered at a dose of 0.0001 to 1 g / kg per day, preferably 0.001 to 200 mg / kg per day, but is not limited thereto. The administration may be administered once a day, or may be divided several times. The dose is not intended to limit the scope of the invention in any way.

In another aspect, the present invention provides a health food for preventing and improving dementia containing the extract of the heart as an active ingredient.

The dementia is preferably Alzheimer's but is not limited thereto.

The extract of the present invention protects the cells against the toxicity of beta amyloid and top siginine, has an anti-self killing effect, and has a protective effect on neurons, so it is usefully used as an active ingredient of health food for preventing and improving dementia. Can be.

In addition, the present invention provides a health food for preventing and improving dementia or Parkinson's disease containing the phalanges extract as an active ingredient.

The dementia is preferably Alzheimer's but is not limited thereto.

The phalanx extract of the present invention protects the cells against beta amyloid, top cigar, 6-OHDA and MPP + toxicity, has antioxidative and anti-apoptotic effects, and also has behavioral alleviating effects on 6-OHDA toxicity. Or it can be usefully used as an active ingredient in health food for preventing and improving Parkinson's disease.

In another aspect, the present invention provides a health food for preventing and improving dementia containing a mixed extract of lettuce and phalanges as an active ingredient.

The dementia is preferably Alzheimer's but is not limited thereto.

The mixed extract of the heart beat and the phalanges of the present invention has an effect of protecting the cells against beta amyloid toxicity, and thus can be usefully used as an active ingredient for health foods for preventing and improving dementia.

There is no particular limitation on the kind of the food. Examples of the food include dairy products including drinks, meat, sausage, bread, biscuits, rice cakes, chocolates, candies, snacks, confectionery, pizza, ramen noodles, other noodles, gums, ice cream, various soups, And a combination thereof, all of which include health foods in a conventional sense.

The heartache or phalanges extracts of the present invention, or mixtures thereof, may be added as is to food or used together with other food or food ingredients, and may be appropriately used according to conventional methods. The amount of the active ingredient to be mixed can be suitably determined according to its use purpose (for prevention or improvement). Generally, the amount of the extract in the health food may be 0.01 to 15% by weight of the total food, and the health beverage composition may be added in a proportion of 0.02 to 5 g, preferably 0.3 to 1 g, based on 100 ml have. However, in the case of long-term intake intended for health and hygiene purposes or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

The health functional beverage composition of the present invention is not particularly limited to other ingredients other than the above containing the heart beat or phalanges extract, or mixtures thereof as essential ingredients in the indicated ratios, and adds various flavors or natural carbohydrates, such as ordinary drinks. It may contain as a component. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. .

In addition to the above-mentioned foods, the food of the present invention may contain flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and heavies (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the extract of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the extract of the present invention.

Hereinafter, the present invention will be described in detail with reference to Examples, Experimental Examples and Preparation Examples.

However, the following Examples, Experimental Examples and Preparation Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Examples, Experimental Examples, and Preparation Examples.

< Example  1> Heartache Morus alba ) And Phalanges (Scientific name: Lycium chinense ) Preparation of extract

<1-1> Preparation of Lettuce Extract

100 mL of 70% ethanol was added to 100 g of the heartbeat purchased from Jeong-Do (Seoul), and leached under stirring for 24 hours, followed by filtering using Whatman filter paper # 2. . The filtrate was concentrated under reduced pressure at 50 ° C. (Rotavapor R-200, heating bath B-490, BUCHI; Flawil, Switzerland). The sample was freeze-dried and stored at -20 ° C., and was used in the preparation of experiments. The yield was 20.53. Was%.

<1-2> Phalanges  Preparation of extract

100 mL of 70% ethanol was added to 100 g of the heartbeat purchased from Jeong-Do (Seoul), and leached under stirring for 24 hours, followed by filtering using Whatman filter paper # 2. . The filtrate was concentrated under reduced pressure at 50 ° C. (Rotavapor R-200, heating bath B-490, BUCHI; Flawil, Switzerland). The sample was freeze-dried and stored at −20 ° C., and used as a preparation in the experiment. 19.46%.

<1-3> heartache and Phalanges  Preparation of a Mixture of Extracts

It was prepared by mixing the lettuce extract and the phalanges extract prepared in Examples <1-1> and <1-2> in a ratio of 1: 1. In addition, all were stored at -20 ° C experimentally used for preparation.

< Experimental Example  1> Confirm the effect of improving the simplicity of heartache

<1-1> Experimental Animal Preparation

Experimental animals used male ICR mice (mice) (25 ~ 30 g, Orient). The wound was used after suspension in 10% dimethyl sulfoxide (DMSO). The drugs were all used on the day of the experiment, and divided into three groups as shown in Table 1. Each group was orally administered with solvents and samples for 7 days. After 1 hour of oral administration on the 6th and 7th days, the training trial and the acquisition trial were performed in a manual avoidance test. On the 8th day, a retention trial was conducted.

group p.o. Control group 10% DMSO ME-100 mg / kg 10% DMSO with 100 mg / kg of heart ME-500 mg / kg 10% DMSO with 500 mg / kg of heart

<1-2> Manual avoidance experiment Passive avoidance task )

Training trials and tests were conducted in two separate and independent bright and dark square boxes. Bright areas (20 × 20 × 20 cm) were illuminated with 50 W incandescent bulbs. The light and dark areas (20 × 20 × 20 cm) were spaced 1 cm apart with 2 mm stainless steel rods and laid out the length of the area. The zone was separated by a guillotine door (5 × 5 cm). The mice are placed in the first bright zone and the door between them opens after 10 seconds. When the mice entered the dark area, the door was automatically closed and immediately subjected to an electric foot shock (0.3 mA) through a stainless steel rod for 3 seconds (Acquisition trial). 24 hours after the accumulation trial, the mice were placed in bright areas to measure the retention trial. The time was measured for both the acquisition trial and the retention trial until the rats had all four feet from the light room to the dark room. The mice were orally administered with solvent and heart beat extract 1 hour before the accumulation trial.

As a result, as shown in Figure 1, it was confirmed that statistically significant memory enhancing (memory enhancing) effect when oral administration of the heart extract extract at 100 and 500 mg / kg for 7 days (Fig. 1).

<1-3> Immunohistochemical Staining

Experimental Example <1-1> For immunohistochemical staining, the animals were perfused with 1X PBS (phosphate buffer saline), fixed with 4% paraformaldehyde, and brains were extracted. It was fixed and put in a 30% squarose solution (sucrose) solution was stored every two days until the frozen section at 4 ℃. Thereafter, brain tissue was frozen in a cryostat by dropping the OCT (optimal cutting temperature) compound at -20 ° C, and then made into sections having a thickness of 30 µm and stored in 4 ° C. Immunohistostaining was performed with the hippocampus, and the tissues washed with PBS were treated with 1% H ₂ O ₂ for 15 minutes to prevent nonspecific reactions. 0.05M PBS, 1.5% normal goat serum , 0.5 mg / ml bovine serum albumin, 0.3% triton X-100 and a primary antibody (goat anti-DCX primary antibody, 1: 500) were reacted at 4 ° C. for 24 hours. After removing the primary antibody, the tissue was reacted with a peroxidase-linked secondary antibody (1: 200) for 90 minutes, and then diluted with ABC in a buffer solution for about 1 hour at room temperature. After washing three times with PBS, and then developed with 0.02% DAB and 0.0 1% H ₂ O ₂ slide samples were made through ethanol and xylene dehydration process.

As a result, as shown in Figs. 2 and 3, neuronal progenitor cells (doublecortin; DCX) were significantly increased and oral neurite was significantly increased after oral administration at 100 and 500 mg / kg for 7 days. Confirmed. Therefore, it was confirmed that the long-term administration of heartache is effective in improving the simple cognitive function (Figs. 2 and 3).

<1-4> NGF ( nerve growth factor ; Neural growth factor) level measurement

NGF levels were measured to determine the effect of cognitive improvement on heartache. In order to measure NGF levels, the brains of the experimental animals of Experimental Example <1-1> were extracted, and the hippocampus was homogenated. Chemikine ^TM purchased from Chemicon International (Temecula, CA, USA) NGF levels were measured using an NGF sandwich enzyme-linked immunosorbentassay (ELISA) kit.

As a result, as shown in FIG. 4, it was confirmed that NGF significantly increased upon oral administration at 100 and 500 mg / kg of heartbeat for 7 days. Therefore, it was confirmed that the long-term administration of heart heart is effective in improving the NGF level and thus simple cognitive function (Fig. 4).

< Experimental Example  2> In vitro ( in vitro A) of heartache extract Anti-Alzheimer's  Check the effect

<2-1> cerebral cortex ( Cortical ) On cell protection effect

Amyloid plaque, produced by the accumulation of beta amyloid-β (Abeta), is known as a substance that accumulates in the brain and kills nerve cells, and is used for Alzheimer's research. Abeta (25-35) artificially aggregated (aggregated) material to produce a toxicity to the cerebral cortical cells by using the MTT assay (assay) to determine the cytoprotective effect of the heart extract. Specifically, first, only the cerebral cortex is isolated from the 18-day-old fetus of Sprague-Dawley rats (Orient Bio, Seoul), and then mechanically decomposed to obtain cells, and then poly-L- Cells were seeded at 1 × 10 ⁴ / well in 96 well plates pre-coated with lysine (Poly-L-lysine) and cultured for 7 days. Subsequently, the beta amyloid (Abeta, 8 μg) was treated with 0.1% 1, 10 or 100 μg / mL of the persimmon extract in 0.1% DMSO / B27 free neuronal media. Or non-treated and incubated for 24 hours, after 3 hours of 1 mg / mL MTT treatment, formazan was dissolved in DMSO and absorbance was measured at 570 nm.

As a result, as shown in Figures 5 and 6, when treated with a concentration of 0.1, 1, 10 or 100 ㎍ / mL in the cerebral cortex cells alone, it was confirmed that does not affect the cell viability. In addition, in the case of Abeta toxicity treatment, the cell viability was reduced to 73.68 ± 1.11% compared to the control group, and in the case of the lettuce extract pretreatment group showed a cytoprotective effect at a concentration of 10 or 100 ㎍ / mL (Figs. 5 and 6). .

<2-2> hippocampus ( hippocampal ) On cell protection effect

In order to confirm the cytoprotective effect of the heart extract, only the hippocampus was isolated from the 18-day-old fetus of Sprague Dawley rats (Orient Bio, Seoul), and mechanically decomposed to obtain cells, followed by poly-L-lysine. Cells were seeded at 1 × 10 ⁴ / well in 96 well flights precoated with 7 days. Subsequently, after 30 minutes of treatment with 0.1, 1, 10 or 100 μg / mL of the persimmon extract in 0.1% DMSO / B27 free neuronal basal medium, beta amyloid (Abeta, 8 μg) was treated or non-treated in total 24 After incubation for 3 hours, 3 mg of 1 mg / mL MTT treatment, formazan was dissolved in DMSO, and absorbance was measured at 570 nm.

As a result, it was confirmed that the cell viability was increased at 1, 10 or 100 ㎍ / mL concentration when compared to the concentration of 0.1, 1, 10 or 100 ㎍ / mL in the seaweed extract alone in the hippocampal cells. In addition, Abeta inhibited the survival of 52.97 ± 0.97% hippocampal cells as compared to the control group, and showed a statistically significant protective effect in pretreatment concentrations of 0.1, 1, 10 or 100 ㎍ / mL of the heart extract.

<2-3> anti-self killing of the lettuce extract ( anti - apoptosis ) effect

In order to confirm the anti-self killing effect of the extract of the heart, extracts were inoculated in a 60 mm plate coated with PLL-coated hippocampal cells, and cultured for 7 days, and the extract of 10 μg / mL in a 0.1% DMSO / B27 free neuronal base medium was used. After 30 minutes of treatment, Abeta (8 ㎍) treated with a total 18 hours of incubation, mitochondrial membrane potential (mitochondrial by measuring the fluorescence of the red and green wavelengths using mitochondrial-specific dye JC-1 membrane potential) was observed.

As a result, it was confirmed that the mitochondrial membrane potential, which affects apoptosis signal transduction, was maintained by the heart extract extract as shown in FIG. 7 (FIG. 7).

<2-4> Anti-Apoptosis of Lettuce Extracts anti - apoptosis ) effect

In order to confirm the anti-self killing effect of the extract of the heart, extracts were inoculated in a 60 mm plate coated with PLL-coated hippocampal cells, and cultured for 7 days, and the extract of 10 μg / mL in a 0.1% DMSO / B27 free neuronal base medium was used. After 30 minutes of treatment, Abeta (8 ㎍) treated with a total of 18 hours incubation, cells were collected and divided into mitochondrial and mitochondrial (cytosol), the protein was extracted and the change was observed using the antibody.

As a result, the mitochondrial membrane potential, which affects apoptosis signaling, is maintained by the heartworm extract pretreatment, as shown in FIG. 7, whereby the concentration of cytochrome C in the cytosol was compared with the Abeta group. It was confirmed that low in (Fig. 7).

<2-5> Anti-Apoptosis of Lettuce Extracts anti - apoptosis ) effect

In order to confirm the anti-self killing effect of the heart extract, the amount of reactive oxygen species (ROS), bcl-2 and bax produced after drug and toxic treatment of hippocampal cells with DCF-DA fluorescent dye (dye) Was measured.

As a result, as shown in Figure 7 when treated with Abeta compared to the control, an increase in ROS that promotes apoptosis, a decrease in the anti-apoptosis protein (bcl-2) and cells While it was confirmed that bax, which is a pro-atpotosis protein, was increased, pretreatment with the heart beat extract showed a decrease in ROS production, an increase in bcl-2, and a decrease in bax (FIG. 7).

<2-6> Anti-self-killing of Lettuce Extracts anti - apoptosis ) effect

In order to confirm the anti-apoptotic effect of the extracts, the inoculated cells were inoculated into a 60 mm plate coated with PLL-coated hippocampal cells, and cultured for 7 days, and 10 µg / mL of the extracts were added to a 0.1% DMSO / B27 free neuronal base medium. After 30 minutes of treatment, the cells were incubated with Abeta (8 ㎍) for 24 hours, and then the cells were collected to collect caspase-3 activity and cleavaged caspase-3 form. ) Were identified using kit (Bio Vision, USA) and Western blot, respectively.

As a result, caspase-3 activity and cleavaged caspase-3 form, which are involved in apoptosis, were reduced by pretreatment of the wick extract as shown in FIG. 7. By confirming that, the anti-self killing effect of the lettuce extract (Fig. 7) was confirmed.

<2-7> MAP -2 Positive cell count  Confirm

In order to measure the protective effect on neurons using MAP-2 antibody, a neuronal marker, hippocampal cells were inoculated in 24 wells coated with PLL and cultured for 7 days, and 0.1% DMSO / B27 free. After treatment for 10 minutes of 10 ㎍ / mL heart extract extract to the neuronal base medium, Abeta (8 ㎍) was incubated for 24 hours. After the reaction was fixed and stained with MAP-2 antibody (antibody) and the number of positive cells were counted and compared with the control.

As a result, as shown in FIG. 8, it was confirmed that the number of MAP-2 positive cells significantly reduced by Abeta was protected by the heart extract extract pretreatment (FIG. 8).

<2-8> Top City ( Thapsigargin ) For

In order to confirm cell viability using TopSigazine, which is toxic to ER stress (Endoplasmic reticulum stress), inoculated with hippocampal cells into PLL-coated 96 well flight at 1 × 10 ⁴ / well for 7 days, followed by 0.1% DMSO / B27-free neuronal basal medium was treated with 0.1, 1, 10, or 100 μg / mL of the persimmon extract, followed by incubation for 24 hours with or without treatment with Topsigin (25 nM). After 3 hours of 1 mg / mL MTT treatment, formazan was dissolved in DMSO and absorbance was measured at 570 nm.

As a result, as shown in Figure 9, it was confirmed that the cell viability reduced by the top sigajin is protected by the co-treatment extract (Fig. 9).

< Experimental Example  3> in vitro Phalanges  Extract Anti-Alzheimer's  Check the effect

<3-1> cerebral cortex ( Cortical ) On cell protection effect

In order to confirm the cytoprotective effect of the phalanges of the golgolpi extract, only cerebral cortical sites were isolated from 18-day-old fetus of Sprague-Dawley rats (Orient Bio, Seoul) and mechanically decomposed. Thereafter, the cells were seeded at 1 × 10 ⁴ / well in 96 well flights previously coated with poly-L-lysine, and then cultured for 7 days. Subsequently, after 30 minutes of treating the phalanx with 0.1, 1, or 10 μg / mL in 0.1% DMSO / B27 free neuronal base medium, incubated for 24 hours with beta amyloid (Abeta, 8 μg) or non-treated. After 3 hours of 1 mg / mL MTT treatment, formazan was dissolved in DMSO and absorbance was measured at 570 nm.

As a result, as shown in Figure 10, when treated with a concentration of 0.1, 1 or 10 ㎍ / mL with the phalanx extract alone in cerebral cortical cells, it was confirmed that the cells increase at a concentration of 1 or 10 ㎍ / mL, Abeta toxicity In the case of treatment, the cell viability was decreased compared to the control group, and in the case of the phalanx extract pretreatment group, it was confirmed that there was a cell protective effect at a concentration of 1 or 10 μg / mL (FIG. 10).

<3-2> hippocampus ( hippocampal ) On cell protection effect

In order to confirm the cytoprotective effect of the phalanx extract, only the hippocampus was isolated from the 18-day-old fetus of Sprague Dawley rats (Orient Bio, Seoul), and mechanically decomposed to obtain cells, followed by poly-L-lysine. Cells were seeded at 1 × 10 ⁴ / well in 96 well flights precoated with 7 days. Subsequently, after 30 minutes of treatment with 0.1, 1 or 10 μg / mL of the persimmon extract in 0.1% DMSO / B27 free neuronal basal medium, incubated for 24 hours with beta amyloid (Abeta, 8 μg) or non-treated. After 3 hours of 1 mg / mL MTT treatment, formazan was dissolved in DMSO and absorbance was measured at 570 nm.

As a result, as shown in Figure 11, when treated with a concentration of 0.1, 1 or 10 ㎍ / mL in the hippocampus extract alone in the hippocampal cells it was confirmed that the cell viability increased at a concentration of 1 or 10 ㎍ / mL compared to the control. In addition, Abeta inhibited the survival of hippocampal cells compared to the control group, and confirmed a statistically significant protective effect in the concentration pre-treatment of phalanges extract 0.1, 1 or 10 ㎍ / mL (Fig. 11).

<3-3> On top  For effect

In order to confirm cell viability using ER stress toxic thapciazine, inoculated with hippocampal PLL-coated 96 well flight at 1 × 10 ⁴ / well for 7 days, and then cultured with 0.1% DMSO / B27-free neuronal base. After treating the phalanx extract to the medium at 0.01, 0.1, 1, or 10 μg / mL, and incubated for 24 hours by treatment or non-treatment with Topsigin (25 nM). After 3 hours of 1 mg / mL MTT treatment, formazan was dissolved in DMSO and absorbance was measured at 570 nm.

As a result, as shown in Figure 12, it was confirmed that the cell viability reduced by the top sigajin is protected at 0.01 to 10 ㎍ / mL concentration by simultaneous treatment of the phalanges extract (Fig. 12).

< Experimental Example  4> in vitro Phalanges  Extract Anti-Parkinson's disease  Check the effect

<4-1> Phalanges  Cell Protection Effect of Extracts

To determine the cell viability of the phalanx extracts against 6-OHDA (6-hydroxydopamine) -induced toxicity causing dopamine neurological lesions at the site of injection, dopamine-releasing fibroblastoma SH-SY5Y cells in 96-well flight at 2x10 ⁴ / well After inoculation, two days were incubated, and each drug dissolved in 0.1% DMSO / FBS free medium was pretreated with 0.01, 1, or 10 μg / mL for 18 hours, followed by 6-OHDA treatment for 6 hours. After completion of the reaction, the cells were treated with MTT 1 mg / mL for 3 hours, and then absorbed at 570 nm using a spectrophotometer to confirm cell viability.

As a result, as shown in Figure 13, 24 hours treatment with the phalanx extract alone in SH-SY5Y cells did not affect cell viability, 18 hours pre-treatment of the phalanx extract was 0.01-10 ㎍ for 6-OHDA toxicity A significant protective effect was confirmed at the concentration of / mL (Fig. 13).

<4-2> Phalanges  Antioxidant Effect of Extracts

After treatment with SH-SY5Y cells with 0.1-10 μg / mL of drug and 6-OHDA, the culture medium was recovered, and the culture medium was reacted with the same amount of grease reagent, and the absorbance was measured at 540 nm with a spectrophotometer. NO concentration was determined by drawing a standard curve using sodium nitrite, and 0.1-μg / mL of drug and 6-OHDA were treated in SH-SY5Y cells to confirm ROS change, and then 20 μM H₂DCF. After adding -DA and incubation for 30 minutes, washed with PBS, measured by fluorescence excitation 495 nm, emission 530 nm. In addition, after treatment of SH-SY5Y cells with 0.1-1 μg / mL of drug and 6-OHDA, cells were collected and then treated with non-enzymatic antioxidant defense system. GSH levels that play a role were measured using the GSH Essay Kit (Dojindo).

As a result, as shown in Figure 14, the phalanx extract showed a significant protective effect while suppressing the overproduction of ROS and NO increased by 6-OHDA at a concentration of 0.1-10 / mL. In addition, the effect of inhibiting the activity of GSH by 6-OHDA at concentrations of 0.1 and 1 ㎍ / mL was confirmed (Fig. 14).

<4-3> Phalanges  Confirmation of anti-self killing effect of extract

To observe the change in mitochondrial membrane potential, SH-SY5Y cells were treated with 1-10 μg / mL of drug and 6-OHDA, and then the reaction cells were collected and mitochondrial-specific stained JC-1 was used. Fluorescence of the red and green wavelengths was measured, and 0.1-μg / mL drug and 6-OHDA were treated in SH-SY5Y cells to measure caspase-3 activity. Caspase-3 activity was measured using the Caspase-3 kit. In addition, after treatment with 0.1-1 μg / mL of drug and 6-OHDA to SH-SY5Y cells, the cells were collected and the cytochrome C release was measured by cytosolic / mitochondrial fraction kit (cytosolic). Cytosol was collected using the / mitochondria fraction kit), and cleavage caspase-3 was measured by Western blot by dissolving the whole protein.

As a result, as shown in Figure 15, by the pre-treatment of the phalanx extract was suppressed the disruption of mitochondrial membrane potential by 6-OHDA, thereby suppressing the increase in cytochrome C concentration in the cytoplasm to confirm the anti-self killing effect. In addition, the phalanx extract inhibited caspase-3 activity activated by 6-OHDA at 0.1-10 μg / mL concentration pretreatment and cleavage caspase-3 foam was also inhibited (FIG. 15).

< Experimental Example  5> In vivo In vivo )in Phalanges  Extract Anti-Parkinson's disease  Check the effect

<5-1> Cytotoxicity Test

Sprague is our rats to 1 × 10 ⁵ (Orient Bio, Seoul, Korea) for 14 days after MCA areas but isolated from fetal, then decomposed mechanically obtain the cells, poly -L- pre-coated 24-well flight as lysine Six days were incubated after inoculating cells with / well. Subsequently, after treating the phalanx extract dissolved in 0.1% DMSO / FBS-free medium at 0.01-10 ㎍ / mL, it acts on dopamine neurons and affects the mitochondria complex I channel, resulting in cytotoxicity. MPP + (10 μm), which is a representative substance, was treated and further incubated for 24 hours. In addition, in case of 6-OHDA toxicity, the phalanx extract was treated with 0.01-1 μg / mL 6 hours later, followed by 6-OHDA (10 μm) treatment for 18 hours, and after fixation, rabbit anti-tyrosine hydroxyxa TH-positive cells stained with rabbit anti-tyrosine hydroxylase (TH) were identified on a 100-fold microscope.

As a result, as shown in FIG. 16, the TH-positive cell number decreased by 6-OHDA and MPP + was treated at 10 μg / mL in 6-OHDA toxicity, and 10 or 100 μg / mL in MPP + toxicity by pregelatin extract. It was confirmed that the protective effect at.

<5-2> rotation inspection ( rotation test )

Sprag Dowley rats (Orient Bio, Seoul) were bred for one week, and 6-OHDA was injected into the medial forebrain bundle (MFB) using strotaxic. Thereafter, the phalanx extract was orally administered at 100 mg / kg, and a rotational test induced by apomorphine was performed on days 7 and 14.

As a result, as shown in Figure 17, after measuring the rotational speed per minute after 6-OHDA induced toxicity was confirmed that the amount of rotation of the phalanx extract 100 mg / kg administration group compared to the toxic group 7 days and 14 days (Fig. 17).

< Experimental Example  6> in vitro heartache and Phalanges  Mixed extract Anti-Alzheimer's  Check the effect

The present inventors performed the following experiment to confirm the anti-Alzheimer's effect of the mixture on the heart and extract extracts. In addition, the efficacy of the mixture in the heart beat and phalanges extract was compared with the efficacy of the extracts of each of the heart beat and phalanges.

<6-1> Hippocampus  Cytoprotective effect

In order to confirm the cytoprotective effect of the mixture of the heart and the phalanges, the hippocampus was isolated from the 18-day-old fetus of Sprague Dawley rats (Orient Bio, Seoul), and then mechanically decomposed to obtain cells. Cells were seeded at 1 × 10 ⁴ / well in 96 well flights pre-coated with -L-lysine and cultured for 7 days. Subsequently, after 30 minutes of treatment with the heart extract or the phalanx extract, or a mixture thereof, in a 0.1% DMSO / B27 free neuronal base medium, the incubation was performed for 24 hours with beta amyloid (Abeta, 8 μg) or non-treatment. After 3 hours of 1 mg / mL MTT treatment, formazan was dissolved in DMSO and absorbance was measured at 570 nm.

As a result, as shown in Figure 18, when treated with the heart extract or phalanx extract, or a mixture thereof alone in the hippocampal cells, the cell viability was affected or confirmed an increase in the cell, the heart extract or phalanges extract, or its When the mixture was pretreated, it was confirmed that the protective effect against Abeta toxicity (Fig. 18).

< Manufacturing example  1> Preparation of Pharmaceutical Formulations

<1-1> Sanje  Produce

Example 1-1 2 g of lettuce extract

Lactose 1 g

The above components were mixed and packed in airtight bags to prepare powders.

<1-2> Preparation of tablets

100 mg of lettuce extract of Example <1-1>

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

&Lt; 1-3 > Preparation of capsules

100 mg of a mixture of the heartache and phalanges of Example <1-3>

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

&Lt; 1-4 >

Phalanx extract 1g of Example <1-2>

Lactose 1.5 g

1 g of glycerin

Xylitol 0.5 g

After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.

<1-5> Preparation of granules

150 mg of phalanges extract of Example <1-2>

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

After mixing the above components, 100 mg of 30% ethanol was added and the mixture was dried at 60 캜 to form granules, which were then filled in a capsule.

< Manufacturing example  2> Manufacturing of food

Foods containing the heart beat extract of the present invention was prepared as follows.

<2-1> Production of flour food

0.5-5.0 parts by weight of the lettuce extract of Example <1-1> was added to the flour, and bread, cake, cookies, crackers and noodles were prepared using this mixture.

<2-2> soup  And juicy ( gravies Manufacturing

0.1 ~ 5.0 parts by weight of the heart extract extract of Example <1-1> was added to soups and broths to prepare meat products for health promotion, soups of noodles, and broths.

<2-3> Ground Beef  Produce

10 parts by weight of the lettuce extract of Example <1-1> was added to ground beef to prepare a ground beef for health promotion.

<2-4> Dairy products ( dairy products Manufacturing

5-10 parts by weight of the heart extract extract of Example <1-1> was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<2-5> Solar  Produce

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh.

Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer.

The concentrated heart extract of Example <1-1> was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by drying with a sprayer and a hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder.

The cereals, seeds and the extract of Example 1-1 prepared above were formulated in the following ratios:

(30 parts by weight of brown rice, 15 parts by weight of yulmu, 20 parts by weight of barley)

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)

Heartworm extract (3 parts by weight) of Example <1-1>,

(0.5 parts by weight), and

Rhubarb (0.5 parts by weight).

< Manufacturing example  3> Manufacturing of beverage

<3-1> Health drink  Produce

Homogeneously blends subsidiary materials such as liquid fructose (0.5%), oligosaccharides (2%), sugar (2%), salt (0.5%), water (75%) and 5 g of phalanges extract of Example <1-2> After the instant sterilization it was prepared by packaging in a small packaging container such as glass bottles, plastic bottles.

<3-2> Preparation of vegetable juice

5 g of phalanges extract of Example <1-2> was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice.

<3-3> Production of fruit juice

1 g of phalanges extract of Example <1-2> was added to 1,000 ml of apple or grape juice to prepare a fruit juice.

Claims (4)

Jigolpi (scientific name: Lycium chinense ) Parkinson's disease (Parkinson's disease) prevention and treatment pharmaceutical composition containing the extract as an active ingredient.
Heartache (scientific name: Morus alba ) and Alzheimer's prevention and treatment containing a mixed extract of the phalanges as an active ingredient.
Parkinson's disease prevention and improvement health functional food containing the phalanges extract as an active ingredient.
Health functional food for the prevention and improvement of Alzheimer's containing mixed extract of lettuce and phalanges as an active ingredient.

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