CN107158012A - Application of the Morinda officinalis FOS extract in preventing and treating Alzheimer disease drug, health products or food is prepared - Google Patents

Abstract

The invention discloses the application of the fructooligosaccharide extract from Morinda officinalis in the preparation of anti-Alzheimer's medicine, health care products or food. The present invention has found through animal experiments in vivo that the fructooligosaccharide extract from Morinda officinalis can significantly improve the Aβ _1‑42 and D‑galactose induced the learning and memory ability of model rats with simulated senile dementia; the animal acute toxicity experiment confirmed that the safe dose range of Morinda officinalis fructooligosaccharide extract is relatively large, and it can be used as a drug for the prevention and treatment of Alzheimer's disease , health products and food and other fields.

Description

The extract of Morinda officinalis fructooligosaccharides is used in the preparation of anti-Alzheimer's disease medicine, health care food or food applications

technical field

The invention relates to a new application of the fructooligosaccharide extract from Morinda officinalis, in particular to the application of the fructooligosaccharide extract from Morinda officinalis in the preparation of anti-Alzheimer's medicine, health products or food.

Background technique

Alzheimer's disease (AD) is a neurodegenerative disease with insidious onset and progressive development. Clinically, it is characterized by comprehensive dementia such as memory impairment, aphasia, apraxia, agnosia, impairment of visuospatial skills, executive dysfunction, and personality and behavior changes. The etiology is still unknown. The etiology and pathogenesis of senile dementia are still unclear so far, and there may be multiple factors involved, such as aging, genetics, metabolism, history of head trauma, estrogen deficiency, etc.

Studies have found that the human gut has a very large number of neurons, which is comparable to the brain. The neurotransmitters and metabolic pathways used by intestinal neurons and brain neurons are almost the same. There is frequent communication from neurons in the gut to neurons in the brain, more than the exchange of information from the brain to the gut. The communication between intestinal microorganisms and the brain is very frequent. The academic community believes that a "bacteria-gut-brain axis" is formed between the gut and the brain. The gut-brain can affect the brain, and the brain can also affect the gut-brain. There is a two-way communication between them. They interact mainly through endocrine pathways, neural pathways, immune pathways and metabolic pathways. Infection during pregnancy, pollutants, pathogens, diet, inflammation and other factors can affect the communication between them. We can intervene through probiotics, diet, and fecal transplants.

Therefore, it is a new way to find foods, prebiotics, probiotics, plant extracts or drugs that can affect the structure or ecology of intestinal microorganisms for the treatment of neurodegenerative diseases.

The traditional Chinese medicine Morinda officinalis is an important medicine for invigorating the kidney and yang. Previous studies have shown that it is rich in oligosaccharides, and has the functions of nourishing kidney and yang, anti-depression, anti-osteoporosis, and improving learning and memory ability, immunity and fatigue resistance. At present, the research on Morinda officinalis fructooligosaccharides at home and abroad mainly focuses on antipsychotic agents, immune adjuvants, etc., but there are few studies on the application of Morinda officinalis fructooligosaccharides extracts in the prevention and treatment of Alzheimer's disease. .

Contents of the invention

The purpose of the present invention is to provide the application of Morinda Citrifolia fructooligosaccharide extract in the preparation of anti-Alzheimer's disease medicine, health products or food.

Preferably, the safe dose range of the extract of Morinda officinalis fructooligosaccharides is 30 mg/kg-500 mg/kg.

Preferably, the preparation method of the Morinda officinalis fructooligosaccharide extract comprises the following steps:

(1) Remove impurity from the fresh medical material of Morinda officinalis Morinda officinalis, and properly pulverize;

(2) adding distilled water for extraction, and concentrating the extract under reduced pressure to obtain a concentrated solution;

(3) adding ethanol to the concentrated solution for precipitation, redissolving the precipitate in water, adding ethanol for reprecipitation, and obtaining the crude product of Morinda officinalis fructooligosaccharide;

(4) Redissolve the crude product of Morinda officinalis fructooligosaccharides in water, elute with anion exchange resin, collect the eluent, concentrate the eluent under reduced pressure, add ethanol to precipitate, let stand, filter with suction, and dry to obtain Morinda officinalis oligosaccharide extract.

Preferably, the extraction method of the step (2) is one or more of heating reflux, hot soaking or percolation.

Preferably, the specific steps of the step (2) are: add 8-10 times the amount of distilled water, boil for 1-2h, pass

Filter, add 8-10 times the amount of distilled water to the filter residue, boil for 1-2h, filter, combine the filtrates, and concentrate under reduced pressure to obtain a concentrated solution.

Preferably, the specific steps of the step (3) are: add ethanol to the concentrated solution, adjust the ethanol concentration to 90%, refrigerate, let stand for 2-24 hours, redissolve the precipitate in hot water, cool to room temperature, and then add Ethanol, adjust the ethanol concentration to 90%, refrigerate, let stand for 2-24h, and get precipitate, which is the crude product of Morinda officinalis fructooligosaccharide.

In the present invention, the operations of redissolving the precipitate in water and adding alcohol to precipitate can be repeated many times so as to better remove alcohol-soluble impurities.

Preferably, the specific steps of the step (4) are: redissolving the crude fructooligosaccharide of Morinda officinalis in hot water, cooling to 50°C, adjusting the pH to 4.5-6.5, passing the solution through D-900 anion exchange resin, Continuously elute at a flow rate of 1.0-1.5BV/h, collect the eluent, concentrate under reduced pressure at 50-70°C to a relative density of 1kg: 200ml volume, add ethanol to the concentrated solution, adjust the ethanol concentration to 90%, refrigerate, and statically Set aside for 2-24h, stand still, filter with suction, and dry to obtain the oligosaccharide extract of Morinda officinalis.

The Morinda officinalis fructooligosaccharide extract obtained by the above-mentioned preparation method of the present invention is detected by combining chemical identification, HPLC, LC-MS and other technologies, and the results show that it mainly contains Morinda officinalis, Nessose, 1F-fructofuranosyl Nessose, inulin (ie (2→1) fructofuranosyl sucrose) type hexasaccharides and heptasaccharides and other oligosaccharides, the yield is greater than 30%.

The present invention also provides a medicine for preventing and treating Alzheimer's disease, which comprises the above-mentioned Morinda Citrifolia fructooligosaccharide extract and a pharmaceutically acceptable carrier.

The dosage form of the medicine of the present invention can be tablet, granule, powder, capsule or oral liquid, etc., and its preparation method can be prepared by a conventional method.

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

The present invention has found through in vivo animal experiments that the extract of Morinda officinalis fructooligosaccharides can significantly improve the learning and memory ability of model rats with simulated senile dementia induced by Aβ1-42 and D-galactose; The dosage range is large, and it can be used in the fields of prevention and treatment of Alzheimer's disease drugs, health products and food.

Description of drawings

Figure 1 is the HE staining diagram of the intestinal and brain pathological tissues of the aging model rats induced by D-galactose by the extract of Morinda officinalis fructooligosaccharides.

Figure 2 is the effect of Morinda officinalis fructooligosaccharide extract on Effects on intestinal, heart, and brain pathological tissues of rats with dementia, as well as in brain tissue HE staining and immunohistochemical diagrams of the influence of Tau protein expression level.

detailed description

The present invention will be further described below through specific embodiments. The following examples are specific embodiments of the present invention, but the embodiments of the present invention are not limited by the following examples.

The Morinda officinalis of the present invention is collected from the Morinda officinalis planting base in Gaoliang Town, Deqing City, Guangdong Province, and is identified as the dried root of Morinda officinalis of the Rubiaceae plant. Cylindrical form, slightly curved, varying in length, 1-2 cm in diameter, grayish-yellow or grayish-yellow-brown surface, some with purple tinge, with longitudinal wrinkles and deep horizontal lines, some constricted or The bark is cut off horizontally to expose the xylem, shaped like chicken intestines. The quality is tough, the broken surface is uneven, the thickness of the skin is 5-7mm, lavender, and the diameter of the xylem is 2-4mm. Gas micro, bitter, slightly astringent.

Embodiment 1: Preparation of Morinda officinalis fructooligosaccharide extract

Specifically include the following steps:

Remove the impurities of Morinda officinalis fresh medicinal material, crush it properly; add 8 times the amount of distilled water, boil for 2 hours, filter,

Add 8 times the amount of distilled water to the filter residue, boil for 2 hours, filter, combine the filtrates, concentrate under reduced pressure to obtain a concentrated solution; add ethanol to the concentrated solution, adjust the concentration of ethanol to 90%, refrigerate, stand for 24 hours, and redissolve the precipitate in hot water water, cooled to room temperature, then added ethanol, adjusted the ethanol concentration to 90%, refrigerated, left standing for 24 hours, and precipitated, which was the crude product of Morinda officinalis fructooligosaccharide; redissolve the crude product of Morinda officinalis fructooligosaccharide in hot water, Cool to 50°C, adjust the pH to 5.0, pass the solution through D-900 anion exchange resin, elute continuously at a flow rate of 1.0-1.5BV/h, collect the eluate, concentrate under reduced pressure at 50-70°C to a relative density of 1kg: 200ml volume, add ethanol to the concentrated solution, adjust the ethanol concentration to 90%, refrigerate, stand still for 2-24h, stand still, filter with suction, and dry to obtain Morinda officinalis oligosaccharide extract.

The above-mentioned Morinda officinalis extract (OMO) was subjected to a drug efficacy test:

Experiment 1: Morinda officinalis fructooligosaccharide extract improves learning and memory ability of D-galactose-induced dementia model rats

SPF grade SD rats were used for the experiment, and the model was established by long-term intraperitoneal injection of D-galactose to prepare the senile dementia model. After modeling, the normal control group and the model group were intragastrically administered with 2 mL/d normal saline, and the OMO low-dose group (50 mg/kg/d) and the OMO high-dose group (100 mg/kg/d) were administered with their respective dosages. Oral administration of Morinda officinalis fructo-oligosaccharide extract OMO for 8 weeks in each group. After 8 weeks, the spatial learning and memory ability of the rats was evaluated by Morris water maze (MWM). The results are shown in Table 1, Table 2, and Figure 1.

Table 1. The effect of Morinda officinalis oligosaccharide extract on the latent period of water maze test in D-galactose-induced aging model rats (n=6)

Table 2. The effect of Morinda officinalis oligosaccharides extract on the water maze test platform stage of D-galactose-induced aging model rats (n=6)

It can be seen from Table 1 that at different test time points of the incubation period test, compared with the normal group, the incubation period of the model group was significantly prolonged, and the difference was statistically significant (P<0.05), suggesting that the experimental model was successfully prepared. Compared with the model group, the incubation period of each administration group was significantly shortened, and the high-dose OMO group was better than the low-dose OMO group (P<0.05).

It can be seen from Table 2 that the swimming time of rats in the normal group was in the third quadrant (that is, the quadrant where the platform was originally located). Longer than other quadrants, the difference is significant, indicating that the rats have formed a memory for the platform. Compared with the normal group, the swimming time of the model group in the third quadrant significantly shorter, indicating that D-galactose can impair the memory function of rats; compared with the model group, the swimming time of the OMO high-dose group in the third quadrant Significantly prolonged (P<0.05), suggesting that Morinda officinalis fructooligosaccharide extract has the effect of improving D-galactose-induced memory dysfunction in rats.

It can be seen from Figure 1 that the small intestinal cells (Intestin) in the model group were shrunken and irregular, while the small intestinal cells in each administration group were smoother, regular in shape and plump. In the brain stem (Brainstem), the number of inflammatory cells in the model group was significantly more than that in the administration groups, suggesting that the administration group can reduce the inflammatory response of cells. In the cerebellum, compared with the model group, the number of cells in each administration group increased significantly.

Therefore, it can be seen from the above that the extract of Morinda officinalis fructooligosaccharides can improve the learning and memory ability of D-galactose-induced senile dementia model rats, and reduce tissue inflammation.

Experimental example 2: Improvement of Morinda officinalis fructooligosaccharide extract Inducing the learning and memory ability of simulated senile dementia model rats

SPF grade SD rats were used for the test, and bilateral intrahippocampal injection was used Modeling by method to prepare senile dementia model. After modeling, the normal control group and the model group were intragastrically administered with 2 mL/d normal saline, and the OMO low-dose group (50 mg/kg/d) and the OMO high-dose group (100 mg/kg/d) were administered with their respective dosages. Oral administration of Morinda officinalis extract OMO for 4 weeks. After 4 weeks, the spatial learning and memory ability of the rats was evaluated by Morris water maze (MWM). The results are shown in Table 3, Table 4, and Figure 2.

Table 3. Morinda officinalis oligosaccharide extract pair Effect of water maze test latency on rats with dementia (n=6)

Table 4. Morinda officinalis oligosaccharide extract pair The impact results of the water maze test platform period in the dementia model (n=6)

It can be seen from Table 3 that at different test time points of the incubation period test, compared with the normal group, the incubation period of the model group was significantly prolonged, and the difference was statistically significant (P<0.05), suggesting that the experimental model was successfully prepared. Compared with the model group, the incubation period of each administration group was significantly shortened, and the high-dose OMO group was better than the low-dose OMO group.

As can be seen from Table 4, the swimming time in the third quadrant (i.e. the quadrant where the platform was originally located), compared with the model group, the swimming time of each administration group was significantly prolonged (P<0.05), suggesting that Morinda officinalis oligosaccharide extract It can improve the learning and memory ability of Aβ1-42 induced senile dementia model rats.

It can be seen from Figure 2 that compared with the model group, in each administration group, the cells of the small intestine (Intestin) were more bright and plump, the gaps between the cells of the heart (Heart) became smaller, and the number of cells of the brain (Brain) increased significantly. and Tau protein expression decreased, suggesting that Morinda officinalis oligosaccharide extract can improve the function and morphology of rat tissue cells, and significantly reduce Alzheimer's disease-related proteins and Tau expression.

Therefore, it can be seen from the above that the oligosaccharide extract of Morinda officinalis can improve the learning and memory ability of Aβ1-42 induced senile dementia model rats, reduce and Tau protein expression.

Safety Evaluation:

Toxicity test, according to the "Technical Guidelines for Safety Evaluation of Traditional Chinese Medicine and Natural Medicines", the dosage range of Morinda officinalis fructooligosaccharide extract is 30mg/kg-500mg/kg. The maximum dose of 500 mg/kg was given to Kunming mice by intragastric administration, and no animal died after continuous observation for 14 days. It shows that the safe dose range of Morinda officinalis fructooligosaccharide extract is relatively large, and the medicine is safe.

Application example: Preparation of granules containing Morinda officinalis fructooligosaccharide extract

Active ingredients: 65 parts of Morinda officinalis fructooligosaccharide extract;

Excipients: 25 parts of pregelatinized starch, 4 parts of micronized silica gel, 6 parts of magnesium stearate.

Preparation process: put Morinda officinalis fructooligosaccharide extract, pregelatinized starch, micropowder silica gel and magnesium stearate into a mixer and mix evenly.

The above mixture is filled into capsules by a fully automatic capsule filling agent to obtain the product.

Claims (9)

1. The application of Morinda officinalis fructooligosaccharide extract in the preparation of anti-Alzheimer's disease medicine, health products or food. 2. the application of Morinda officinalis fructo-oligosaccharide extract according to claim 1 in the preparation and prevention and treatment of Alzheimer's disease medicine, health product or food, is characterized in that, the fructo-oligosaccharide extract of Morinda officinalis root The safe dosage range is 30mg/kg-500mg/kg. 3. the application of Morinda officinalis fructo-oligosaccharide extract according to claim 1 in the preparation and prevention and treatment of Alzheimer's disease medicine, health products or food, is characterized in that, the fructo-oligosaccharide extract of Morinda officinalis root The preparation method comprises the following steps: (1) Remove impurities from the fresh medicinal material of Morinda officinalis and properly crush it; (2) adding distilled water for extraction, and concentrating the extract under reduced pressure to obtain a concentrate; (3) Precipitate the concentrated solution with ethanol, redissolve the precipitate in water, add ethanol to reprecipitate, and obtain the crude fructooligosaccharide of Morinda officinalis; (4) Redissolve the crude product of Morinda officinalis fructooligosaccharides in water, elute with anion exchange resin, collect the eluent, concentrate the eluent under reduced pressure, add ethanol to precipitate, let stand, filter with suction, and dry to obtain Morinda officinalis oligosaccharide extract. 4. The application of Morinda officinalis fructo-oligosaccharide extract according to claim 1 in the preparation of anti-Alzheimer's disease medicine, health products or food, characterized in that the extraction method in the step (2) is One or more of heating reflux, hot dipping or percolation. 5. The application of Morinda officinalis fructooligosaccharide extract according to claim 4 in the preparation of anti-Alzheimer's disease medicine, health products or food, characterized in that, the specific steps of the step (2) are: Add 8-10 times the amount of distilled water, boil for 1-2 hours, filter, add 8-10 times the amount of distilled water to the filter residue, boil for 1-2 hours, filter, combine the filtrates, and concentrate under reduced pressure to obtain a concentrated solution. 6. The application of Morinda officinalis fructooligosaccharide extract according to claim 1 in the preparation of anti-Alzheimer's disease medicine, health products or food, characterized in that, the specific steps of the step (3) are: Add ethanol to the concentrated solution, adjust the ethanol concentration to 90%, refrigerate, let stand for 2-24h, redissolve the precipitate in hot water, cool to room temperature, then add ethanol, adjust the ethanol concentration to 90%, refrigerate, let stand for 2 -24h, a precipitate was obtained, which was the crude product of Morinda officinalis fructo-oligosaccharide. 7. The application of Morinda Citrifolia fructooligosaccharide extract in the preparation of anti-Alzheimer's disease medicine, health products or food according to claim 1, characterized in that, the specific steps of the step (4) are: Redissolve the crude fructooligosaccharide of Morinda officinalis in hot water, cool to 50°C, adjust the pH to 4.5-6.5, pass the solution through D-900 anion exchange resin, and continuously elute at a flow rate of 1.0-1.5BV/h, collect and wash Remove liquid, concentrate under reduced pressure at 50-70°C to a relative density of 1kg: 200ml volume, add ethanol to the concentrated solution, adjust the ethanol concentration to 90%, refrigerate, stand still for 2-24h, stand still, suction filter, dry, that is Morinda officinalis oligosaccharide extract. 8. A medicine for preventing and treating Alzheimer's disease, comprising Morinda Citrifolia fructooligosaccharide extract and a pharmaceutically acceptable carrier. 9. The drug for preventing and treating Alzheimer's disease according to claim 8, characterized in that, the dosage form of the drug is tablet, granule, powder, capsule or oral liquid.