WO2005077381A1

WO2005077381A1 - Use of ginsenoside re for preparation of medicaments for treating dysmnesia

Info

Publication number: WO2005077381A1
Application number: PCT/CN2004/000083
Authority: WO
Inventors: Pingya Li; Juntian Zhang; Wenjie Zhao
Original assignee: Beijing Xinliheng Pharmaceutical Science & Technology Evelopment Co., Ltd.
Priority date: 2004-01-29
Filing date: 2004-01-29
Publication date: 2005-08-25

Abstract

Use of ginsenoside re for preparation of medicaments for treating dysmnesia, dysmnesia, improving ß-amyloid peptide induced spatial identification disease, and promoting the basic synaptic transmission, and facilitating the formation of LTP.

Description

Application of ginsenoside Re in preparing medicine for treating memory disorder Technical field

The present invention relates to the application of ginsenoside Re as a medicine for treating memory disorders. Specifically, the ginsenoside Re can not only improve memory disorders caused by chemical drugs (scopolamine, etc.), but also improve natural aging. A β and other memory disorders and enhance the role of basal synaptic transmission and promote LTP formation. ·

Background technique

The prior art has disclosed pharmaceutical uses of some types of ginsenosides, such as-

CN1300593A discloses a medical use "ocotillol F _u of the literature indicates, ocotillol

F _n can significantly inhibit memory impairment caused by morphine and has a strong effect on improving learning and memory impairment in normal animals and animals.

CN1092203C discloses "the extraction process of ginsenoside Re and its new medicinal use. The document indicates that ginsenoside Re has a wide range of applications in the preparation of medicines for treating heart rate irregularities, myocardial ischemia and reperfusion injury.

CN1092204C discloses "a semi-synthetic method of 20 (S) -ginsenoside Rg ₃ and its pharmaceutical use", the document states that 20 (S) -ginsenoside Rg _{3 is} particularly suitable for the preparation and treatment of lung cancer, melanoma, and S ₁₈ o sarcoma , Colon cancer and antiviral drugs.

CN1331698A discloses a "brain cell or nerve cell protective agent containing ginsenoside Rt ^", and the article states that a preparation administering ginsenoside or a salt thereof as a cytoprotective agent is particularly suitable for the treatment, prevention, or treatment of brain and neurological diseases.

CN1309969A discloses "the application of ginsenoside Rg ₂ in the preparation of medicines for the treatment of cardiac and cerebrovascular diseases". The document states that the extraction and isolation of ginsenoside Rg ₂ from stems and leaves of ginseng are effective against various shocks, heart failure, myocardial ischemia and Cerebral ischemia has obvious therapeutic effect, its effect is better than the known Shenmai injection; it is used to treat diseases such as shock, heart failure, coronary heart disease, cerebral infarction and senile dementia.

However, none of the above public literatures specifically relates to the indications described in the present invention or merely propose concepts without specific experimental parameters and effect parameters. Summary of the invention

The purpose of the present invention is to realize the application of ginsenoside Re in medicine. In particular, the purpose of the present invention is to realize the preparation of ginsenoside Re to improve memory impairment caused by natural aging, A β, etc., and enhance basal synaptic transmission, and promote LTP. Application of the formed drug.

When the present invention is used to prepare a medicament for treating senile dementia or memory disorder, the oral or parenteral administration is safe. In the case of oral administration, it can be administered in any conventional form, such as powders, granules, tablets, capsules, pills, drip pills, soft capsules, flotation agents, oral solutions, suspensions, syrups, buccal tablets, sprays When the drug is administered orally, it can be in any conventional form, such as suppositories, injections (intravenous, intramuscular), ointments, inhalants, and the like.

The medicine for treating senile dementia and memory disorder prepared by the present invention may be composed of solid or liquid excipients. The solid or liquid excipients used herein are well known in the art. Here are some specific examples. Solid The excipients of the preparation are lactose, starch, dextrin, calcium carbonate, synthetic or natural aluminum sulfate, magnesium chloride, magnesium stearate, sodium bicarbonate, dry yeast, etc .; the excipients of the liquid preparation are water, glycerol, propylene glycol , Monosyrup, ethanol, ethylene glycol, polyethylene glycol, sorbitol, etc .; excipients for ointments can use fatty oils, hydrous lanolin, petrolatum, glycerin, beeswax, paraffin, liquid paraffin, resin, advanced wax And other combinations of hydrophobic or hydrophilic agents.

The dosage taken by the present invention is: 12.5-50mg / kg, and the preferred dosage is 25-50mg / k; taken 1 to 3 times daily.

Since the extraction process of ginsenoside Re and some other pharmaceutical uses have been disclosed in the prior art, and the main technical content of the present invention is to verify the effect of ginsenoside Re on the indications described in the present invention, therefore, the present invention For the extraction process of ginsenoside Re, please refer to the content disclosed in CN1092203C. In the present invention, various behavioral and electrophysiological methods commonly used in the world are used to observe the effect of ginsenoside Re on memory improvement in adult and elderly rats and various memory impairment models, and the electrophysiological method is used to observe Re The effects of long-term synapse enhancement (LTP) on synapses can be seen in the following experimental examples. Through the experimental examples, the present invention can be further understood.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram of a mouse water maze device used in the present invention.

The present invention can be clearly understood through the accompanying drawings of the specification and the specific content of the present invention. Experimental example 1

This experimental example involves the study of ginsenoside Re promoting the learning and memory of naturally aging mice and rats.

(1) Effect of ginsenoside Re on memory dysfunction caused by natural aging in a mouse water maze: Take 64 SPF Kunming male males, weighing 50 to 2 g, randomly divided into 5 groups, normal elderly control group, ginsenoside Re group (12.5mg / kg, 25mg / kg and 50mg / kg), Naofukang group (500mg / kg), plus a normal young control group of SPF Kunming mice. Normal elderly and young controls received water. The rest were orally administered, and mice were trained on the water maze from day 11 to day 15. The schematic diagram of the maze is shown in FIG. 1.

It can be seen from Figure 1 that the water maze consists of a black plexiglass rectangular parallelepiped (73 X42 X20cm), which has several partitions to form the maze: where 1, 2, 3, and 4 are blind ends, and a set of steps at the corner of the maze are the end points . During the experiment, water was poured into the maze with a water depth of 10 cm and a water temperature of 24 ° C and 1 ° C.

On the first day of training, the channel between points A and B was blocked with a partition. The mouse was first placed on a step for 10 seconds to make it aware of the existence of this safe area, and then the mouse was placed at the starting point A and let it Swim freely, record the number of times it enters blind end 1 (ie the number of errors) and the time to climb the safety steps (ie the incubation period). If no steps are found within 2 minutes of training time, they will be guided to the safety steps and their latency period Recorded as 2min. Take a break after the first training and train the second time in the same way. Train the other mice in turn.

On the second day of training, place the partition at point B, and the mouse's swimming starting point is point B. Record the number of times that the mouse entered the blind end 1, 2 and 3 within 2 minutes, and the incubation period of climbing the safety steps. Mice were trained twice in a row. If no step can be found within 2 minutes, it will be guided to the end point, and its incubation period is recorded as 2 minutes.

On the third to fourth day of training, the swimming starting point of the mouse is point C, and the number of errors of the mouse entering the blind end 1, 2, 3, 4 and the incubation period of finding a safety step within 2 minutes are recorded. Each mouse is trained twice in a row.

Table 1 Effect of ginsenoside Re on memory dysfunction caused by natural aging in mouse water maze

(2) The role of ginsenoside Re in preventing dark memory caused by natural aging in mice 64 elderly SPF Kunming mice, weighing 50 ± 2g, male, were randomly divided into 5 groups, the normal elderly control group, the ginsenoside Re group (12.5mg / kg, 25mg / kg, and 50mg / kg), and the brain rehabilitation group (500mg / kg), plus a normal young control group. Normal elderly and young controls received water. The rest were administered by oral gavage, and mice were trained to avoid darkness on the 16th and 17th days after administration.

The experimental device is a conditional reflection box, which is divided into two parts, light and dark. There is a small hole in the two chambers to allow mice to pass freely. The bottom of the box has a copper grid. The copper grid at the bottom of the dark room is connected to 36V voltage. During the experiment, the mice were placed in a bright room with their backs facing the dark room. Because the mice have the habit of drilling darkness and drilling holes, they will drill into the dark room multiple times during the experimental training. You will be shocked. This behavior is an error, and the normal response is to try to exit the dark room immediately. When training on the first day, record the number of times the mice entered the darkroom within 3 minutes and the time when they first entered the darkroom. The experiment was performed after 24 hours. Observe the time when the mouse first enters the dark box within 5 minutes, which is the incubation period, and the number of times the mouse enters the dark box is the number of errors.

It can be seen from Table 2 that low, medium and high doses of ginsenoside Re have significant differences in latency and number of errors, and high doses of ginsenoside Re are more significant.

Table 2 Effect of ginsenoside Re against memory dysfunction caused by natural aging in the dark avoidance experiment in mice

(3) Effect of ginsenoside Re on memory dysfunction caused by natural aging in rat Morris water maze test

Fifty-six elderly SD male rats were randomly divided into five groups, the normal elderly control group, the ginsenoside Re group (6.25mg / kg, 12.5mg / kg, and 25mg / kg), and the brain rehabilitation group (250mg / kg), And add a normal young control group. Normal elderly and young controls received water. The rest were administered by oral gavage, and rats were trained on the water maze from day 15 to day 19.

The Morris water maze consists of a cylindrical pool made of stainless steel and a movable platform. A video camera is connected over the pool to the monitor and computer. When the set training time has expired or the animal has climbed onto the platform, the computer stops tracking or recording, and records the swimming trajectory and automatically calculates the distance the animal has traveled in the pool. The time required to find the platform (ie the incubation period) ) And initial angle (rats immediately after entering the water The angle between the long axis of the body and the line connecting the platform at the water entry point).

Fill the pool with clear water in advance, and then add 1 kg of fresh milk powder in an aqueous solution to make the pool water opaque milky white. Then fill the pool with water to make the water surface 1cm higher than the platform, and control the water humidity at 24 ± 0.5 ° C. Place the platform in a quadrant In the middle, in the other three quadrants, one of the water entry points is selected. The experiment is trained twice a day, each time a different water entry point is used, and the training is performed for 1 min each time. If the animal has not found a platform within lmin, take the animal to the platform and let it stay on it for 15s. If the rat finds the platform within lmin, also let it stay on the platform for 15s before finishing a training session. After five days of training in this way, the incubation period and the distance traveled by each animal were recorded separately. On the last day of training, remove the platform, each rat swims for 1 minute, and use a stopwatch to record the time that each rat first passed the platform, which is the incubation period, and record the passage of each rat within 1 min. The number of platforms and the time spent in the quadrant where the platform is located (the first quadrant). The shorter the incubation period of a rat and the larger the latter two indicators, the smarter the rat is.

Table 3 Ginsenoside Re in the five days before the rat Moms water maze test

Fight against memory dysfunction caused by natural aging

Incubation period Naofukang Ginsenoside Re Ginsenoside Re Ginsenoside Re Old mice Young mice

(Seconds) (250mg kg) 6.25mg / kg 12.5mg / kg 25mg / kg

(n = 12) (n = 14)

(n = H) (n = 9) (n = H) (n = 13)

D1 51.9 ± 15.6 44.6 ± 17.7 48.1 ± 20.7 57.2 ± 8.2 51.4 ± 16.2 55.2 + 9.4

D2 43.4 ± 22.0 25.0+ 19.9 ** 43.0 + 19.3 49.0 ± 17.8 47.4 ± 19.5 37.3 ± 21.0

D3 43.4 ± 23.9 20.0 ± 21.3 ** 31.5 + 20.4 * 36.3 ± 22.9 35.3 ± 25.9 32.2 ± 21.4

D4 27.8 ± 21.2 16.1 ± 14.5 * 24.6 ± 17.3 23.1 ± 20.6 26.4 + 20.9 16 · 8 ± 10.7 *

D5 25.9 ± 21.0 15.2 ± 16.6 * 28.5 ± 22.1 21.9 + 21.2 29.7 ± 22.7 16.5 ± 15.7 * Swimming distance Rehabilitation Ginsenoside Re Ginsenoside Re Ginsenoside Re Old mouse Young mouse

(cm) (250mg / kg) 6.25mg / kg 12.5mg / kg 25mg / kg

(n = 12) (n = 14)

(n = H) (n = 9) (η = Π) (n = 13)

D1 1139 ± 387 1054 ± 417 983 ± 454 992 ± 206 927 ± 369 * 930 ± 216 *

D2 936 ± 513 604 ± 483 994 ± 486 1053 ± 391 1078 ± 543 790 ± 448

D3 953 ± 564 498 ± 525 ** 671 soil 482 * 714 ± 477 749 ± 665 675 ± 463 *

D4 470 + 395 364 ± 302 453 ± 310 413 ± 316 500 ± 530 379 ± 298

D5 548 ± 503 454 ± 519 737 + 779 435 ± 389 632 ± 624 403 ± 337 Table 4 Ginsenoside Re

Effect on memory dysfunction caused by natural aging

Incubation period ^# : What is actually recorded here is the time that the rat swam across the platform for the first time within 1 minute after the platform is withdrawn, which is equivalent to the incubation period of five days before training.

The first quadrant time refers to the total time that the rats stay in the first quadrant when swimming, that is, the ratio of the time spent in the first quadrant to the total time.

In the above rat water maze experiment, compared with young rats, the aged rats were compared with young rats in terms of latency, swimming distances on the second and third days, and on the third day of training. There were significant differences in the initial angle on the last day and the number of passes on the last day of the training, indicating that this elderly rat model was established. In the brain rehabilitation group on the third and last day of training, the high-dose Re was on the fourth and fifth days of training in terms of latency, and in the brain rehabilitation group and the high-dose Re group were swimming on the third day of training In terms of the initial angle, the high dose of Re in the first quadrant residence time on the last day of training, and the medium dose of Re on the last day of training in terms of latency and number of steps were significant compared with older rats difference.

Experimental example 2

This experimental example concerns the improvement of ginsenoside Re on A β 25-35 in causing spatial discrimination disorders in rats.

The main pathological characteristics of senile dementia are the deposition of senile plaques, the formation of nerve fiber tangles and the loss of neurons, and the main clinical manifestations are memory and cognitive dysfunction. β-amyloid peptide (β-amyloid peptide, Αβ) (consisting of 1 to 40 amino acids) is an important component of senile plaque, which has a direct effect. After toxic effects. The peptide chain A β 25-35, which consists of 11 amino acids from 25 to 35 in A β, also has cohesive ability and neurotoxicity. We injected condensed A β 25- 35 in the lateral ventricle to cause spatial learning and memory impairment and observe Improvement effect of ginsenoside Re.

Male Wistar rats weighed 200 g ± 10 g. A β 25-35 was purchased from Sigma. The condensed A β 25-35 was prepared by dissolving 1 mg of A β 25- 35 in 3.14 ml of sterilized distilled water. The final concentration 3mmol 丄^-1 (15ιηιο 1/5 μ 1) sealed and placed in a 37 ° C incubator for 120h.

Forty-eight rats were divided into four groups, namely the surgical control group, the model group, the 25 mg / kg Re group, and the 12.5 mg / kg Re group. The animals were injected intraperitoneally with chloral hydrate (150 mg / kg) or urethane (750 mg / kg). After anesthesia, it was fixed on the stereotaxic instrument. After shaving and disinfecting the skin with 75% alcohol, the skull top skin was cut approximately 2cm in the front-to-back direction. Carefully peel off the subcutaneous tissue and periosteum to fully expose the skull. Use an electric drill to locate the coordinates Ao. _8, L _u position, the right side of the skull drilled holes with a diameter of 1.5mm, the removal of bone chips to prick dura carefully pointed tweezers, and then under stereotactic guide apparatus, the outer diameter of the steel pipes 0.8mm Insert it into the ventricle, inject 5 μ 1 ΑΡ 25—35 or an equivalent amount of sterile distilled water (control) slowly (3min) into the lateral ventricle for 2min, then withdraw a small steel tube and sprinkle a small amount of penicillin powder on the skull around the small hole. The next day, the stomach was administered once a day, and the Morris water maze training was performed for 14 days after continuous administration for 5 days. The platform was removed on the fifth day to record the time required for the animal to find the platform, that is, the incubation period, and the animal was placed on the platform after the platform was removed. Strategies to stay within the time limit to reach the platform and taken (linear, trend type, edge type, random type).

Morris water maze device and test see experimental example 1.

Table 5 Ginsenoside Re in Morris water maze test

Improving effect on spatial discrimination disorder caused by Α β 25- 35

The measurement result on the fifth day indicates that the animals stay in the platform quadrant after the platform is removed. Experimental example 3

This experimental example relates to the effect of ginsenoside Re against memory dysfunction caused by scopolamine in a mouse dark avoidance experiment, a mouse water maze, and a Morris water maze test.

The effect of ginsenoside Re on memory dysfunction caused by scopolamine in the dark avoidance experiment in mice is as follows: 72 SPF Kunming mice, weighing 20 ± 2g, male, were randomly divided into 6 groups, normal control group, scopolamine model group, Naofukang group, ginsenoside Re group (12.5mg / kg, 25mg / kg and 50mg / kg), oral administration for 9 days, normal control group and scopolamine model group, water administration, administration 9th and 10 夭 for dark avoidance training. The test method used is the same as before.

Referring to Table 6, it can be seen from the experimental results that both Naofukang and Ginsenoside Re have anti-scopolamine-induced memory impairment in mice, and the effect in the medium-dose group is more significant.

Table 6 Effect of ginsenoside Re on memory dysfunction caused by scopolamine in dark avoidance experiments in mice (n = 12)

Compared with scopolamine model group, * P <0.05, ** P <0.01

The effect of ginsenoside Re on memory dysfunction caused by scopolamine in the water labyrinth of mice is as follows: 72 SPF Kunming mice, weighing 20 ± 2g, male, were randomly divided into 6 groups, normal control group, scopolamine model group, The Naofukang group and the ginsenoside Re group (25mg / kg, 50mg / kg, and 100mg / kg) were orally administered with 17 mg of water, and the normal control group and the scopolamine model group were perfused with water. Mice water maze training was performed on days 14 to 18 of the administration. Except the normal control group, the other five groups received intraperitoneal injection of scopolamine (5mg / kg) every day during the first four days of training, and all animals were not injected with scopolamine five days before training. The test method is the same as before. The swimming starting point of the mice from the third to fifth days of training is point C, and the number of errors of the mice entering the blind end 1, 2, 3, 4 and the latency period to find a safety step within 2 minutes are recorded. Each mouse is trained twice.

As can be seen from Table 7, on the fourth day of training, only the normal control group and the scopolamine model group had significant differences in latency and number of errors, and the Naofukang group had only differences in latency compared with the model group. On the fifth day of training, there were significant differences in latency and number of errors between the normal control group and the Naofukang group and the scopolamine model group. Compared with the scopolamine model group, the low-dose ginsenoside group had an incubation period and number of errors. There are differences in the incubation period. It can be seen that the normal control group, the Naofukang group and the low and medium ginsenoside groups (25, 50mg / kg) have an anti-scopolamine effect in mice water maze experiments. .

Table 7 Effect of ginsenoside Re on memory dysfunction caused by scopolamine in mouse water maze experiment (n = 12)

Compared with scopolamine model group, * P <0.05, ** P <0.01

The Morris water maze test is as follows: 72 SPF Kunming mice, weighing 20 ± 2g, male, were randomly divided into 6 groups, normal control group, scopolamine model group, brain rehabilitation group, ginsenoside Re group (25mg / kg , 50 mg / kg and 100 mg / kg), orally administered for 17 days, the normal control group and the scopolamine model group were administered water, and mice were trained on the 14th to 18th days by water maze training. The other five groups except the normal control group received intraperitoneal injection of scopolamine (5mg / kg) 20 minutes before training every day for the first four days of training. All animals were not injected with scopolamine for the first five days of training. The test method is the same as before.

As can be seen from Table 8, on the fourth day of training, only the normal control group and the scopolamine model group have significant differences in incubation period and distance to seek a platform. The low-dose ginsenoside Re group is only in the There are different trends. On the fifth day of training, the normal control group differed in scopolamine model group in terms of incubation period and β distance, while the high-dose ginsenoside Re group had differences in latency compared with the scopolamine model group. The high-dose saponin group has a certain effect on the memory impairment caused by scopolamine in the Morris water maze experiment.

Table 8 Effect of ginsenoside Re on memory dysfunction caused by scopolamine in mouse Morris water maze experiment (n = 12)

Compared with scopolamine model group, * P <0.05, ** P <0.01

From this experimental example 3, the following conclusions can be drawn: mice were orally administered with ginsenoside Re at low doses (12.5mg / kg), medium doses (25mg / kg) and high doses (50mg / kg), avoiding darkness in mice In the experiment, it can obviously resist the memory acquisition disorder caused by scopolamine; in the mouse water maze experiment, the ginsenoside Re at a dose of 25, 50mg / kg has an effect against the memory acquisition disorder caused by scopolamine. In the mouse water maze experiment, 50mg / The kg dose of ginsenoside Re may have an effect against memory impairment caused by scopolamine.

Experimental example 4

This experimental example relates to the effect of ginsenoside Re on memory impairment caused by transient cerebral ischemia in mice, as follows: Kunming male mice, weighing 18-22 grams, were weighed, and pentobarbitone 60 mg / kg intraperitoneal anesthesia for surgery. The animals were randomly divided into 5 groups 24 hours after the operation, and the Re was administered orally and the doses were 6.25, 12.5, and 25 mg / kg body weight. The control group and the model group were given a solvent. Thereafter, it was administered once daily until the end of the behavioral test. After 48 hours, the common carotid arteries on both sides were completely blocked, and the blood supply was restored after the animals' righting reflex disappeared. Behavioral experiments were performed after 24 hours.

Models of transient cerebral ischemia were prepared and the animals were supine after anesthesia. Cut the skin about .7mm in the middle of the neck, separate the common carotid arteries on both sides, and place a white thread No. 0 under the left artery and a black thread No. 0 under the right artery. Blood flow. The two ends of each line are swallowed together, and the four terminals are passed through a section of PE50 plastic pipe with a length of about smm. The skin is penetrated from the middle of the neck and back, and the plastic is managed in the skin. Another black silk thread was passed through the annulus of the blood vessel through the black and white threads, and the two ends of the thread led out of the skin from the incision at the lower part of the neck, and the incision was sutured. During ischemia, gently press the skin on the back of the neck with a pair of tweezers, tighten the terminals of the four black and white wires, and fix the four wires to the skin with an arterial clip, resulting in cerebral ischemia. After the ischemia is over, loosen the arterial clamp and gently pull the sub-neck lead to fully restore the carotid artery to reperfusion.

Dark-proof experiment Before the experiment, mice were placed in a dark-proof box to acclimate to 3πύη. During the experiment, the mice were placed in the bright room with their backs facing away from the hole. The animals passed through the hole into the dark room and received electric shock. The mice were removed after 1 minute. After 24 hours, the test was repeated to record the incubation period of animals entering the dark room and the number of electric shocks within 5 minutes. '

Water maze experiment Record the time required for the animal to reach the target area from the starting area and the number of errors that the animal enters the blind end during this period. These two indicators are used as the learning and memory performance. The animals were trained once a day for 2 min each, for 4 consecutive days.

Data processing The data were expressed as mean ± s, and the t test was performed between groups.

Table 9 Effect of ginsenoside Re on learning and memory dysfunction caused by cerebral ischemia-reperfusion in dark avoidance experiment

Table 10 Effect of ginsenoside Re on learning and memory dysfunction caused by cerebral ischemia-reperfusion in water maze experiment

* P <0.05 ** P <0.01

In the dark avoidance test, compared with the model group, the sham operation group has behavioral indicators such as latency and number of errors. There are significant differences, and the model is established. Rel 2.5mg / kg, 25mg / kg can reduce the number of errors.

In the water maze test, the swimming incubation period on the fourth day was significantly shorter than that of the model group. Compared with the model group, the 25mg / kg group has a very significant difference. The incubation period of the 6.25mg / kg and 12.5mg / kg groups is shorter than that of the model group, but there is no significant difference.

Experimental example 5

This experimental example concerns the effect of ginsenoside Re on the synaptic transmission activity in the hippocampal dentate gyrus of anesthetized rats. The current research on nootropic drugs not only provides evidence of behavioral research, but also uses electrophysiology techniques to prove Its effect on synaptic plasticity is mainly to observe the effect on long-term synapse enhancement (LTP) of synapses, and it is considered as a cellular model of learning and memory of the central nervous system.

The specific experiment is that Wistar rats are anesthetized with 1.0% / kg 20% urethane and then fixed on a stereotaxic device. The positioning coordinates _{_{_{AP Q. 8, L u,}}} H 3. 5 a basket at planting tube in the lateral ventricle, the _{_{_{A0 3. 8, L 2.}}} 5, H at _{_3.5} implant recording electrodes in a hippocampal dentate gyrus granule cell layer, the _{_{_{AP 7. 5, L 4.}}} 2, H 3. 5 _4 .. A stimulation electrode was implanted in the perforant path (PP) of the entorhinal region. Adjust the position of the stimulating electrode to find the position where the maximum population spike amplitude (PSA) can be recorded under a fixed stimulus intensity, and it is fixed. The test stimulus is a square wave with a width of 0.2ms, with a frequency of 1 / 30Hz, and the stimulus intensity is 1/3-1/2 of the stimulus intensity required to cause the maximum PSA. Re was dissolved in 0.01% DMSO and diluted to different concentrations. It was administered through a basket implanted into the lateral ventricle, and the control group was given 0.01% DMSO. Record the PSA 30 minutes before the administration, and then slowly inject 5 μ1 of the test substance into the lateral ventricle within 5 minutes. Record the PSA within 60 minutes after the administration, and take three points of 15 minutes, 30 minutes, and 60 minutes for statistics. Take the average value of PSA before administration to represent the level of basal synaptic transmission before administration, and then use this average value to remove the PSA at 15min, 30min, and 60min after administration to obtain the relative value of PSA at each point (%). The mean and standard deviation were calculated within the group, and the t test was performed between the groups.

It can be seen from Table 11 that lateral ventricle injection of Re can enhance the synaptic transmission response of the hippocampal dentate gyrus. Re concentration of 10 · ⁵ Μ increase is not very significant, in 60min when reaching 135 ± 20.6%; with decreasing concentration of Re, an increase of PSA gradually increased, 10_ ⁶ M in 30min reached 138.9 ± 10.6% With 1 (T ⁷ M concentration, the largest increase, reached 138.4 ± 18.0% 15 minutes after administration, and continued to increase, reaching 189.6 ± 63.0% by 60 minutes. It can be regarded as the optimal dose; 10- ^S M PSA increased more than 1 (Γ ⁷ Μ decreased again, but also reached 152.2 ± 32.4% in 30 minutes. 1 (Τ ⁶ Μ, 1 (Γ ⁷ Μ, 1 (Τ ⁸ Μ) PSA increase in all three dose groups exceeded 30. Table 11 Effect of ginsenoside Re on hippocampal LTP formation in rats

P <0.05, * P <0.01, *** P <0.01 vs control

Long-term potentiation (LTP) of synaptic transmission is considered to be a cytological model of learning and memory of the central nervous system, becoming a cytological model of learning and memory, and an objective indicator of neuronal physiological activities in the process of learning and memory. This experiment found that the administration of ginsenoside Re at the doses of 10 ' ⁶ M, 1 (T ⁷ M, 10 · ⁸ M, lateral ventricle can enhance the synaptic transmission in the hippocampal dentate gyrus of anesthetized rats, and can form an LTP image. This result has enriched new content for the research on the nootropic effect of ginseng.

Ginsenoside Re is an important chemical component in ginseng. It can improve memory disorders caused by chemical drugs (scopolamine, etc.), and improve memory disorders caused by natural aging, AP, etc., and enhance basal synaptic transmission and promote LTP formation. It was found for the first time that it provided a sufficient experimental basis for developing ginsenoside Re into a nootropic drug.

detailed description

The method for preparing the medicine of the present invention is further illustrated by the following examples.

Example 1:

Capsules: Ginsenoside Rel80g, starch 20g, mixed hook, filled in No. 2 capsules, made into 1000 capsules, specifications: 0.2g / capsule. Adults take orally, 1 capsule / time, three times a day for one month.

Example 2:

Tablets: 180g total ginseng root saponin, 20g starch mixed hook, made into 1000 tablets, specifications: 0.2g / tablet; oral for adults, 1 tablet / time, 5 times a day, one course per month.

Claims

Claim

1. Application of ginsenoside Re in preparing medicine for treating senile dementia and memory disorder.

2. Application of a ginsenoside Re in the preparation of a medicine for improving the spatial discrimination disorder caused by β-amyloid peptide.

3. Application of a ginsenoside Re as a medicine for enhancing basal synaptic transmission and promoting LTP formation.

4. The use according to any one of claims 1 to 3, characterized in that the amount of the ginsenoside Re is 12.5-50 mg / kg.