JP2000229876A - A substance containing an active ingredient of Kinnasai extract - Google Patents

Abstract

(57) [Summary] [PROBLEMS] To provide an anti-dementia substance and an anti-stress agent. SOLUTION: An anti-dementia substance comprising, as an active ingredient, an extract obtained by extracting from the Japanese cabbage, and an anti-stress-acting substance comprising, as an active ingredient, the extract obtained from the Japanese cabbage.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a substance containing a golden soybean extract as an active ingredient.

[0002]

2. Description of the Related Art Golden needle vegetables are dried flowers of the buds of a lily plant, a hemlock, and are also called yellow flowers, and the dried buds are used as food.

[0003] It has been reported that golden sardines contain a large amount of melatonin-like active substances. Melatonin is a hormone secreted by the pineal gland, and it has been reported that melatonin-like substances have an effect of improving sleep disorders and are effective in eliminating jet lag. Activity has been noted.

[0004]

Aging is an unavoidable phenomenon for all living things, but in modern society, care for the elderly with dementia has become a major problem not only for individuals but also for the whole society. Therefore, development of an anti-dementia substance as a countermeasure against the aging phenomenon, particularly dementia, is strongly desired.

It has been studied for a long time that stress is involved in one of the major causes of modern diseases such as cancer and myocardial infarction, and it has a high anti-stress effect that removes this stress or suppresses the generation of stress. The development of drugs or foods has long been desired. The present applicant has discovered that the golden harvest has an anti-dementia effect and an anti-stress effect, and tried to commercialize this.

[0006]

Means for Solving the Problems The anti-dementia substance according to the first invention of the present application is characterized in that an extract obtained from Japanese cabbage is used as an active ingredient. Further, the anti-stress agent according to the second invention of the present application is characterized in that an extract obtained by extracting from a golden needled vegetable is used as an active ingredient.

[0007] It has been conventionally recognized that Kinzai has a medicinal component for insomnia, anemia, coldness, stiff shoulders, eye strain, hepatic dysfunction, swelling, etc. Effect) and anti-stress effect were not known. As described later, the inventors of the present application conducted an experiment using a golden sardine extract and found that golden sardine has these effects.

[0008] It is preferable that the extract of the golden seaweed contained in the substances according to the first and second aspects of the present invention is extracted using hot water, and the extraction time is preferably 2 to 4 hours. If the extraction time is less than 2 hours, the above effect is not preferably exhibited, and the effect is not changed even if the extraction is performed for 4 hours or more.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION 1
The parts by weight were combined, and the golden soybean extract was extracted with hot water of about 80 ° C. for 2 hours. The following experiment was carried out using the thus-extracted Golden Harvest Extract.

First Example Rats were orally administered 3 g / kg of Golden Harvest Extract for 7 consecutive days, and induced dementia artificially by scopolamine administration. After that, acquisition trial and regeneration trial in passive avoidance learning were performed. Time was measured. As a result, the response latency of the regeneration trial to the acquisition trial was 57.16 for the control group rats.
On the other hand, the response latency of the group to which the Kinzai extract was administered was significantly increased to 174.54 seconds. In addition, the rate of increase in the latency of the regeneration trial with respect to the acquisition trial was 99.4% in the control group rats, whereas the group administered with the Golden Harvest Extract showed a significantly high value of 1099.9%. This experiment revealed that the Kinnasai extract exhibits a remarkable anti-dementia effect in a scopolamine-induced dementia model in rats.

Experiment 1 First, a preliminary study was conducted by the following method. As a test substance, a substance prepared by dissolving the golden needle vegetables extract extracted by the above method in water for injection was prepared.

As reagents, water for injection (Otsuka Pharmaceutical Factory, lot number: K8C73), scopolamine (Sig)
ma, Lot No. 16H0459) and physiological saline (Lot No .: 8C99P, manufactured by Otsuka Pharmaceutical Co., Ltd.).

As experimental animals, 6-week-old Slc: Wi are used.
Star male rats were used. The rats were allowed to freely ingest solid samples (Lab MR Stock, manufactured by Nippon Nosan Kogyo Co., Ltd.) and tap water, and the temperature was 22 ± 2 ° C. and the humidity was 55 ± 1.
5%, lighting time from 8:00 to 20:00, and the number of evocations 1
Preliminary breeding was carried out for one week including the quarantine period in an environment of 3 to 17 times / hour, and rats in which no abnormality was observed in the general condition were used for the test.

Test Method Seven-week-old rats were used based on the following group composition table 1. Group composition table 1

[Table 1]

That is, 3 g of the above test substance was added to 5 rats.
/ Kg / day was orally administered once a day for 7 consecutive days to obtain test rats. On the other hand, water for injection was similarly administered to five control rats. On the seventh day, one hour after administration of the test substance and water for injection, 0.5 mg / kg of scopolamine was subcutaneously administered to both groups of rats, and a memory acquisition trial was performed after one hour.

The acquisition trial was performed using a shuttle direction avoidance experiment box. One side of the experimental box was a bright room, and the other side was a dark room. In addition, only the dark room can be energized. The rats were placed in the light room of the experiment box thus prepared, and the time (latency) until the rats were moved to the dark room was measured for a maximum of 5 minutes. When the rats moved to the dark room, they were shut down and the dark room was energized to give a shock (trial acquisition). The next day, the rats were placed in the light room again and the latency until they moved to the dark room was measured (reproduction trial). According to the following equation, the latency increase rate was calculated for each rat group. Latency increase rate (%) = Reproduction trial latency (second) −Acquisition trial latency (second) / Acquisition trial latency (second) × 100

Statistical processing The average value and standard error of the numerical values obtained for each group were calculated. The significant difference between the groups was determined by performing a variance test using the F test, and in the case of equal variances, using the Student-t test. A risk rate of less than 5% was considered significant.

Test Results The latencies of the acquisition trial and the reproduction trial are shown in FIGS. 1 and 2.
The acquisition trial was almost the same as the control group rat, 39.65 seconds, and the golden needle vegetable extract-administered rat group, 34.14 seconds. Regarding the regeneration trial, the control group rat was 72.74 seconds, and the latency of the regeneration trial was only slightly prolonged with respect to the acquisition trial. The latency was 183.07 seconds, and the latency was significantly prolonged as compared to the control rats. The rate of increase in the latency of the rats of the group receiving the Golden Harvest Extract relative to the latency of the rats of the control group is 151.7%.

FIG. 2 and FIG. 3 show the results of the latency increase rate of the reproduction trial with respect to the acquisition trial. As is clear from FIGS. 2 and 3, the latency increase rate was 410.4% in the control group rats.
In contrast, 81
The rate was 0.2%, and the rate of increase of the rats in the group administered with the Golden Needle Extract was 97.4% higher than that in the control group.

From the results of the preliminary examination described above, although no significant difference was obtained in the latency of the acquisition trial from the control group rats, both the regeneration trial latency and the latency increase rate were higher than the control group rats. Values were observed. Therefore,
In examining the effect of the Golden Harvest Extract on scopolamine-induced dementia rats, the dose of 3 g / kg / day, the administration period once a day for 7 consecutive days, and the oral administration (administration method) evaluate the anti-dementia effect. Was considered reasonable.

[0021] Scopolamine is a drug having an antimuscarinic action, antagonizing acetylcholine at the muscarinic receptor of acetylcholine, and a substance having a memory inhibitory action due to a central action.

[0022] Scopolamine-induced behavioral abnormalities of memory impairment are currently widely used as animal models for amnesia.
It has been reported that scopolamine reduces the performance of memory tasks when administered to humans, and when the postmortem brain of patients with Alzheimer's dementia is analyzed, there are many reports that cholinergic nerve dysfunction is seen. The reason is that the administration of scopolamine does not cause the animals to be in an excessively sedated state, which is convenient for observing behavioral changes. A dementia model using scopolamine is widely used for evaluating anti-dementia drugs, and it is appropriate to use scopolamine as an agent that induces amnesia, that is, memory impairment in the experiments of the present invention.

The above-described experiment method using passive avoidance learning is widely used as an experiment for learning and memory of animals. As described above, passive avoidance learning includes two stages: an acquisition trial and a reproduction trial. This experiment uses a modification in which a mouse or rat prefers a dark room to a bright room, and includes a step-through type in which an experimental box is divided into two rooms and a step-down type in which a platform is provided in one corner of the experimental box. There is.
In both cases, when moving from a certain section to another section, an electric shock is applied, and learning to stay in the original section is performed.

In the experiment of the present invention, a step-through type experiment box was used.
An electric shock was applied from the floor when the person moved into the dark room. Train animals to learn to stay in a light room, with punishment for entering a dark room. A reproduction trial is performed after a lapse of a certain period from the acquisition trial. In the reproduction trial, as in the acquisition trial, the animal is placed in a bright room, and the time from when the animal is placed in the bright room to through movement is measured as a reaction latency.

When accurate learning and memory for the electric shock experience are obtained in the acquisition trial, the response latency at the time of the reproduction trial exceeds the reference response latency (300 seconds). If there is a disability, the response latency will be reduced. Thus, passive avoidance learning is an appropriate index for evaluating the learning and memory state of an animal.

This test Based on the results of the above preliminary study, this test was carried out in accordance with the following group constitution table 2. The test method and statistical processing were performed in the same manner as in the preliminary study. Group composition table 2

[Table 2]

Test Results The latency of the acquisition trial and the regeneration trial of the control group rat and the group of rats receiving the Golden Harvest Extract are shown in FIG. 4 and FIG. The latency of the acquisition trial was 34.19 seconds for the control group rats,
Rats in the group that received the Kinzai extract had a slight shortening of 19.54 seconds.
This shortening is not significant. On the other hand, the latency of the regeneration trial was 57.16 seconds in the control group rats, whereas the rat group with the Golden Harvest Extract administration group was significantly extended to 174.54 seconds. Latency of the rats in the group of the Golden Harvest Extract administered to the control group rats was increased by 205.4%.

FIGS. 5 and 6 show average values of the latency increase rate of the reproduction trial with respect to the acquisition trial in this test. As is clear from these figures and tables, the latency increase rate was 99.9% in the control group rats, and a significantly higher value of 1099.9% in the rats with the Golden Harvest Extract administration group. The rate of increase of the latency increase rate of the rats in the group receiving the Golden Needle Extract relative to the rate of latency increase in the control group rats is 1006.5%.

From the above experimental results, it has been clarified that Kinjasai extract has a remarkable anti-dementia effect on a scopolamine-induced rat model of dementia.

Second Example After a predetermined amount of an aqueous solution of golden soybean extract was orally administered to rats, the rats were immersed in water up to the xiphoid process. After a predetermined period of time, the rate of ulcer formation in the stomach was measured. As a result, the ulcer inhibition rate was 88.8% for the rats to which the golden needle vegetables were administered
High figure was seen

Experiment 2 In order to evaluate the physiological activity of Kinzai, the present inventors examined the anti-stress effect of a rat using a water immersion-restrained stress ulcer model.

Preliminary Experiment Male Wistar / ST rats weighing around 250 g were fasted for 24 hours. However, during this period, free water was supplied. The rats were divided into six groups, and a control group was provided with water, and a test group was provided with an aqueous solution of a golden needle vegetables extract at 500 mg / k.
g, and 5 mL / kg was orally administered to each rat using a sonde. One hour later, each rat was placed in a medicinal stress cage of the University of Tokyo, and immersed in water at a water temperature of 22 ° C. up to the xiphoid process.

After 1 hour, 2 hours and 3 hours after the start of the water immersion, the cage was lifted, and the rats were killed with ether. The abdomen was incised, the pyloric part of the stomach was ligated, 10 mL of physiological saline was injected, and after the ligation of the cardia, the stomach was excised and the serosal side was immersed in 1% formalin solution to form on the inner wall. Gastric ulcer was fixed. After a lapse of 10 minutes, the stomach was incised along the greater curvature, and the contents were wiped to obtain a sample. The formed ulcer was measured using a 10 × stereo microscope, and the area was defined as an ulcer index.

The results of the preliminary experiment are shown in Table 3 below.

[Table 3]

FIGS. 7A and 7B are photographs showing each state of the sample 3 hours after the start of water immersion, wherein FIG. 7A shows the state of the control rat group, and FIG. .

Discussion In this preliminary experiment, it was considered that the golden soybean extract significantly suppressed the formation of ulcers due to water immersion restraint stress. The ulcer inhibition rate of the golden soybean extract was determined from the ulcer coefficient three hours after the start of water immersion, and a high inhibition rate of 88.8% was found.

Volume test The results of the above preliminary test, ie, ulcer formation rate (ulcer index),
It is considered that the effectiveness of the Golden Harvest Extract can be determined about 4 hours after administration of the Golden Harvest Extract from the absorption distribution state of the Golden Harvest Extract. Therefore, in this test, 4 hours was set as the test time.

Water Immersion Restraint Stress Loading Test (Main Test) Male Wistar / ST rats weighing around 250 g were fasted for 24 hours. During this time, water was freely supplied. The rats were divided into 4 groups of 4 rats each, water was added to the control group, 500 mg / kg, 300 mg / kg, 10 mg to the test group.
5 mL of the aqueous solution of the golden needle vegetables adjusted to 0 mg / kg
/ Kg was administered by gavage to rats in each test group using a sonde. After a lapse of 10 minutes, the rats were placed in a straight cage of the University of Tokyo and immersed in water at a water temperature of 22 ° C. up to the xiphoid process.

Four hours after the start of the water immersion, the cage was lifted, and the rats were killed with ether. Cut the abdomen,
The pyloric part of the stomach was ligated, and 10 mL of physiological saline was injected.
After ligation of the cardia, the stomach was removed and the stomach side was immersed in 1% formalin solution to fix the ulcer formed on the inner wall. After a lapse of 10 minutes, the stomach was incised along the greater curvature, and the contents were wiped to obtain a sample. The formed ulcer was measured using a 10 × stereo microscope, and the area was defined as an ulcer index. From the obtained numerical values, the ulcer inhibition rate was determined based on the following equation. Ulcer inhibition rate (%) = control-test group / control x 100

The results are shown in Table 4 below.

[Table 4] 8 to 11 are photographs showing the state of the sample 4 hours after the start of the water immersion, FIG. 8 shows the state of the control group rats, and FIG. 9 shows the state of the rats receiving the 100 mg / kg Golden Harvest Extract group. FIG. 10 shows the state of the rats to which the 300 mg / kg Golden Needle Extract was administered, and FIG.
1 shows the state of rats in the group to which the Japanese radish extract was administered.

[0041] As shown in the above table, the Golden Harvest Extract
It was found that stress ulcers were suppressed depending on the concentration. At a dose of 100 mg / kg, the inhibition rate of about 50% was shown, indicating that the IC ₅₀ (50
% Inhibition) is estimated to be around this.

[0042] In addition, the administration of the Japanese radish extract immediately before the application of stress exerted an anti-ulcer effect, indicating that the Japanese radish extract was immediately effective.

Third Example A plurality of students took Golden Harvest Extract, and a test having the same degree of difficulty was performed before and after a month or two had passed since the administration.
A comparison of the results showed a significant improvement in the points scored after taking.

Experiment 3 (Clinical test) Target: 12 healthy students

Drugs used and administration method For the test students, 600 g of hot water extracted extract of 1 g of Naka Kinzai
1 hour before bedtime every day for 30 days
9 g per dose was taken with hot water. On the other hand, the control students did not take the Golden Harvest Extract for 30 days and were allowed to live a normal life. The medicines that have been conventionally taken were taken as they were. However, taking additional new drugs was prohibited.

Judgment Method The test 1 of the operation was performed before taking the golden needle vegetables extract, and scored. One or two months after the start of the administration, a test 2 having the same difficulty as the test 1 was performed, and the degree of improvement in the score was examined. As a control experiment, the degree of improvement in the score was examined without taking the Golden Harvest Extract. Tables 5 and 6 show the results.
Shown in

[0047]

[Table 5]

[0048]

[Table 6]

Results As is evident from Tables 5 and 6, for the students who did not take the Golden Harvest Extract, there was no significant difference between the scores of Test 1 and Test 2, whereas the students who took Golden Harvest Extract did not. For those who did, there was a clear improvement in post-dose test results.

Table 7 shows the results of the analysis of the properties of the golden seaweed extract used in the present invention.

[Table 7] Note * Nitrogen / protein conversion coefficient: 6.25 ** Calculation formula: 100- (water + protein + lipid + fiber + ash)

[0051]

As described above in detail, the substance containing the Kinzai greens extract according to the present invention as an active ingredient has an anti-dementia effect and an anti-stress effect.

[Brief description of the drawings]

FIG. 1 is a graph showing the results of a preliminary examination test of a first embodiment of the present invention.

FIG. 2 is a table showing the results of a preliminary examination test of the first embodiment of the present application.

FIG. 3 is a graph showing the results of a preliminary examination test of the first embodiment of the present application.

FIG. 4 is a graph showing the results of the main test of the first embodiment of the present invention.

FIG. 5 is a table showing the results of this test of the first example of the present application.

FIG. 6 is a graph showing the results of the main test of the first embodiment of the present invention.

FIG. 7 is a photograph showing a result of a preliminary test of the second embodiment of the present invention.

FIG. 8 is a photograph showing the result of the main test of the second embodiment of the present invention.

FIG. 9 is a photograph showing a result of the main test of the second embodiment of the present invention.

FIG. 10 is a photograph showing the result of the main test of the second embodiment of the present invention.

FIG. 11 is a photograph showing the result of the main test of the second embodiment of the present invention.

Claims (4)

[Claims] 1. An anti-dementia substance comprising, as an active ingredient, an extract obtained by extracting Japanese cabbage. 2. An anti-stress agent comprising an extract obtained by extracting from Japanese cabbage as an active ingredient. 3. The anti-dementia substance according to claim 1, wherein the extract is extracted using boiling water, and the extract is obtained for an extraction time of 2 hours to 4 hours. 4. The anti-stress agent according to claim 2, wherein the extract is extracted using hot water, and the extraction time is 2 to 4 hours.