TWI721407B - Composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes - Google Patents

Abstract

The purpose of the present invention is to propose a composition that contains earthworms as an effective ingredient to inhibit fatty liver and kidney hypertrophy, which is highly likely to be caused by diabetes. The present invention relates to fermented earthworms obtained by mixing the fermented earthworms in the fermentation broth obtained by inoculating yeast in the vegetable and fruit extracts and fermenting the earthworms, and then mixing the fermented earthworms with panax notoginseng to ferment and dry them to obtain a fermented mixture. After mixing the fermented mixture with ants and ginkgo biloba leaves, a composition that inhibits fatty liver and kidney hypertrophy caused by diabetes is obtained.

Description

Composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes

The present invention relates to a composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes and its manufacturing method.

In East Asia, earthworms have been used as antipyretic analgesics and other crude drugs since ancient times. In addition, in recent years, diabetes treatment agents (Patent Document 1) and anti-hyperlipidemia agents (Patent Document 2) that use earthworm components as active ingredients have also been proposed. ), blood pressure regulator (Patent Document 3), etc.

[Prior Technical Document] [Patent Document]

[Patent Document 1] Japanese Patent Publication No. 07-080778

[Patent Document 2] Japanese Patent Publication No. 07-080777

[Patent Document 3] Japanese Patent Publication No. 07-039349

As mentioned above, considering the various functions of earthworms, this application group After further research on the effectiveness of this earthworm, the team found that the earthworm component has a novel effect of inhibiting the onset of fatty liver and kidney hypertrophy caused by diabetes.

Therefore, the purpose of the present invention is to propose a composition that contains earthworms as an active ingredient to inhibit fatty liver and kidney hypertrophy which is highly likely to be caused by diabetes, and its manufacturing method.

The gist of the present invention will be explained with reference to the attached drawings.

The present invention relates to a composition characterized in that it contains earthworms as an effective ingredient and can inhibit fatty liver and kidney hypertrophy caused by diabetes.

In addition, the present invention relates to the composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes as claimed in claim 1, wherein the earthworm is a fermented earthworm obtained by fermenting a fermentation broth obtained by inoculating yeast in a vegetable and fruit extract.

The present invention relates to the composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes as claimed in claim 1, wherein the earthworm is Eisenia vulgaris.

The present invention relates to a composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes as claimed in claim 2, wherein the earthworm is Eisenia vulgaris.

The present invention relates to a composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes as claimed in any one of claims 1 to 4, which contains at least one of Panax notoginseng, ants, and ginkgo biloba.

The present invention relates to a composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes as claimed in any one of claims 1 to 4, which contains notoginseng, ants, and Ginkgo biloba.

The present invention relates to the composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes as claimed in claim 5, wherein the ant is a fermented ant that has been fermented.

The present invention relates to the composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes as claimed in Claim 6, wherein the ant is a fermented ant that has been fermented.

The present invention relates to the composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes as claimed in claim 5, wherein the ginkgo biloba is a fermented ginkgo biloba extract obtained by fermenting the extract from the ginkgo biloba extract.

The present invention relates to a composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes as claimed in claim 6, wherein the ginkgo biloba is a fermented ginkgo biloba extract obtained by fermenting an extract from the ginkgo biloba.

The present invention relates to a composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes as claimed in claim 7, wherein the ginkgo biloba is a fermented ginkgo biloba extract obtained by fermenting the extract from the ginkgo biloba extract.

The present invention relates to the composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes according to claim 8, wherein the ginkgo biloba is a fermented ginkgo biloba extract obtained by fermenting an extract from the ginkgo biloba.

The present invention relates to a method for producing a composition that inhibits fatty liver and kidney hypertrophy caused by diabetes. The fermented earthworm is obtained by mixing earthworms in the fermentation broth obtained after inoculating yeast in vegetable and fruit extracts and fermenting the earthworms. , And then mix the fermented earthworms and panax notoginseng to ferment and dry to obtain a fermented mixture, and mix the fermented mixture with ants and ginkgo leaves to obtain It can inhibit fatty liver and kidney hypertrophy caused by diabetes.

Since the constitution of the present invention is as described above, it is possible to suppress the onset of fatty liver and kidney hypertrophy, which are highly likely to be caused by diabetes, by ingesting the present invention.

[Figure 1] In Test 1 of this example, the measurement results of the weight of each organ such as the liver, kidney, and heart, as well as the weight of fat around the kidney, and the fat around the testicle.

[Figure 2] The weight of the liver per 100 g of body weight in Test 1 of this example.

[Figure 3] An enlarged view of the kidney weight in Figure 1.

[Figure 4] In Test 1 of this example, the ratio of total lipids in the liver.

[Figure 5] The weight of total cholesterol contained in the liver in Test 1 of this example.

[Figure 6] The measurement results of urine albumin in Test 1 of this example.

[Figure 7] In Test 1 of this example, the measurement result of plasma fibrogen activator inhibitor type I (PAI-1).

[Fig. 8] In Test 1 of this example, the measurement result of euglobulin clot lysis time (ECLT).

[Figure 9] In Test 1 of this example, it shows the evolution of blood glucose level.

[Figure 10] The measurement result of glycosylated hemoglobin (HbA1c) in Test 1 of this example.

[Figure 11] The results of insulin measurement in Test 1 of this example.

[Figure 12] The measurement result of amylase in Test 1 of this example.

[Figure 13] The measurement result of uric acid in Test 1 of this example.

[Figure 14] In Test 1 of this example, the measurement result of glutamate oxalate transaminase (GOT).

[Figure 15] The measurement result of alkaline phosphatase (ALP) in Test 1 of this example.

[Figure 16] The measurement result of lactate dehydrogenase (LD) in Test 1 of this example.

[Fig. 17] In Experiment 1 of this example, the tissue image after observation of the kidney section tissue.

[Figure 18] From the tissue image in Figure 17, a graph showing the proportion of the interstitial matrix in the glomerulus quantified.

[Fig. 19] The tissue image after observation of the aortic arch tissue in Test 1 of this example.

[Fig. 20] From the tissue image in Fig. 19, a graph showing the thickness of the vessel wall of the aortic arch digitized.

[Fig. 21] A graph of body weight changes in Test 2 of this example.

[Fig. 22] The measurement result of glycosylated hemoglobin (HbA1c) in Test 2 of this example.

[Figure 23] The results of insulin measurement in Test 2 of this example.

[Figure 24] The results of the measurement of adiponectin in Test 2 of this example.

[Figure 25] In Test 2 of this example, the measurement result of glutamate oxalate transaminase (GOT).

[Figure 26] The measurement result of alkaline phosphatase (ALP) in Test 2 of this example.

[Figure 27] The measurement result of leucine amino peptidase (LAP) in test 2 of this example.

[Figure 28] Measurement results of total blood cholesterol in Test 2 of this example.

[Figure 29] The measurement result of blood urea nitrogen in Test 2 of this example.

[Figure 30] In Test 2 of this example, the measurement result of creatinine clearance rate (Ccr).

[Figure 31] The measurement result of urine albumin in Test 2 of this example.

[Figure 32] Results of measurement of liver weight in Test 2 of this example.

[Figure 33] In Test 2 of this example, the ratio of total lipids in the liver.

[Figure 34] In Test 2 of this example, the ratio of triglycerides in the liver.

[Figure 35] The weight of total cholesterol in the liver in Test 2 of this example.

[Figure 36] In Test 2 of this example, when the euglobulin clot is dissolved The measurement result of the time (ECLT).

[Fig. 37] In Test 2 of this example, the measurement result of plasma fibrogen activator inhibitor type I (PAI-1).

[Figure 38] In Experiment 2 of this example, it shows the proportion of the interstitial matrix in the glomerulus.

The effect of the present invention is revealed and briefly described with the embodiments of the present invention considered to be more suitable.

The composition of the present invention for inhibiting fatty liver and kidney hypertrophy caused by diabetes can improve the lipid metabolism function decreased due to diabetes by, for example, orally ingestion, and can also inhibit the reduction of lipid metabolism function caused by diabetes.

Furthermore, it can improve the small blood vessel disorder caused by diabetes, and can inhibit the onset of small blood vessel disorder caused by diabetes (confirmed by the ZDF rat (Zucker Diabetic Fatty Rat) using type II diabetes model rats The test result is known).

Therefore, the onset or progression of fatty liver and kidney hypertrophy caused by diabetes can be suppressed.

[Example]

The specific embodiments of the present invention are described on the basis of the drawings.

The present invention relates to a composition that uses earthworms as an effective ingredient to inhibit fatty liver and kidney hypertrophy that are prone to complications when type II diabetes occurs. Specifically, it is composed of earthworms, fermentation broth, Panax notoginseng (also known as Tianqi, scientific name: Panax motoginseng Berk), ants, and ginkgo leaves.

More specifically, the composition for inhibiting fatty liver and kidney hypertrophy caused by diabetes (hereinafter referred to as the composition) of this embodiment is mixed with earthworms in the fermentation broth obtained by inoculating yeast in the vegetable and fruit extracts. Fermenting the earthworms to obtain fermented earthworms, and then mixing the fermented earthworms with Panax notoginseng to ferment and dry to obtain a fermentation mixture, which can be obtained by mixing the fermentation mixture with ants and ginkgo leaves.

Hereinafter, this composition will be described in detail.

The earthworm used in this example is Eisenia fetida (scientific name: Eisenia fetida).

In addition, the fermentation broth is a vegetable and fruit extract extracted from vegetables and fruits, which is inoculated with yeast (in this example, multiple types of yeasts, such as Nihwa and Candida) are used to ferment, Specifically, 5kg of mung bean sprouts, 1.8kg of cabbage, 0.5kg of cabbage, 0.5kg of cabbage, 0.5kg of small pine cabbage, 0.6kg of lotus root, 0.5kg of basil, 10kg of potato, 2kg of cucumber, 0.6kg of burdock, 45kg of papaya, Cut 15kg of pineapples, 2kg of bananas, 5kg of apples, and 1kg of lemons into appropriate sizes. Then add 81kg of anhydrous dextrose, 9kg of water-containing dextrose, and 0.21 yeast. The juicer squeezes the fermented fermented product, and filters the squeezed fermented liquid (filter screen is 120 mesh).

Moreover, the vegetables and fruits used to obtain the vegetable and fruit extract are not restricted to the above-mentioned vegetables and fruits, and vegetables and fruits other than the above may also be used.

In addition, Panax notoginseng is processed in powder form.

Ants are two kinds of ants, namely, Polyrhachisviva Roger (scientific name: Polyrhachisviva Roger) and Black Mountain ants (scientific name: Formica japonica). Specifically, these ants are ants that have been fermented.

More specifically, the fermented ant system is mixed with papaya powder, made into a powdered black spiny ant (hereinafter referred to as black spiny ant powder), and made into a powdered black mountain ant (hereinafter referred to as black mountain ant powder) And water, and then inoculate yeast (raw yeast) into it and ferment it, and then dry the fermented product and make it into powder.

A more detailed description is to mix 38.2kg of papaya powder, 1.82kg of black spiny ant powder, 7.28kg of black mountain ant powder and (40℃) 441 water in a working room set at a room temperature of 28℃, and then inoculate 7.7 g yeast (raw yeast), leave it to stand for 24 hours to ferment, and then dry the fermented person at 25°C for 72 hours, and then dry the water to below 10% at 21°C, and dry it into water If the content is less than 10%, after coarsely pulverizing with a 2mm sieve, then pulverizing with a 0.2mm sieve to make a powder.

In addition, the aforementioned papaya powder is produced by shredding the papaya into shreds or pulverizing it with a pulverizer, adding sugars to extract the papaya enzyme, adding yeast to the papaya extract to make the yeast adhere to the original The wild yeast on the papaya is fermented and proliferated, and then lactic acid bacteria are added to distinguish the liquid component obtained after symbiotic culture of the aforementioned yeast, wild yeast and lactic acid bacteria, and yeast and lactic acid bacteria are further added to the liquid component to ferment and mature It is obtained by post-drying (refer to Patent No. 3370302 of Kondo et al.).

Ginkgo biloba is a ginkgo biloba extract extracted from ginkgo biloba leaves. Specifically, it is a fermented ginkgo biloba extract obtained by fermenting the ginkgo biloba extract.

More specifically, the fermented ginkgo biloba extract is obtained by cooking the ginkgo biloba leaves, then heating and concentrating the extracted extract, mixing the concentrated extract with anhydrous glucose and fermented calcium, and then inoculating yeast to ferment it , And finally the fermented product is dried and made into powder.

A more detailed description is that 50kg of ginkgo biloba and 5001 water are placed in a kneading machine and heated and boiled for 1 hour to extract the extract. The extracted extract is filtered with #350 and #500 filters, and the process is heated. The filtered extract was concentrated to 601, and when the liquid temperature of the concentrated extract was still high, it was mixed with 17.5kg of anhydrous glucose, and then the liquid temperature was lowered to 30℃ and mixed with 5kg of fermented calcium. After that, the yeast was inoculated, and the room temperature was set to Aerobic fermentation is carried out for 24 hours in a working room at 28°C. After fermentation, the room temperature is maintained at 23~24°C to dry it. During drying, 5kg of brown rice and 22.5kg of starch are made into powder form (in this example, the manual is used). Powder starch) is divided into two days and mixed in half each. After the completion of the mixing operation, the room temperature is reduced to 20°C from the next day, and the moisture content is reduced to less than 5% after the completion of the mixing operation. The dried product is crushed with a 0.2mm screen and made into powder.

The aforementioned fermented calcium is in the aforementioned fermentation broth. The fermentation broth with a fermentation period of 2 weeks is used. Compared with the fermentation broth of 11, calcium powder is added in a ratio of 0.3kg (from 70wt% Hokuriku shell fossil, 28wt% shell calcium, 2wt% Fish calcium composition), stir for about 2 weeks, and then use the ratio of 5.5ml kelp extract relative to the fermentation broth of 11, and add 0.21 water to the fermentation broth of 11 Example (In this example, a mixture of kelp extract and water was added to the fermentation broth), the next day after adding the kelp extract and water, the room temperature was raised to 25°C, and the next day, the room was again The temperature is lowered to 21°C, and the moisture content is reduced to less than 8% by drying, and the dried product is coarsely pulverized with a 2mm sieve, and then pulverized with a 0.2mm sieve to make a powder.

Next, the manufacturing method of this composition will be described in detail. And in the following description, 1kg Eisenia vulgaris is used (100g after drying, the blending ratio is 15wt%).

First, remove the sand and mud from the earthworm Eisenia vulgaris, clean it, and then smash it into granules of less than 2mm.

Specifically, the sand and mud in the earthworms are removed in a mixer, and the washed earthworms are washed with water (3.51 water is added to the weight of 1kg of earthworms), and then crushed by the mixer 40 ~80 minutes, that is, you will get the mud-like earthworm Aisheng. In addition, the liquid temperature will rise during the pulverization process, because when the rising liquid temperature exceeds 30°C, the effective components of Eisenia vulgaris may be deteriorated or destroyed. During the pulverization process, the temperature should not exceed 30°C. The temperature of the water added should be lower than 30°C. Below 15°C is preferred.

Secondly, the crushed earthworm Aisenia vulgaris was mixed with 215ml of fermentation broth (20wt% blending ratio), 125ml of lactic acid bacteria, and left to stand at 25°C for 17-21 hours to obtain the fermented Aisei. earthworm. In addition, the lactic acid bacteria in this embodiment use K-1 bacteria or Candida albicans with good antibacterial effect. If the lactic acid bacteria used can inhibit the proliferation of coliforms and coliforms, they can be cured during fermentation and drying. There are no special restrictions on the killer.

Secondly, the fermented Eisenia vulgaris and 180 g of Panax notoginseng powder (the blending ratio is 27 wt%) are mixed, and the mixed first mixture is fermented and dried to the extent of no moisture.

Specifically, the room temperature is kept at 23°C, while blowing air into the first mixture, it is fermented and dried for 48 to 72 hours. (In the stage of fermentation and drying, the action of the lactic acid bacteria mixed during the production of fermented earthworms is used to prevent coliform bacteria, Coliform bacteria), and after the fermentation and drying are completed, keep the room temperature at 20-28°C and humidity below 18% for about 2 weeks, and perform air drying to dry the water content of the aforementioned first mixture to below 5% .

Then, in the dried first mixture, 20 g of fermented ants (blending ratio: 3 wt%) and 233 g of fermented ginkgo biloba extract (blending ratio: 35 wt%) were mixed to obtain a second mixture.

Finally, the second mixture was coarsely pulverized with a 2mm mesh screen and dried, and then crushed with a 0.2mm mesh screen to form a powder, and the powder was filled into capsules to complete the production. And the final form is not limited to the capsule form, it can be maintained in a powder form or made into a tablet form.

In addition, in this embodiment, the ingredients (materials) can be blended according to the aforementioned blending ratios, and the fermented Eisenia vulgaris is 12-18wt%, the panax notoginseng powder is 22-32wt%, and the fermented ants is 2~4wt%, the fermented ginkgo leaf extract is 28~42wt%, and the fermentation broth is in the range of 16~24wt%.

Next, the effect of this composition will be explained.

In this example, a second test for confirming the effect of test 1 and test 2 was performed. Test 1 is simply to confirm the effect of the composition, test 2 In the middle system, the effects of the earthworm (earthworm) components that were not confirmed in Test Example 1, and the effects of other components and the effects on early symptoms were confirmed. First, test 1 will be described in detail.

<<Experiment 1>>

In this test, ZDF rats (Zucker Diabetic Fatty Rats), which are type II diabetes model rats, were used to compare ZDF rats administered with general diets that did not contain the above-mentioned composition with ZDF administered with diets containing the composition. The difference in the state of the rat confirms the effect of this composition.

Specifically, 20 7-week-old male ZDF rats were prepared, and these rats were divided into 2 groups with almost the same average weight and blood glucose level, and a group of 10 rats. One group was given a general feed that did not contain this composition. (Control group feed) Control group (hereinafter referred to as C group), and the other group is Eisenia fetida group (hereinafter referred to as EF group) given the feed containing this composition.

The general feed given to the C group is based on AIN-93G, the adjusted feed using casein as the protein source. In addition, the feed given to the EF group will be given 5% (by weight) of the corn starch contained in the general feed of the C group. ) Feed replaced by this composition.

In addition, the feeding method is to give each group each feed for 12 weeks (7 weeks old to 19 weeks old), and feed the rats by feeding. And you can take in water freely. In addition, because the fasting blood glucose is measured every week, fasting 18 hours before the measurement.

For each ZDF of the C group and EF group bred according to the above conditions Rats, measure blood glucose level, glycosylated hemoglobin (hereinafter referred to as HbA1c), insulin, organ weight (kidney, liver, heart, peripheral fat, and peripheral fat), euglobulin clot dissolution time (hereinafter referred to as ECLT) At the same time, observe the kidney slices and the tissues of the aortic arch, compare the differences between the C group and the EF group and confirm the effect.

Figures 1 to 3 are graphs showing the measurement results of the weights of the organs of the kidney, liver, and heart, as well as the weight of fat around the kidney and the fat around the testis.

In the measurement of the equal weight of the organs, as shown in Figure 1, the correlation between the kidney and the liver shows a significant difference (t test, p<0.01) (the correlation between the heart, the fat around the kidney, and the fat around the testicle, group C and There are no significant differences between the EF groups).

In detail, from the liver weight measurement results shown in Figure 2 and the kidney weight measurement results shown in Figure 3, it is clear that there are significant differences between the respective C group and the EF group (t test, p<0.01), Compared with group C and group EF, liver weight and kidney weight showed lower results.

Fig. 4 is a graph showing the weight of total lipids contained in the liver, and Fig. 5 is a graph showing the weight of total cholesterol (T-cho) contained in the liver.

In the determination of the weight of total lipid and total cholesterol contained in the liver, both total lipid and total cholesterol showed significant differences between group C and group EF (t test, p<0.01), compared to group C by EF Group, total lipid and total cholesterol all showed lower results. In addition, in rats (healthy rats) that are general reference values, the total liver lipids are about 4% and the total cholesterol is about 0.4% relative to the total weight of the liver.

From the liver weight measurement results shown in Figure 2 and the total liver weight shown in Figure 4 The lipid weight measurement results and the measurement results of liver total cholesterol weight shown in Figure 5, the ratio of total lipid and total cholesterol relative to liver weight of C group and EF group was calculated, and group C was compared with normal rats , The ratio of total lipid and total cholesterol relative to liver weight both showed high values, but the ratio of total lipid and total cholesterol relative to liver weight of EF group was the same value as that of normal rats.

From this result, the liver weight of EF group ZDF rats is significantly lower than that of C group ZDF rats, which is believed to be caused by the difference in total lipids and total cholesterol of the liver.

In other words, it is believed that by ingesting the composition in ZDF rats of the EF group, the reduction in lipid metabolism caused by diabetes is improved, and the lipid metabolism is close to normal, or the composition is inhibited by the ingestion of the composition. The lipid metabolism function is reduced, and the lipid metabolism is performed as normal, and the fat accumulation in the liver is inhibited. The total lipid and total cholesterol of the liver do not increase, and the liver weight is almost the same as that of the average rat.

Fig. 6 is a graph showing the measurement results of albumin in urine.

In this urine albumin measurement, there was no significant difference between the C group and the EF group from the start of the test (7 weeks of age) to the 6th week (13 weeks of age), but the 8th week (15 weeks of age) after the start of the test ), there is a significant difference between group C and group EF (t test, p<0.05). Compared with group C, the urine albumin of group EF is shown as a low value. In addition, the 10th week after the start of the test (17 After weeks of age), the meaningful difference disappeared. However, from about 4 weeks after the start of the test to the end of the test, compared with group C, the value of urinary albumin in group EF is lower, which shows that there is a tendency to inhibit albumin excretion. to.

Generally speaking, when the course of diabetes progresses, the complications of diabetic nephropathy are often seen. When the diabetic nephropathy develops, it is known that albumin that is hardly present in the urine will leak out, which is effective It was judged to be the initial diabetic nephropathy, and the leakage of albumin in the urine is thought to be caused by the damage to the small blood vessels caused by diabetes.

That is, it is believed that by ingesting this composition, the effect of improving fibrinolytic reaction and improving the damage of small blood vessels by ZDF rats of EF group is lower than that of C group of urine albumin.

In addition, the kidney weight of ZDF rats in the EF group ingesting the composition is significantly lower than that in the ZDF rats in the C group not ingesting the composition. It is believed that this fibrin The increase in the lysis reaction inhibits the onset of small blood vessel damage due to diabetes, does not cause a decrease in kidney function, and then improves or inhibits the symptoms of renal hypertrophy.

Figure 7 is a graph showing the measurement results of plasma fibrogen activator inhibitor type I (hereinafter referred to as PAI-1) in blood.

In the measurement of PAI-1, there is a significant difference between the C group and the EF group (t test, p<0.05). Compared with the C group, the PAI-1 value of the EF group is lower.

The decreased lipid metabolism of PAI-1 due to diabetes and the hypertrophy of fat cells accompanied by the accumulation of visceral fat caused by the decreased lipid metabolism increase its release. When the release of PAI-1 increases, it is known that fiber The proteolytic reaction is reduced and thrombus is easily formed.

That is, from the measurement result of the PAI-1, it can be known that the composition can The improvement of the decrease in metabolism caused by diabetes suggests that it has the effect of inhibiting the decrease of fibrinolytic response caused by diabetes.

Figure 8 is a graph of ECLT measurement results. In addition, in this ECLT measurement, healthy rats without diabetes mellitus other than ZDF rats of group C and EF were used, specifically, SD rats were used, and the ECLT value of the SD rats was used as the reference point. Compare the ECLT value of the SD rats with the ECLT values of the C group and the EF group.

The ECLT is combined with thrombin, which is the blood coagulation factor in the measurement of euglobulin clot dissolution time, to make an artificial thrombus. The fibrinolysis reaction is evaluated by measuring the dissolution time of the clot. The longer the ECLT, the fibrinolysis. The reaction decreases.

In this experiment, as shown in Figure 8, there is a significant difference between group C and group EF (t test, p<0.01), which shows that compared with group C, the ECLT given to the EF group of this composition is a result of a low value. At the same time, it was confirmed that the ECLT value of ZDF rats administered with the EF group of this composition was equivalent to the ECLT value of healthy SD rats.

From the results of the ECLT measurement, it was confirmed that the composition has the effect of improving the reduction of fibrinolytic response due to diabetes, or has the possibility of inhibiting the reduction of fibrinolytic response due to diabetes.

That is, in general, the onset of complication of thrombosis is a major problem when diabetes occurs. However, this composition has been confirmed to have the possibility of inhibiting the onset of thrombosis caused by diabetes.

Fig. 9 is a graph showing the evolution of blood glucose levels during fasting, and Fig. 10 is a graph showing the results of HbA1c measurement.

Generally speaking, HbA1c is more easily formed as the blood sugar level is higher, and it increases significantly as the blood sugar level increases at the onset of diabetes.

In this experiment, there was no significant difference in blood glucose and HbA1c between group C and EF, and both groups showed high blood glucose and HbA1c. In addition, the HbA1c value of the general ZDF rat is about 9.4% at 18 weeks of age (refer to the data of Charles River in Japan). Compared with this test, it shows that the HbA1c value is slightly lower, but the self-blood glucose measurement result has been confirmed to be maintained at a high value. There is no doubt about the HbA1c value based on the blood glucose status.

Figure 11 shows the results of insulin measurement.

ZDF rats may cause excessive insulin secretion in the early stage of diabetes, but it can be known that the secretion ability gradually disappears around 18 weeks of age, and the insulin concentration decreases.

In this test, as shown in Fig. 11, the EF group ingesting the composition showed a slight tendency to have a higher insulin secretion ability, but there was no significant difference between the C group and the EF group.

From the blood glucose level, HbA1c, and insulin measurement results, it can be known that the composition does not directly act on the blood glucose level improvement mechanism.

That is, this composition does not inhibit the onset of fatty liver and kidney hypertrophy caused by diabetes by improving the symptoms of hyperglycemia, but directly acts on the functions of the liver and kidneys, and has the function of improving the reduced lipid metabolism caused by diabetes. Reduce the effect of kidney function, or it is believed to have the effect of inhibiting the reduction of liver lipid metabolism and kidney function caused by diabetes.

In addition, this test focuses on total protein (TP), albumin (Alb), zinc sulfate turbidity test (ZTT), revanol turbidity test (TTT), total bilirubin (T-BIL), glutamate oxalate transaminase (GOT), glutamate pyruvate transaminase (GPT), alkaline phosphatase (ALP), lactate dehydrogenase (LD), glutamine transferase (γ-GT), leucine amino peptidase (LAP), cholinesterase (CHE), creatine kinase (CK), acid phosphatase (ACP), amylase ( AMY), uric acid (UA), urea nitrogen (BUN), creatinine (Cre), serum iron (Fe), total cholesterol (T-cho), neutral fat (TG), HDL cholesterol (HDL), arteriosclerosis index 24 items such as (AI) and LDL cholesterol (LDL) are measured and analyzed by blood tests.

Among the 24 items, the two items of amylase and uric acid have significant differences between group C and group EF (t test, p<0.01).

Amylase-based diabetes and renal insufficiency are items that tend to have high values. However, in this test, as shown in Figure 12, the EF group shows a significant low value. From this result, it can be inferred that this composition has the ability to inhibit diabetes. The onset of diabetic nephropathy or the effect of delaying its progress, or the effect of improving the symptoms of diabetic nephropathy.

In addition, the uric acid value is related to items that exhibit high values such as metabolic syndrome. However, in this test, as shown in Figure 13, the EF group shows a significant low value. From this result, it is considered that the present composition has the ability to metabolize uric acid. The possibility of the impact caused by the system.

In addition, although there are no significant differences, as shown in Figures 14-16, the three items of glutamate oxalate transaminase (GOT), alkaline phosphatase (ALP), and lactate dehydrogenase (LD) , Show the result of good value in EF group. It is believed to imply that this composition has the ability to inhibit liver function and reduce liver function. The possibility of low, or, the possibility of improving the reduced liver function.

Fig. 17 is a tissue image showing the result of tissue observation of a kidney section, and Fig. 18 is a chart of digitizing the tissue image.

When observing the kidney section tissue, as shown in Fig. 17, the EF group of the present composition was ingested, and the suppression of the increase of the annulus interstitial matrix was observed.

In addition, comparing the tissue images obtained by digitizing the observations, the results found that there is a significant difference between the C group and the EF group (t test, p<0.05). Compared with the C group, the EF group has the ring stroma The ratio of the area of the glomerulus occupied by the matrix is small, that is, it shows that the increase of the annulus interstitial matrix is suppressed.

This annulus interstitial matrix is responsible for supporting the glomerulus in the kidney. It is known that the annulus interstitial matrix is enlarged due to high blood sugar and other reasons in diabetes, and it oppresses the capillaries of the surrounding glomeruli. This tendency was also observed in group C of this composition, but ingestion of group EF of this composition inhibited the increase in the annulus interstitial matrix, and no symptoms of compression of the capillaries of the glomeruli were found.

In addition, the capillaries compressed by the annulus interstitial matrix narrow the lumen (blood circulation channels), which makes blood flow difficult. The blood that can be filtered by the glomerulus is reduced due to the poor blood flow. Furthermore, the pores on the capillary tube wall that filter blood take on the responsibility of a filter. The thicker the tube wall, the larger the hole, a large amount of protein will be lost. As a result, the symptoms of proteinuria will appear.

In this test, the EF group can inhibit albuminuria, which is believed to be due to the inhibition of the hypertrophy of the interstitial matrix.

In addition, FIG. 19 is a tissue image showing the result of observing the aortic arch tissue, and FIG. 20 is a diagram of digitizing the wall thickness of the arterial arch vessel from the tissue image.

When observing the aortic arch tissue, as shown in Fig. 19, the EF group of this composition was taken, and it was observed that the hypertrophy of the aortic arch vascular wall showed a significant tendency to inhibit.

Furthermore, comparing the tissue images obtained by digitizing the observations, it was found that compared with group C, group EF has thinner blood vessel wall thickness. Although there is no significant difference, it is confirmed that the value is a significant tendency that can be suppressed Hypertrophy.

Secondly, test 2 is described in detail.

<<Experiment 2>>

The purpose of this test is to confirm the effect of the earthworm (Eisenia vulgaris) in this composition, and the effects of components other than earthworm (Eisenia vulgaris). Furthermore, the test period was set to a short period to confirm the effect on early symptoms. .

Specifically, it was the same as in Test 1, using ZDF rats, comparing ZDF rats that were given a general feed that did not contain the composition, and ZDF rats that were given a feed that contained the composition, and were given the composition containing the composition. However, there is no difference in the state of the ZDF rats in the feed that does not contain earthworms (earthworms), which do not contain earthworms, and confirm that the earthworms in this composition are different from those other than the earthworms. effect. And the aforementioned non-earthworm composition is simply to remove the earthworms in this composition The blending ratio of the earthworm (Eisenia vulgaris) and other components is the same as this composition.

Specifically, the ZDF rat system was the same as in Test 1. The 7-week-old male rats were prepared so that the weight and blood glucose levels of these rats were almost the same on average. The same composition as in Test 1 was divided into two groups that did not contain this composition. The C group of the general feed (control group feed) of the animal, the EF group of the feed containing this composition, and the Non-Eisenia fetida group (hereinafter referred to as the NE group) of the feed containing no earthworm composition.

The feed given to the C group and EF group is the same as in Experiment 1. In addition, the feed given to the NE group will be given 4.25% (weight) of the corn starch contained in the general feed of the C group as a composition that does not contain earthworms Replace the feed.

In addition, the feeding method was carried out under the same conditions as in Test 1, except that the test period was set to 10 weeks (7 weeks to 17 weeks of age).

For the ZDF rats of group C, group EF, and group NE that were bred under the above conditions, the weight change, HbA1c, insulin, adiponectin, liver function markers (in the blood), total blood cholesterol, liver lipids, ECLT, PAI-1, the ratio of interstitial matrix, compare the difference between C group, EF group and NE group, confirm the effect of earthworm (Eisenia vulgaris) in this composition, and the effect of other components Confirm the effect on early symptoms.

Fig. 21 is a graph showing the evolution of body weight. As shown in Figure 21, the C group almost did not gain weight around the 5th week (12 weeks of age), and after the 6th week (13 weeks of age), the EF group and the NE group showed signs of interest in body weight. The difference in meaning (LSD test, p<0.01), there is a meaningful difference in the decline in growth (there is no meaningful difference between the EF group and the NE group).

Figure 22 is a graph showing the measurement results of HbA1c. In Test Example 1, there was no significant difference between the C group and the EF group, but in this experiment, there were significant differences between the C group and the EF group, the C group and the NE group (LSD test, p<0.05), the EF group and the EF group The significant HbA1c value is suppressed between NE groups (there is no significant difference between EF group and NE group). It is believed that this result is due to the observation of HbA1c value in the early stage of diabetes.

In addition, even if the NE group does not contain the earthworm (Eisenia vulgaris) component, the HbA1c value was found to be suppressed. It was confirmed that components other than the earthworm (Eisenia vulgaris) component have the effect of suppressing the HbA1c value.

Figure 23 is a graph showing the results of measuring insulin. As in Experiment 1, there was no significant difference between the C group and the EF group. In addition, between the C group and the NE group, and between the EF group and the NE group, there was also no significant difference between the results.

Figure 24 is a graph showing the results of measuring adiponectin (a novel item not measured in Test Example 1). As shown in Figure 24, there are significant differences between group C, EF group and NE group (LSD test, p<0.05). Between group EF and NE group, significant adiponectin value results (EF group and NE group). There is no meaningful difference between groups).

The adiponectin is known to reduce the amount of fat in diabetes, and is related to insulin resistance, etc., and has the effect of lowering the blood sugar level. In addition, it is generally believed that the reduction of adiponectin will cause arteriosclerosis.

Since the NE group that does not contain earthworms (Eisenia vulgaris) components, the value of adiponectin is also found to be high. It is confirmed that the components of earthworms (Eisenia vulgaris) are The external ingredients have the effect of inhibiting the reduction of adiponectin. In addition, the above-mentioned effect of inhibiting the HbA1c value is considered to be the result of the improvement of insulin resistance by inhibiting the reduction of adiponectin.

Figures 25-27 are the measurement results of liver function markers (in the blood). Specifically, Figure 25 is based on glutamate oxalate transaminase (GOT), Figure 26 is alkaline phosphatase (ALP), and Figure 27 is Leucine aminopeptidase (LAP) measurement results. Figure 28 shows the measurement results of total cholesterol in blood.

Leucine aminopeptidase, one of the liver function markers, is known to increase in liver disease. In addition, it is also known to be an item that also increases in fatty liver. In addition, it is known that total cholesterol in the blood can be caused by diabetes. Progress is also an item that will increase.

There was no significant difference in each item in Experiment 1 (Glutamate oxalate transaminase and alkaline phosphatase found significant tendency), but in this experiment, as shown in Figures 25-28, each item showed C Significant differences between the EF group and the NE group (LSD test, p<0.05).

That is to say, in this test, the EF group and the NE group significantly inhibited the increase of glutamine oxalate transaminase, alkaline phosphatase, and leucine aminopeptidase, which are markers of liver function, and also significantly inhibited the blood The increase in total cholesterol, the EF group and the NE group meaningfully inhibit the decrease of liver function, suggesting the result of improving lipid metabolism (there is no significant difference between the EF group and the NE group).

Even if the NE group does not contain the earthworm (Eisenia vulgaris) component, there is a significant difference in each item compared with the C group. It is confirmed that the components other than the component of the earthworm (Eisenia vulgaris) have the effect of inhibiting the reduction of liver function, and Improve the effect of lipid metabolism.

Figures 29 and 30 are the measurement results of renal function markers. Specifically, Figure 29 is the blood urea nitrogen (BUN), and Figure 30 is the measurement results of the creatinine clearance rate (Ccr).

The blood urea nitrogen and creatinine clearance rate of this renal function marker are jointly used as the items of renal function indicators in diabetes. It is known that blood urea nitrogen will increase when the kidney filtration function decreases. In addition, due to the creatinine clearance rate It represents the filtering capacity of the kidneys, and is an item that is known to decrease during the process of diabetic nephropathy.

In Experiment 1, there was no significant difference in the measurement results of blood urea nitrogen. In this experiment, there were significant differences between the C group, the EF group, and the NE group (LSD test, p<0.05). Although there is no significant difference in the creatinine clearance rate, there is a significant tendency between the C group and the EF group and the NE group.

That is, the EF group and NE group in this test significantly inhibit the increase of blood urea nitrogen as a marker of renal function, and at the same time have a tendency to inhibit the decrease of creatinine clearance rate, suggesting that EF group and NE group have the effect of inhibiting the decrease of renal function (There is no significant difference between the EF group and the NE group).

Figure 31 The results of measuring albumin in urine.

In test 1, a significant difference was found 8 weeks after the start of the test, but in this test, from the 6th week (13 weeks of age) from the start of the test, there were significant differences between the C group, the EF group, and the NE group (LSD test, p<0.05), and finally (10 weeks after the start of the test), there were significant differences between the groups (LSD test, p<0.05).

Specifically, compared with group C, group EF can significantly inhibit urine The amount of albumin leakage, further, compared with the EF group, the NE group can significantly inhibit the amount of albumin leakage in the urine. From this result, it was confirmed that components other than the components of earthworms (Eisenia vulgaris) have the effect of suppressing the leakage of albumin in the urine.

Figures 32 to 35 are related to the measurement results of liver lipids, Figure 32 is the measurement results of liver weight, Figure 33 is the measurement results of liver total lipids, Figure 34 is the measurement results of liver triglycerides, Figure 35 is the total liver The result of the weight measurement of cholesterol.

In the measurement of liver weight, there are significant differences between C group and EF group, and NE group (LSD test, p<0.05), (there is no significant difference between EF group and NE group). In the measurement of total liver lipid, C group, NE group and EF group are significant differences (LSD test, p<0.05) (there is no significant difference between C group and NE group).

In the determination of the weight of triacid anhydride oil in the liver, there are significant differences between the C group, NE group and EF group (LSD test, p<0.05) (there is no significant difference between the C group and the NE group), and the total cholesterol weight measurement In, the results of meaningful differences between groups (LSD test, p<0.05).

Specifically, compared to the C group, the NE group shows the total cholesterol as a meaningful low value, and furthermore, the EF group shows the total cholesterol as a meaningful low value compared to the NE group.

From this result, it has been confirmed that the earthworm (Eisenia vulgaris) ingredient has the effect of suppressing the accumulation of fat in the liver.

Figure 36 is the ECLT measurement result, and Figure 37 is the PAI-1 measurement result.

Same as experiment 1, in this experiment, the ECLT measurement results, group C and There is a significant difference between the EF group (LSD test, p<0.05). Compared with the C group, the EF group shows a low value of ECLT. Moreover, there is no significant difference between the NE group and either the C group or the EF group.

In addition, regarding PAI-1, similar to Experiment 1, there is a significant difference between group C and group EF (LSD test, p<0.01). Compared with group C, group EF shows a lower value of PAI-1. In addition, related The NE group, like the EF group, has a significant difference with the C group (LSD test, p<0.01), and shows a lower PAI-1 value compared to the C group (there is no significant difference between the EF group and the NE group difference).

From the results of the ECLT and PAI-1 measurement, it was confirmed that the earthworm (Eisenia vulgaris) component has the effect of significantly inhibiting the reduction of the fibrinolytic reaction in diabetes, and it was also confirmed that the earthworm (Eisenia vulgaris) component was not included. It has the effect of inhibiting fibrinolysis inhibitor (PAI-1).

Figure 38 shows the proportion of the interstitial matrix in the glomerulus.

Same as experiment 1, there is a significant difference between group C and group EF (LSD test, p<0.05). Compared with group C, group EF shows the area of glomerulus, and the proportion of interstitial stroma in the ring is smaller. That is, it shows that the increase of annulus interstitial matrix is inhibited.

In addition, in this experiment, there was a significant difference between the C group and the NE group (LSD test, p<0.05), and further, there was a significant difference between the NE group and the EF group (LSD test, p<0.05). In other words, the increase in the annulus interstitial matrix in the EF group was suppressed the most, followed by the suppression of the increase in the annulus interstitial matrix in the NE group. From this result, we know that the kidney tissue structure In terms of change inhibition, earthworm components make a great contribution.

From the results of the above test 1 and test 2, it was confirmed that the present composition has the effect of suppressing diabetic complications (improving lipid metabolism and suppressing the progression of diabetic nephropathy).

In addition, regarding the effect of improving liver lipid metabolism, the effect of enhancing fibrinolytic reaction, and the effect of inhibiting changes in renal tissue structure, it was confirmed that the earthworm (Eisenia vulgaris) component in this composition has a great effect, and the effect of improving renal function, The ingredients (notoginseng, fermented ants, fermented Ginkgo biloba extract, fermentation broth) other than the components of earthworms (Eisenia vulgaris) have a great effect.

In summary, this composition is effective in inhibiting fatty liver caused by type 2 diabetes by using the effects of earthworms (earthworms), panax notoginseng, fermented ants, fermented ginkgo biloba extract, and fermentation broth. Kidney hypertrophy is an epoch-making composition that has never been seen before.

In addition to the above-mentioned effects, the composition can exert anti-inflammatory and analgesic effects by the action of fermented ants, and can also be expected to alleviate various diseases caused by diabetes.

In addition, the present invention is not limited to this embodiment, and the specific configuration of each constituent element can be appropriately designed.

Claims (5)

A composition for inhibiting fatty liver caused by diabetes, which is characterized by containing fermented earthworms as an effective ingredient by fermenting the fermentation broth obtained by inoculating yeast in vegetable and fruit extracts. A composition for inhibiting fatty liver caused by diabetes, which is characterized by containing fermented earthworms fermented by the fermentation broth obtained by inoculating yeast in vegetable and fruit extracts as an effective ingredient, and has the ability to inhibit the neutral fat of the liver. The effect of accumulation. The composition for inhibiting fatty liver caused by diabetes according to claim 1 or 2, wherein the aforementioned fermented earthworm is Eisenia vulgaris. A composition for inhibiting kidney hypertrophy caused by diabetes, which is characterized by containing fermented earthworms fermented by the fermentation broth obtained after inoculation of yeast in vegetable and fruit extracts as an effective ingredient, and has a function of enhancing the fibrinolytic reaction effect. The composition for inhibiting kidney hypertrophy caused by diabetes according to claim 4, wherein the aforementioned fermented earthworm is Eisenia vulgaris.

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