KR101168595B1 - A composition comprising an extract of Portulaca oleracea for preventing or treating diabetes, diabetic complications - Google Patents

Abstract

The present invention relates to a composition for the prevention and treatment of diabetes mellitus and diabetic complications containing the extract as an active ingredient. More specifically, in the present invention, the blood glucose lowering effect, the pancreas insulin secretion promotion, vascular inflammation inhibitory effect, vascular endothelial cell function improvement, diabetic kidney function improvement effect, atherosclerotic lesion inhibition effect, etc. The composition containing the extract of Machi Prefecture may be usefully used as a pharmaceutical or health functional food for the prevention and treatment of diabetes and diabetic complications.

March extract, diabetes mellitus, diabetic complications, hypoglycemia, atherosclerosis

Description

A composition comprising an extract of Portulaca oleracea for preventing or treating diabetes, diabetic complications}

The present invention relates to a composition and a nutraceutical composition for the prevention and treatment of diabetes mellitus and diabetic complications disease containing the extract as an active ingredient.

Purslane, a year old with purslane, is commonly found on rural roadsides. Stems and leaves are boiled and eaten as herbs, and since the shape of the leaves is like horse teeth, it is said to be confectionery in Chinese medicine. This large one is not used as a medicine. The small one is tied to the east and dried in the sun for 2-3 days. In Dongbogam, Machi prefecture is cold and tasteless and has no poison to soothe various boils and swellings, faeces, treat boils and injuries caused by iron injuries, and quench thirst. It is said to kill various bugs. It is also used for various diseases such as detoxification, paralysis, diuretics, deadly poisoning, dysentery, swelling, and uterine contractions. The purslane is called sage, and it is said that it is made into powder in Cheongmyeon and Baekye and then eaten in water. Purslane has been mixed with spices and used as a medicinal herb. In the past, folk remedies of ancestors were used as antidote such as detoxification and poisoning. Also, in the Libyan peninsula, antiseptic, anti-scurvy, antispasmodic, diuretic, insect repellent, skin It was also used as a sedative. In addition, studies on muscle relaxation activity and anticancer effects have been reported.

Diabetes mellitus is a representative chronic adult disease. In recent years, the economic development in Korea has led to a rapid increase in the diabetic population due to overeating, lack of exercise, increased stress as well as genetic factors. It is known to suffer from it, and the growth rate of diabetic patients in the future will be further accelerated if proper precautions are not taken, and it is feared that the diabetes crisis will come and become one of the most serious social problems with the aging society.

Diabetes mellitus can be caused by genetic, environmental, and metabolic factors, resulting in decreased insulin secretion in the pancreatic β-cells or hyperglycemia due to insulin resistance in peripheral tissues. Because of the high likelihood of complications such as arteriosclerosis in the blood vessels, the primary goal of diabetes treatment is to lower high blood sugar and maintain normal blood sugar levels. Abnormalities in glucose metabolism will inevitably lead to lipid metabolism. Insulin dysfunction and hyperglycemia lead to abnormal lipid metabolism, which leads to an increase in lipid components in the blood, resulting in impaired normal function of blood vessels, resulting in atherosclerosis. This will occur. In other words, it is a diabetic complication that causes abnormalities caused by arteriosclerosis with systemic blood vessels, and damages the heart and kidneys, resulting in cardiovascular diseases such as angina and myocardial infarction, kidney disease, peripheral necrosis, cataracts, and diabetic retinopathy. Can be.

Recent large prospective and retrospective studies of diabetes mellitus and autopsy have shown that the main cause of death in diabetic patients is coronary artery disease or cerebrovascular disease caused by atherosclerosis, hyperglycemia, hyperinsulinemia, dyslipidemia, hypertension. Because smoking is the main risk factor, the management of risk factors including strict blood sugar control is essential for the prevention and treatment of atherosclerosis.

      Atherosclerosis is an inflammatory lesion that progresses slowly in the arteries and is the main cause of cardiovascular disease. As it is westernized in Korea, diseases such as diabetes, hypertension, and hyperlipidemia are increasing, which leads to an increase in mortality due to cardiovascular disease. Therefore, research on the pathogenesis and treatment of atherosclerosis is very important. Atherosclerosis begins with low-density lipoprotein cholesterol penetrating into vascular endothelial cells and oxidizing, resulting in extravasation of white blood cells or intravascular inflammation, such as the production of cytokines, chemokines, and metalloproteinases. Inflammation is involved at all stages, from the formation of foam cells, to the proliferation and migration of vascular smooth muscle cells, to the formation of plaques and to rupture, but no detailed mechanism of this is known. . Nuclear factor κB (NF-κB), an important transcription factor in the inflammatory response, regulates inflammatory and immune responses, is involved in cell proliferation and apoptosis, and regulates over 160 genes, as well as numerous factors. NF-κB activity is induced by these. In addition, genes regulated by NF-κB as well as factors inducing NF-κB activity are directly or indirectly related to atherosclerosis, and research on this has been actively conducted recently.

 Diabetes is a disease that is difficult to cure but can be controlled. The treatment of diabetes is mainly aimed at preventing the development and progression of diabetic complications caused by poor blood sugar control. Especially, the treatment of type 2 diabetes is first administered by diet or exercise therapy, Oral hypoglycemic drugs are being used. Compounds such as sulfone urea and biguanide are mainly used as hypoglycemic agents as oral hypoglycemic agents, but cause problems such as weight gain, loss of appetite, skin rash, nausea, vomiting, and diarrhea.

Therefore, research and development of alternative medicines using natural products that are safer and effective in preventing and improving diabetes are urgently needed.

Accordingly, the present inventors have made intensive studies on drugs effective in treating diabetes and diabetic complications without side effects, and as a result, the hypoglycemic effect of non-toxic and edible extracts of Machi, the pancreatic insulin secretion, anti-inflammatory action, vascular endothelial cell function The present invention was completed by confirming the effects of improvement, renal function improving effect, and inhibiting atherosclerotic lesion progression.

Therefore, it is an object of the present invention to provide a composition for the prevention and treatment of diabetes and diabetic complication diseases and health functional food composition containing the extract as an active ingredient.

The present invention provides a composition for the prevention and treatment of diabetes mellitus and diabetic complications containing the extract as an active ingredient.

The extract may be prepared by a method of extracting herbal medicine known to those skilled in the art, but is not limited thereto.

The gusset extract,

(a) grinding the dried gusset and extracting with a solvent;

(b) filtering and centrifuging the residue of the extract; And

(c) filtering the supernatant centrifuged; It can be obtained including. In addition, the concentration of the filtrate in the above step may be obtained by further comprising the step of freeze drying.

The gusset extract is more specifically,

In the first step of grinding the dried gusset string, it is immersed for 10 hours to 50 hours, preferably 20 hours to 30 hours in a volume of a third distillation shoe of about 10 to 20 times, preferably 10 times the weight of the dried sample. A second step of hot water extraction by distilling the distilled water containing machi string at 50 ° C. to 150 ° C. for 1 hour to 2 hours; A third step of filtering the residue of the second step by using a gauze and centrifuging at 1000 rpm to 4500 rpm, preferably 2000 rpm to 4000 rpm at 4 ° C. using a centrifuge; A fourth step of taking the separated supernatant by centrifugation and filtering using Whatman No. 3 filter paper; After concentrating this using a vacuum concentrator, it is possible to obtain the Portugese extract of the present invention through a manufacturing method comprising a fifth step of completely freezing at -20 ℃ to freeze-dried in a vacuum over about 2 to 3 days.

The extract of Machi Prefecture of the present invention is preferably extracted with 1 to 30% of the raw material.

According to the experimental example of the present invention, the blood pressure and blood glucose lowering effect, the plasma creatinine concentration and the albumin concentration, the plasma biochemical level improvement effect by the administration of the extract of Macchi in the diabetic animal model db / db mouse, Increasing HDL-cholesterol levels and reducing triglycerides, LDL-cholesterol levels, improving blood lipid abnormalities, vascular function through vasorelaxant effects, and ICAM-1, VCAM-1, E-selectin The inhibitory effect of vascular inflammation was confirmed by a decrease in the expression of the adhesion factor and collagen degrading enzyme MMP-2 and a decrease in the expression of the vasoconstrictor ET-1.

In addition, according to the experimental example of the present invention, diabetes mellitus by reducing the production of the final glycosylated product, decreased expression of transforming growth factor and inhibition of NF-κB p65 activity in the db / db mice of the diabetic animal model according to the administration of the extract Nephrotic inhibitory effect was confirmed.

In addition, according to the experimental example of the present invention, it was confirmed that the effect of increasing insulin secretion according to the administration of the extract of Portugese in the db / db mice of the diabetic animal model.

In addition, according to the experimental example of the present invention, in mice lacking ApoE, which is a typical biomodel that causes atherosclerosis, hepatoprotective effect by reducing the activity of GOT and GPT in plasma according to the administration of extract of Machi, the total in plasma The effect of reducing cholesterol, triglyceride, and LDL-cholesterol levels was found to improve blood lipid abnormalities. In addition, the blood pressure and blood glucose lowering effect, the vascular relaxation effect on acetylcholine and sodium nitropruside, and VCAM-1, a vascular inflammatory factor, were observed in ApoE deficient mice. By reducing the expression of ET-1, the effect of inhibiting vascular inflammation could be confirmed. In addition, the effect of inhibiting vascular atherosclerotic lesions by decreasing expression of MOMA-2, a monosite / macrophage marker, and attenuating lipid staining was observed in ApoE deficient mice.

'Diabetic complication disease' used in the present invention is coronary artery disease such as atherosclerosis, ischemic myocardial infarction, angina pectoris, myocardial infarction, diabetic nephropathy, hypertension, stroke, hyperlipidemia, anemia, migraine, arteriosclerosis, arrhythmia, stroke, hemangioma , Hemangiofibroma or angioplasty and the like.

The present invention also provides a composition for the prevention and treatment of diabetes mellitus and diabetic complications formulated in pharmaceutical unit dosage form by adding a pharmaceutically acceptable carrier, excipient or diluent to the extract.

The pharmaceutical dosage forms of the extracts of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

The composition comprising the extract of Machi, according to the present invention, is formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, oral dosage forms of aerosols, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Can be used.

Carriers, excipients and diluents that may be included in the compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, sucrose ( It is prepared by mixing sucrose or lactose and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate and the like can be used.

As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the active ingredients of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art.

However, for the desired effect, the active ingredient of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg per day. The administration may be carried out once a day or divided into several times. The dosage does not limit the scope of the invention in any aspect.

The active ingredient of the present invention can be administered to mammals such as rats, mice, livestock, humans by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural, or intracerebroventricular injection.

The present invention provides a dietary supplement for the prevention and improvement of diabetes mellitus and diabetic complications disease as an active ingredient extract.

The health functional food of the present invention is a tablet or capsule administered to the human body for the purpose of preventing a specific human disease, alleviating its symptoms, or treating a specific disease, as prescribed by the law (health functional food law). , Pills, liquids, and the like.

The present invention provides a dietary supplement containing Machihyun extract and food acceptable food additives having an effect of preventing and improving diabetes, diabetic complication disease and atherosclerosis.

As defined herein, "food acceptable food additives" are ingredients that are added in trace amounts to foods already known in the art, such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural Flavoring agents such as flavoring agents, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonated drinks It is characterized in that it comprises a carbonation agent and the like used in.

The dietary supplement of the present invention includes forms such as health drinks, tablets, capsules, pills, liquids, and the like.

Examples of the food to which the active ingredient exhibiting prevention and improvement of diabetes and diabetic complication diseases of the present invention can be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods.

It may also be added to food or beverages for the therapeutic and prophylactic purposes pursued by the present invention.

At this time, the amount of the active ingredient in the food or beverage is generally added to the dietary supplement composition of the present invention to 0.01 to 50 weight (%), preferably 0.01 to 15 weight (%) of the total food weight, The beverage composition may be added at a ratio of 0.02 to 5 g, preferably 0.3 to 1 g based on 100 ml.

The health beverage composition of the present invention has no particular limitation except for containing the active ingredient as an essential ingredient in the ratio indicated, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, like ordinary drinks. . Examples of the above-mentioned natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose, for example polysaccharides such as maltose and sucrose, and conventional sugars such as dextrin and cyclodextrin. And sugar alcohols such as xylitol, sorbitol, and erythritol.

As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition, the dietary supplement or composition thereof of the present invention may contain a flesh for preparing natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

The composition containing the extract of Machi Prefecture as an active ingredient has an effect of lowering blood sugar, promoting insulin secretion of the pancreas, inhibiting vascular inflammation, improving vascular endothelial function, improving diabetic kidney function, inhibiting atherosclerotic lesions, and the like. As there is no problem such as toxicity and side effects in the human body, it can be usefully used for the prevention and treatment of diabetes and diabetic complications.

Hereinafter, the examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples in accordance with the gist of the present invention, those skilled in the art to which the present invention pertains. Will be self-evident.

Example 1 Preparation of Machi Prefecture Extract

  2000 ml of distilled water was added to 200 g of Machi prefecture, which was purchased from Kumho-dang (Jeonbuk Jeonju) and powdered with a crusher, and immersed for 24 hours, followed by hot water extraction at 100 ° C. for 2 hours. After cooling, the residue is filtered using gauze, and then centrifuged twice at 4 ° C. and 3000 rpm for 20 minutes using a centrifuge to remove foreign substances that have settled to the bottom, and the separated supernatant is separated into filter paper (Whatman NO. 3) filtered, and then the filtrate was concentrated using a vacuum concentrator, and then lyophilized using a freeze dryer (EYELA FDU-1100) to give 20 g of water extract, the extract was stored at -20 ℃, It was dissolved in purified water and used in the following experimental example.

Reference Example 1 Preparation of a Diabetic Animal Model

  Six-week-old C57BL / 6J-db / db male mice and C57BL / 6J (wild type) male mice, type 2 diabetic animal models, were acclimated for two weeks after purchase from a central laboratory animal. Each group was assigned to have similar mean blood glucose, and the separation of the groups was normal (wild type mouse + water), diabetic control (db / db mouse + water), positive control (db / db mouse + Rosiglitazone) and It was divided into experimental groups (db / db mouse + machi Prefecture water extract administration). All groups underwent a standard AIN-76 diet for 10 weeks, and the diet and drinking water were freely ingested. The temperature and humidity of the nursery were maintained at 20 ~ 25 ℃ and 50 ~ 60%, respectively. The drug was administered to the experimental group at a concentration of 300 mg / kg / day, and the positive control group, Rosiglitazone (Rosiglitazone), which is used as an antidiabetic drug as a PPAR-γ antagonist, was administered at a dose of 10 mg / kg / day. All groups were orally administered for 10 weeks.

Reference Example 2 Preparation of Atherosclerosis Animal Model

The purpose of this study was to evaluate the efficacy of gut extract in inhibiting the development of atherosclerosis in apolipoprotein E knockout mice knocked out of apolipoprotein E, a typical biomodel that causes atherosclerosis. Each group of 6-week-old C57BL / 6J-derived ApoE-/-male mice and C57BL / 6J (wild type) male mice were acclimated for 2 weeks after purchase from central experimental animals. Separation of the group was normal group (wild type mouse + water administration), control group (ApoE-/-mouse + water administration), positive control group (ApoE-/-mouse + captopril administration) and experimental group (ApoE-/-mouse + machihyeonju) Extract administration). The diet and drinking water were freely consumed in all groups, and the temperature and humidity of the feeding room were maintained at 20 to 25 ° C and 50 to 60%, respectively. The drug was administered to the experimental group at 300 mg / kg / day, and the positive control group at 15 mg / kg / for captopril, an ACE inhibitor involved in preventing the development of atherosclerosis. Each group was orally administered for 11 weeks at the dose of day.

<Experiment 1> Confirmation of blood pressure and blood glucose lowering effect according to the administration of Machi extract in db / db mice of type 2 diabetes model

The present inventors conducted the following experiment to determine how the administration of Portulaca oleracea extract on blood pressure and blood glucose in type 2 diabetes model db / db mice.

Figure 1 shows the experimental design to determine the anti-diabetic effect of the extract of Portugese in db / db mice type 2 diabetes model. Each group was treated with rosiglitazone as a positive control in the wild type (C57BL / 6J), db / db control, db / db mice, and in db / db mice. The experiment was carried out by dividing into three groups administered Machi Prefecture extract. Machi water extract (300mg / kg / day) was administered daily for a total of 10 weeks, and sacrifice was sacrificed the day before the autopsy.

The present inventors measured systolic blood pressure every two weeks during the 10-week experimental period to determine the effect of the extract of Machi on the systolic blood pressure of db / db mice. Blood pressure was measured by the tail-cuff method using a blood pressure monitor for animals (Muromachi, Japan). The blood pressure was measured at a constant time (morning) at two-week intervals, and then the white paper was fixed in an animal calibration frame, stabilized at room temperature for 10 minutes, and the average value was calculated by repeating five or more times. Blood glucose was fasted from 6 hours before measurement, and blood was collected from the caudal vein of the mouse and measured using a Onetouch Ultra Blood Glucose Meter and Test Strip. Glucose measurements were measured under the same conditions at regular time intervals over two weeks during the 10-week experimental period.

As shown in Figure 2, the db / db control group showed a significant increase in blood glucose compared to the normal group (wild type), the rosiglitazone administration group showed a significant blood glucose reduction effect compared to the db / db control group. Blood glucose levels were significantly reduced in the group treated with the Machi extract compared to the db / db control. As a result, it was confirmed that the blood glucose lowering effect by the administration of Machi Prefecture extract, and also significant blood pressure lowering effect compared to the control group.

< Experimental Example 2> Confirmation of the improvement of plasma biochemical levels and the improvement of blood lipid abnormalities in the db / db mice of type 2 diabetes model

The present inventors examined the effects of plasma extracts on plasma biochemical changes and blood lipid concentrations in db / db mice, a type 2 diabetes model.

Serum changes due to hyperglycemia showed no significant difference in blood urea nitrogen and total protein levels compared to the control group. The creatinine concentration in plasma was significantly increased in the db / db control group compared to the normal group, and significantly decreased in the rosiglitazone-administered group, and also significantly decreased in the group treated with the present water extract. Plasma albumin concentration was significantly increased in the group treated with Portulaca oleracea compared to db / db control (see FIG. 3).

In addition, as a result of observing changes in blood cholesterol level due to hyperglycemia, HDL-cholesterol levels were significantly decreased in the db / db control group compared to the normal group. Significant increase was observed compared to the db / db control group.

Triglyceride (TG) and LDL-cholesterol levels were significantly increased in the db / db control group compared to the normal group. (See Figure 4).

Experimental Example 3 Confirmation of Vascular Function Improvement and Diabetic Vascular Inflammation Effect in the db / db Mouse of Type 2 Diabetic Model

In order to investigate the improvement of vascular function for the administration of the extract of Machi, the thoracic aorta was extracted and the degree of vascular relaxation was checked. Aceticoline was treated by concentration to analyze the vasorelaxant effect of the gusset extract treated group, and showed significant vasorelaxant effect compared to db / db control group. As a result, it was confirmed that the gut extract administration group improved vascular function depending on vascular endothelial cells. In addition, as a result of treating SNPs acting directly on smooth muscle without concentration through vascular endothelium, it was confirmed that the vascular relaxation effect was significantly higher than that of the db / db control group in the Machi extract extract group, and the vascular relaxation effect was also significant in the Rosiglitazone-administered group. It could be confirmed (see FIG. 5).

In recent years, the expression of cell adhesion molecules has been reported to be the beginning of the detection of vascular complications, and have a great influence on the development of atherosclerosis. In order to analyze the inhibitory effect of vascular inflammation on the extract of Machi, the chest aorta was extracted, and the protein was extracted and western blotting was performed. Adhesion factors such as ICAM-1, VCAM-1, and E-selectin and the expression of MMP-2, a collagen degrading enzyme, were significantly increased in the aorta of the diabetic control group. Decreased. As a result, the gut extract was shown to be effective in inhibiting vascular inflammation by significantly reducing ICAM-1, VCAM-1, and E-selectin expressed in diabetic blood vessels (see FIG. 6).

Immunohistochemical staining was performed on the thoracic aorta of mice to analyze the expression of endothelin-1 (ET-1), a vasoconstrictor. In the normal group, the expression of ET-1 was hardly observed, but it was observed that the expression of ET-1 was increased in the vascular endothelial region of the db / db control group. Reference).

Experimental Example 4 Diabetic Nephropathy Inhibition Effect According to the Administration of Macchi Extract in db / db Mice

The production of advanced glycation end products (AGEs) known to be involved in the development or progression of diabetic nephropathy was analyzed by immunohistochemical staining in the kidney cortex. The decrease was increased, and significantly decreased in the Machi group (see FIG. 8).

Final glycosylation products overexpress various types of cytokines such as transforming growth factor (TGF-β1), which causes tissue fibrosis, causing complications. Expression of TGF-β1 in the kidney cortex by immunohistochemical staining As a result, the expression of TGF-β1 was significantly increased in the diabetic control group compared to the normal group. As a result, it was shown that the extract of Machi can reduce the expression of TGF-β1 in the glomeruli of the diabetic mouse kidney cortex, which may help to suppress diabetic nephropathy (see FIG. 9).

Recent papers have shown that inflammatory responses play an important role in the pathogenesis of diabetic nephropathy. In the present invention, the expression of ICAM-1 involved in the inflammatory response in the kidney tissue was measured by immunohistochemical staining. As a result, the diabetic control group showed a marked increase in expression compared to the normal group. Showed (see FIG. 10).

Western blot and EMSA were used to measure the activity of inflammatory mediator NF-κB p65 in the renal tissues. (See Figure 11).

Experimental Example 5 Confirmation of Increasing Insulin Secretion Effect According to the Administration of Macchi Extract in db / db Mice, a Type 2 Diabetes Model

Immunofluorescence staining was performed on the pancreas frozen sections of mice to analyze the expression of insulin in the pancreas islet, a tissue that secretes insulin in the pancreas. In the normal group, the expression of insulin in the pancreatic islets was increased, and in the db / db control group, the expression of insulin was significantly decreased due to the necrosis of β-cell. The rosiglitazone-administered group showed a significant expression increase compared to the db / db control group, and the gusset extract-treated group showed a significant expression increase. As a result, it was confirmed that the gut water extract improves the insulin secretion ability of the pancreas (see FIG. 12).

Experimental Example 6 Confirmation of Plasma Biochemical Level and Plasma Lipid Abnormalities in ApoE-/-Mouse

In 1992, apoE-deficient mice were made at the University of North Carolina in the United States using gene knockout technology, which not only produces fat streak, but also has an overall similarity in human blood vessels. Fibrous caps also form, such lesions include the base of the aorta, the small part of the thoracic aorta, the carotid artery, the intercostal artery, the mesenteric artery, the renal artery, the branch of the iliac artery, and the proximal coronary, carotid, femoral, It has been reported to form in the subclavian artery and the clavicle artery. In addition, since the progression of lesions is accelerated as the high cholesterol diet is performed, the mice are used as genetic animal models that can cause atherosclerosis and arteriosclerosis.

In apolipoprotein E knockout mouse, a typical biomodel inducing atherosclerosis, the present inventors examined the effects of gut extract on plasma biochemical changes and blood lipid concentrations as follows. The experiment was conducted.

 The inventors of the present invention have examined the inhibitory effect of atherosclerosis of water extracts of Machise in ApoE-/-mice.

 Six-week-old C57BL / 6J-derived ApoE-/-male mice and C57BL / 6J (wild type) male mice were acclimated for two weeks, and each group was assigned to have the same average weight. Mice + water administration), control group (ApoE-/-mouse + water administration), positive control (ApoE-/-mouse + captopril administration) and experimental group (ApoE-/-mouse + Machi Prefecture water extract administration). In the experimental group, Machise water extract was administered at a concentration of 300 mg / kg / day, and in the positive control group, a dose of 15 mg / kg / day was captopril, an ACE inhibitor involved in preventing the progression of atherosclerosis. Each group was orally administered for 11 weeks (see FIG. 13).

First of all, the activity of GOT and GPT in plasma was measured to determine the changes in liver function, especially toxic effects of Machi prefecture. It showed an effect. That is, it was confirmed that the extract of Machi has no toxicity and showed a protective effect on liver function (see FIG. 14).

In addition, the plasma levels of total cholesterol, triglycerides and LDL-cholesterol were significantly increased in the control group compared to the normal group, and it was confirmed that significant reduction effect was obtained by the administration of choroid. In other words, it was confirmed that the lipid lipid abnormality improving effect in ApoE-/-mice due to the administration of water extract of Machi (see FIG. 15).

<Experiment 7> Confirmation of blood pressure lowering effect, improvement of vascular function and inhibition of vascular inflammation according to the administration of Machi extract in ApoE-/-mouse

During the 11-week experimental period, the control group showed a significant increase in blood pressure and blood glucose, compared with the normal group, and was significantly decreased by the administration of port gut. Figure 16 shows the blood pressure and blood glucose lowering effect according to the administration of the water extract of the portray in ApoE-/-mice.

In addition, in the thoracic aorta isolated from the control group of 19 weeks old, the vasorelaxant effect on acetylcholine was significantly decreased compared to the normal group, but was significantly improved by administration of gut string. In addition, the vasorelaxant effect on sodium nitroprusside was also significantly improved by the administration of port guts. Figure 17 shows the effect of vascular relaxation according to the administration of water extracts from the ApoE-/-mice.

In addition, Western blots and immunohistochemical staining were used to analyze the expression of vascular inflammation factor VCAM-1 in vascular tissues. . Figure 18 shows the effect of inhibiting VCAM-1 expression by the administration of Portulaca extract in the thoracic aorta of ApoE-/-mice.

In addition, the analysis of ET-1 expression in vascular tissues using immunohistochemical staining showed a significant increase in expression in the control group compared to the normal group, and showed a significant decrease in expression in the administration group (FIG. 19). Reference).

Experimental Example 8 Confirmation of Inhibition of Atherosclerotic Lesions in Vascular Aqueous Extracts Treated with Portage Extracts in ApoE-/-Mice

As a result of observing the expression level of MOMA-2 in the atherosclerotic lesions of blood vessels, the expression of MOMA-2 was significantly increased in the control group and markedly decreased in the administration group (see FIG. 20).

In addition, as a result of observing the expression level of nitrotyrosine, which is a marker of oxidative stress, it was significantly increased in the control group compared to the normal group, and effectively decreased in the administration group as shown in FIG.

In addition, oil red O staining was performed to observe lipids in atherosclerotic lesions of blood vessels. Histologically comparing the atherosclerotic lesions, the incidence of atherosclerotic lesions was very low in the Machi group. As can be seen in FIG. 22, in the thoracic aorta of ApoE − / − mice, lipid precipitation was inhibited by administration of Portage water extract (see FIG. 22).

Figure 1 shows an experimental design for determining the anti-diabetic effect of the extract of Portugese in db / db mice type 2 diabetes model.

Figure 2 shows the blood pressure and blood sugar lowering effect according to the administration of Portulae extract in db / db mice.

Figure 3 shows the effect of improving the plasma biochemical levels in the db / db mice according to the administration of Portuguese extract

Figure 4 shows the effect of improving the lipid lipids in the blood according to the administration of the gut extract in db / db mice.

Figure 5 shows the effect of vascular relaxation using the thoracic aorta in order to find out the improvement of vascular function according to the administration of choroid extract in db / db mice.

FIG. 6 illustrates Western blotting using the thoracic aorta to analyze the inhibitory effect of vascular inflammation upon administration of Portulaca oleracea extract in db / db mice. ICAM-1, VCAM-1, E involved in vascular inflammation Adhesion factors such as -selectin and MMP-2, a collagen degrading enzyme, have been shown to reduce expression.

Figure 7 shows the effect of immunohistochemical staining in the thoracic aorta of db / db mice, showing the effect of inhibiting the expression of Endothelin-1 (ET-1) by administration of Portulaca oleracea extract.

Figure 8 shows the inhibitory effect of the production of advanced glycation end products (AGEs) by administering the immunohistochemical staining method in the kidney cortex of db / db mice.

Figure 9 shows the effect of inhibiting the expression of transforming growth factor (TGF) -β1 by administration of the extract of the gut by performing immunohistochemical staining in glomeruli in the kidney cortex of db / db mice.

Figure 10 shows the effect of inhibiting the expression of ICAM-1 by the administration of the extract of Portugese measured by immunohistochemical staining in the kidney tissue of db / db mice.

Figure 11 shows the effect of inhibiting the NF-κB p65 activity by the administration of the extract of the chorus, using Western blot and electrophoretic mobility shift assay (EMSA) in the kidney tissue of db / db mice.

Figure 12 shows the effect of increasing the secretion of insulin by the administration of cholangi extract in pancreas islet, which is a tissue that secretes insulin in the pancreas, by performing immunofluorescence staining on frozen pancreatic sections of db / db mice.

Figure 13 shows the experimental design for determining the atherosclerosis inhibitory effect of Portugese extract in ApoE-/-mice.

Figure 14 shows the protective effect of liver function through the decrease in the activity of GOT, GPT in plasma according to the administration of gut extract in ApoE-/-mice.

Figure 15 shows the effect of improving the lipid lipids according to the administration of the gut extract in ApoE-/-mice.

Figure 16 shows the blood pressure and blood glucose lowering effect according to the administration of the Portugese extract in ApoE-/-mice.

Figure 17 shows the effect of vascular relaxation according to the administration of the gut extract in ApoE-/-mice.

Figure 18 shows the effect of inhibiting the expression of VCAM-1 by the administration of the chord extract in the thoracic aorta of ApoE-/-mice using Western blot and immunohistochemical staining.

19 shows the effect of inhibiting the expression of ET-1 by the administration of the extract of Machi in the thoracic aorta of ApoE − / − mice using immunohistochemical staining.

20 shows the effect of inhibiting the expression of MOMA-2 by the administration of chordal extract in the thoracic aorta of ApoE − / − mice using immunohistochemical staining.

FIG. 21 shows the effect of nitrotyrosine expression on the thoracic aorta of ApoE − / − mice by immunohistochemical staining.

Figure 22 was carried out Oil Red O staining to observe the lipids in atherosclerotic lesions of blood vessels in ApoE-/-mice, showing the effect of inhibiting lipid precipitation by the administration of Portulaca extract.

Claims (10)

Atherosclerosis preventive and therapeutic pharmaceutical composition containing water extract as an active ingredient. The water extract of claim 1, wherein (a) grinding the dried gusset string and extracting with water; (b) filtering and centrifuging the debris of the water extract of Portage of step (a); And (c) filtering the supernatant centrifuged in step (b); It is obtained by, Atherosclerosis prevention and treatment pharmaceutical composition. delete delete The composition of claim 1, wherein the composition is formulated in a pharmaceutical unit dosage form with the addition of a pharmaceutically acceptable carrier, excipient or diluent. The method of claim 5, wherein the pharmaceutical unit dosage form is selected from the group consisting of powders, granules, tablets, capsules, suspensions, emulsions, syrups, oral formulations of aerosols, external preparations, suppositories and sterile injectable solutions. Composition. 7. A composition according to claim 5 or 6, wherein said pharmaceutical unit dosage form is formulated to be able to administer portage water extract in an amount of 0.0001 to 100 mg / kg body weight per day. Health functional food for prevention and improvement of atherosclerosis, which contains water extract of Machi as an active ingredient. The health functional food according to claim 8, wherein the form of the health functional food is selected from the group consisting of tablets, capsules, pills, and liquids. 10. The health functional food according to claim 8 or 9, wherein the health functional food comprises 0.001 to 50% of water extract of Machi based on the total weight of the raw material.

Images