KR100525816B1 - An antidiabetic composition comprising O. japonicus polysaccharide extract - Google Patents

Abstract

The present invention relates to an antidiabetic composition comprising an extract of polysaccharides, and the blood of experimental animals bred as described above after breeding by administering an experimental diet comprising the extract to a diabetic-induced experimental animal after obtaining the capsules. It is very useful in the food and pharmaceutical industry because it can provide an antidiabetic composition containing the extract of polysaccharides by measuring the lipid content of blood sugar and serum by extracting.

Description

Antidiabetic composition comprising O. japonicus polysaccharide extract

The present invention relates to an antidiabetic composition comprising a polysaccharide extract of Warsong, and more particularly, to an antidiabetic composition containing a polysaccharide extract extracted from Korean Wasong as an active ingredient.

Wasong is a perennial herbaceous plant of Crassulaceae, also known as Orostachys japonicus A. Berger, also known as Amsong, Oksong, and Leafy Herb. It is used for fever, swelling, hemostasis, and diarrhea in Korea. (金 在 佶, 1984, 原色 天然 藥物 大事 典 (上 卷), 南山 堂, Seoul, p.447) 1990, Oriental Medicine and Health, 1, 26].

This plant is a fleshy perennial herb that grows by the side of a rock, but it is characterized by flowering and fruiting. The rosettes are rosette-like, and the ends are hardened and become like thorns. The leaves are in the main stem, there are no foliar leaves, and the rosettes coming out in the summer are not stiffened and just pointed. have. Flowers bloom in September, white, total inflorescence is 6-15cm long, without flowers, close to the flowers, lanceolate, and pointed. 5 sepals are lanceolate, green, 5 petals, lanceolate acute, 6mm long. There are 10 stamens, longer than petals, 5 ovaries, and pots are red, but gradually become black [Lee Chang-bok, 1979, 大 韓 植物 圖鑑, 鄕 文 社 Seoul, p.402]. Wasong is harvested from summer to autumn, and dried root-extracted vinegar is medicinal [金 在 佶, 1984, 原色 天然 藥物 大事 典 (上 卷), Namsan Mountain, Seoul, p.447]. .

If you look through the literature through the literature, it is said that Dangboncho is used for dryness and pain in the mouth. In the herbal herb, it is extracted with water in colonic hemorrhage and applied to all wounds. It is said to have a converging effect. Herbal rehabilitation treats white poison and kills the species, and classifies herbicides to treat hemorrhoidal hemorrhage and to wash the herb in full amount. It is said to have an effect on 과 and [[上海 科學 技術 出版社 小學 館 編;中藥 大事 典, 小學 館, 1985, p.526].

Soluble components reported so far are triterpenoids such as priedelin, epi-friedlanol, glutinone, and glutinol. And sterol-based materials such as β-sitosterol and campesterol, fatty acid esters and flavonoids such as kaempferol and quercetin. aromatic acids such as (flavonoid), 4-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid, and gallic acid. Park, JG, Park, JC, Hur, JM, Park, SJ, Choi, DR, Shin, DY, Park. K. Y., Cho, H. W. and Kim, M. S., 2000, Phenolic compounds from Orostachys japonicus having anti-HIV-1 protease activity, Nat. Prod. Sci. 6 (3), p. 117 to 121; Park, H. J., S. C., Lee, M. S. and Young, H. S., 1994, Triterpene and steroids from Orostachys japonicus, Kor. J. Pharmacog. 25 (1), p. 20-24; Park, H. J., Young, H. S., Kim, J. O., Rhee, S. H. and Choi, J. S., 1991a, A study on the chemical constituents of Orostachys japonicus A. Berger, Kor. J. Pharmacog. 22 (2), p. 78-84; Park, H. J., Young, H. S., Park, K. Y., Rhee, S. H., Chung, H. Y. and Choi, J. S., 1991b, Flavonoids from the whole plants of Orostachys japonicus, Arch. Pharm. Res. 14 (2), p. 167-171]. In addition, seopheptulosan, isopropylidene sedoheptulose, and flavonoids are known to be used as sugar components in O. fimbriatus Berger from China. Kaempferol-3-glucosyl-7-rhamnoside is known [左 春旭, 英 英, 姜岩 靑, 蔡玉英, 丁 杏 包, 1988, 瓦松 中, 藥草, , 19, 148;左 春旭, 蔡玉英, 姜岩 靑, 王建國, 1985, 瓦松 有效 成分 的 硏 究, 中草藥, 16, 243]. Some studies on the composition of these vortices have been reported, but no report on their biological activity has been systematically verified in Korea.

Therefore, the present inventors extract the polysaccharide fraction from Korean Wasong, and administer the experimental diet including the polysaccharide extract to a diabetic-induced experimental animal, and then collect the blood of the experimental animal bred as described above to obtain blood glucose and lipid in serum. An object of the present invention is to provide an antidiabetic composition containing a polysaccharide extract isolated from vortex by measuring the content.

Accordingly, an object of the present invention is to extract the polysaccharide fraction from the Korean Waksong, and to raise the blood glucose and the blood of the experimental animals bred as described above after breeding by administering the experimental diet comprising the polysaccharide extract to the diabetes-induced experimental animals By measuring the lipid content in the serum to provide an anti-diabetic composition containing the polysaccharide extract.

The object of the present invention is to obtain a Wosong polysaccharide extract, and then to breed the experimental diet containing the polysaccharide extract to the diabetic-induced experimental animals, and to collect blood of the experimental animals bred as described above, the blood glucose and serum lipid content It was found that the Warson polysaccharide extract was antidiabetic and completed the present invention.

Hereinafter, the configuration of the present invention.

The present invention comprises the steps of extracting the polysaccharide fraction from the vortex; Preliminary breeding of the experimental animals to induce diabetes, and administering the experimental diets including the Warson polysaccharide extract and the control diet not containing the diabetic-induced experimental animals; Comprising the blood of the experimental animal of the above step and measuring the lipid content in blood sugar and serum.

In order to achieve the above object, the present invention concentrates the hot-water extract of Wahsong, and adds ethanol to the concentrate, and is left at 4 ° C. for 24 hours, followed by centrifugation at 5,000 rpm for 20 minutes to produce a Warsong polysaccharide extract having an anti-diabetic effect. Provide a way to separate.

Whippong used in the present invention is used to finely crush dry flowers or stems.

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Hereinafter, the present invention will be described in more detail with reference to the following Examples and Experimental Examples. However, the following Examples and Experimental Examples are only for illustrating the present invention, the scope of the present invention is not limited to these.

Example 1 Polysaccharide Fraction Extraction

The dried flowers and stems of Orostachys japonicus A. Berger were finely pulverized, and 100 g of the hot water extract was obtained by high pressure (DW 2L, 3 hours, 100 ° C). The extract was filtered with filter paper (whatman NO.1) and concentrated under reduced pressure with a rotavapor (BCHI R-205) to 1/10 the volume. Ethanol was added to the concentrated extract so that the final concentration of ethanol was 85%, left at 4 ° C. for 24 hours, followed by centrifugation (5,000 rpm, rotor 9, 20 min, 18 ° C.) to obtain a supernatant . 12% of japonicus polysaccharide extract (OJP1) was obtained.

Experimental Example 1 Breeding Experiment Animals and Inducing Diabetes

Breeding experimental animals

Seven-week old Sprague-Dawley male rats weighing 200-230 g were purchased and pre- bred with solid feed for a week. After the acclimatization period, diabetes was induced and divided into a normal control group (Normal-Cont), a diabetic experimental group (diabetic control group (Db-Cont), and Wahson group (Db-OJP1)) in consideration of body weight and blood glucose concentration. During the experimental period, the diet and water of the experimental animals were freely taken (ad libitum). Feed intake and body weight were measured at weekly intervals. The breeding conditions of the experimental animals were controlled at room temperature of 22 ± 2 ℃, humidity of 55 ± 5%, and contrast for 12 hours (day light 06: 00-18: 00).

Experimental diet

For animal experiments, the normal control and diabetic control group used powdered feed and the Wasong group used 1% OJP1.

Diabetes

Diabetes induced streptozotocin, which is known to cause hyperglycemia due to insulin deficiency within 24 hours after administration, destroying by acting only on beta-cells of the pancreas without affecting other organs after one week of preliminary breeding. ) (STZ, Sigma Chemical Co., St. Louise, USA) was used. Dissolved in 0.01M citric acid buffer (pH 4.5) and intraperitoneally injected at a concentration of 55mg / kg, fasted 12 hours after 3 days, and collected from the tail vein to collect blood glucose levels above 200mg / dL. Used.

Experimental Example 2 Biochemical Analysis of Blood of Experimental Animals after Administration of Wasong Extract

Blood collection

Initial blood was collected from the tail vein after fasting the animals 12 hours on the first day of the experiment. After the animals were fasted for 12 hours on the last day of 4 weeks of breeding, 300 mg / mL urethane (Sigma Chemical Co., St. Louise, U.S.A) was injected into the abdominal cavity for anesthesia and sacrificed. Blood was collected from the abdominal vena cava of experimental animals, and centrifuged (6,000 rpm, 4 ° C., 15 min) to separate serum from the supernatant and stored frozen.

Blood sugar measurement

Blood glucose was measured before starting the experimental diet and on the last day of the experiment. Blood glucose was measured by glucose oxidase method (Sigma Chemical Co., St. Louise, USA), and glucose / urea nitrogen (Sigma Chemical Co., St. Louise) as a standard solution , USA). Absorbance was measured using a spectrophotometer with a blank as a control at 505 nm.

Determination of Lipid Content in Serum

The triglyceride content in serum was measured using a kit reagent (Asan Pharm. Co. Korea) for quantification by enzyme method. That is, enzyme reagent [lipoprotein lipase 10800U, glycerol kinase 5.4U, peroxidase 135000U, L-α-glycero phosphooxidase 160U ] Is dissolved in an enzyme reagent solution [containing 0.427 g / LN, N-bis (hydroxyethyl) -2-aminoethane sulfonic acid (containing N, N-bis (hydroxyethyl) -2-aminomethane sulfonic acid)]. After the addition of 3.0 mL of the enzyme solution prepared in serum and standard solution (Asan Pharm. Co.) and incubated at 37 ° C. for 10 minutes, the absorbance was measured at 550 nm using a spectrophotometer with a blank reagent. The triglyceride content was calculated by this.

The total cholesterol content in serum was measured using a kit reagent (Asan Pharm. Co. Korea) for quantification by enzyme method. That is, the enzyme reagent (cholesterol esterase 20.5KU / L, cholesterol oxidase 10.7KU / L, sodium hydroxide 1.81g / L) buffered [basic potassium phosphate ( potassium phosphate monobasic; KH ₂ PO ₄ 13.6g / L, containing 1.88g / L phenol] and 3.0mL of enzyme solution prepared in 20µl of serum and standard solution (Asan Pharm. Co. Korea), respectively. After incubation at 37 ° C. for 5 minutes, the total cholesterol content was calculated by measuring the absorbance at 500 nm using a spectrophotometer as a control for the reagent blank.

The HDL-cholesterol content in the serum was measured using a kit reagent (Asan Pharm. Co. Korea) for quantification by enzyme method. That is, 0.2 mL of serum was added 0.2 mL of the settling reagent (containing 5 g / L of sodium phosphotungstate and 10 g / L of sodium chloride), mixed well, and left at room temperature for 10 minutes, followed by centrifugation. Take 0.1 mL of the supernatant and buffer the enzyme reagent (containing 20.5 KU / L of cholesterol esterase, 10.7 KU / L of cholesterol oxidase, 1.81 g / L of sodium hydroxide). After mixing well with 3.0 mL of enzyme solution prepared by dissolving in 13.6 g / L potassium phosphate and 1.88 g / L phenol), incubating at 37 ° C. for 5 minutes, and using a spectrophotometer as a control The HDL-cholesterol content was calculated by measuring the absorbance at 500 nm.

The antidiabetic effect was confirmed by supplying 1% OJP1 for 4 weeks to SD diabetic rats induced with Streptozotocin. Plasma glucose, serum triglyceride, serum triglyceride, Serum total cholesterol level was significantly reduced than in the diabetic control group (Table 2, Figures 1-4). Serum HDL-cholesterol levels showed an increasing trend compared to the diabetic control group (p = 0.083). Therefore, it was confirmed that OJP1 is effective in lowering blood sugar, lowering triglyceride and lowering cholesterol in diabetic rats.

Weight Changes in Diabetic Rats Who Take OJP1 for 4 Weeks Group (n) Weight (g) Early In a week 2 weeks later 3 weeks later 4 weeks later Normal control 256.3 ± 2.0 295.2 ± 2.8 325.4 ± 4.3 353.1 ± 4.8 373.1 ± 5.3 Diabetic experimental group (diabetic control group) 224.8 ± 2.4 210.7 ± 5.8 209.8 ± 7.1 208.4 ± 7.0 203.4 ± 7.5 Wasong-gun 238.9 ± 4.2 231.3 ± 11.8 224.3 ± 10.7 219.0 ± 11.1 218.1 ± 13.6

[Note] The weight change was -21.4 ± 6.6g in the diabetic group and -20.8 ± 10.2g in the diabetic group.

Blood Glucose, Serum Triglycerides, Serum Total Cholesterol Levels, and Serum HDL-Cholesterol Levels in Diabetic Rats Who Take OJP1 for 4 Weeks Volume (mg / dL) group Normal control Diabetes experimental group Wasong-gun Blood sugar Before the experiment 94.6 ± 5.9 285.2 ± 25.5 375.9 ± 21.1 After the experiment 95.4 ± 2.2 481.1 ± 15.2 ^† 467.7 ± 8.4 ^† Serum triglycerides Before the experiment 29.6 ± 1.3 94.7 ± 13.8 113.6 ± 16.5 After the experiment 51.3 ± 6.6 638.0 ± 94.6 ^† 240.6 ± 22.4 ^† Serum total cholesterol levels Before the experiment 88.1 ± 2.1 68.8 ± 4.2 75.6 ± 4.9 After the experiment 94.2 ± 4.3 136.2 ± 13.6 ^† 103.4 ± 9.6 ^† Serum HDL-Cholesterol Levels Before the experiment 34.8 ± 0.9 35.4 ± 1.7 35.6 ± 1.5 After the experiment 32.7 ± 2.4 23.8 ± 2.3 ^† 32.3 ± 2.4 ^†

As described above, the present invention relates to an anti-diabetic composition comprising a polysaccharide extract of a hawthorn, and extracts an extract containing a polysaccharide fraction from a songsong and then comprises an experimental diet comprising the polysaccharide extract in a diabetic-induced experimental animal. After the administration and breeding, the blood of experimental animals bred as described above is collected and the lipid content of the blood sugar and serum can be measured to provide an anti-diabetic composition containing the polysaccharide extract of the hawthorn, which is very useful for the food and pharmaceutical industries.

Figure 1 is a graph showing the change in blood glucose of diabetic rats ingested Waksong polysaccharide extract OJP1 of the present invention for 4 weeks. Here, † indicates the significant difference between the diabetic experimental group and the Wasson group was P <0.05 when the Student's Tee test was used.

Figure 2 is a graph showing the change in serum triglycerides of diabetic rats ingested Waksong polysaccharide extract OJP1 of the present invention for 4 weeks. Here, † indicates the significant difference between the diabetic experimental group and the Wasson group was P <0.05 when the Student's Tee test was used.

Figure 3 is a graph showing the change in serum total cholesterol of diabetic rats ingested the present Waksong polysaccharide extract OJP1 for 4 weeks. Here, † indicates the significant difference between the diabetic experimental group and the Wasson group was P <0.05 when the Student's Tee test was used.

Figure 4 is a graph showing the change in serum HDL-cholesterol of diabetic rats ingested Waksong polysaccharide extract OJP1 of the present invention for 4 weeks. Here, † indicates the significant difference between the diabetic experimental group and the Wasson group was P <0.05 when the Student's Tee test was used.

Claims (2)

2 liters of distilled water was added to 100 g of vortex, and hot water was extracted at 100 ° C. for 3 hours. The hot water extract obtained above was concentrated under reduced pressure to 1/10 of the volume, Ethanol was added to the concentrate so that the final concentration of ethanol was 85%, and left at 4 ° C. for 24 hours, Centrifugation at 5,000 rpm for 20 minutes to produce a supernatant with anti-diabetic effect, comprising the step of obtaining a supernatant. Antidiabetic composition, characterized in that it contains as an active ingredient the extract of polysaccharides obtained according to the method of claim 1.