BOWEL MOVEMENT IMPROVING COMPOSITION COMPRISING TENEBRIONIDAE LARVA EXTRACT

Abstract

The present invention relates to a composition comprising a Tenebrionidae larva extract for improving a bowel movement function.

Claims

1. A composition comprising a solvent extract of Tenebrionidae larva for improving a bowel movement function.

2. The composition of claim 1, wherein the solvent is water, an organic solvent, or a mixed solvent thereof.

3. The composition of claim 2, wherein the organic solvent is alcohol having 1 to 4 carbon atoms.

4. The composition of claim 1, wherein the improvement of the bowel movement function is an increase in fecal weight, an increase in water content in feces, an increase in gastrointestinal transport capacity, or an increase in length and area of intestinal mucosa.

5. A composition comprising a solvent extract of Tenebrionidae larva for improving constipation.

6. The composition of claim 5, wherein the constipation is transient constipation, atonic constipation, spastic constipation, bowel movement impaired constipation, or organic constipation.

7. A health functional food comprising a solvent extract of Tenebrionidae larva for improving constipation.

8. A method of improving a bowel movement function in a subject in need thereof comprising administering an effective amount of the composition according to claim 1 to the subject.

9. The method of claim 8, wherein the solvent is water, an organic solvent, or a mixed solvent thereof.

10. The method of claim 9, wherein the organic solvent is alcohol having 1 to 4 carbon atoms.

11. The method of claim 8, wherein the improvement of the bowel movement function is an increase in fecal weight, an increase in water content in feces, an increase in gastrointestinal transport capacity, or an increase in length and area of intestinal mucosa.

12. A method of improving constipation in a subject in need thereof comprising administering an effective amount of the composition according to claim 5 to the subject.

13. The method of claim 12, wherein the constipation is transient constipation, atonic constipation, spastic constipation, bowel movement impaired constipation, or organic constipation.

Description

DESCRIPTION OF DRAWINGS

[0098] FIG. 1 is a diagram illustrating results of measuring fecal weight and water content in feces to determine an effect of administration of a Tenebrionidae larva extract on a fecal index. [N (Normal): normal control; C (Control): loperamide alone administered group; PC (Positive control): Psyllium husk 120 mg/kg; M12: loperamide+Tenebrionidae larva enzyme extract 120 mg/kg; M500: loperamide+Tenebrionidae larva enzyme extract 500 mg/kg; M′120: loperamide+Tenebrionidae larva ethanol extract 120 mg/kg, an error bar represents a standard error, and n=5]

[0099] FIG. 2 is a diagram illustrating results of measuring digestive tract mobility to determine an effect of administration of a Tenebrionidae larva extract on a gastrointestinal transport capacity. [N (Normal): normal control; C (Control): loperamide alone administered group; PC (Positive control): Psyllium husk 120 mg/kg; M120: loperamide+Tenebrionidae larva enzyme extract 120 mg/kg; M500: loperamide+Tenebrionidae larva enzyme extract 500 mg/kg; M′120: loperamide+Tenebrionidae larva ethanol extract 120 mg/kg, an error bar represents a standard error, and n=5]

[0100] FIG. 3 is a diagram illustrating results of staining a colon tissue with hematoxylin-eosin to determine an effect of administration of a Tenebrionidae larva extract on intestinal digestive movement.

BEST MODE

[0101] Hereinafter, the present invention will be described in detail by Examples and Experimental Examples However, these Examples and Experimental Examples are only illustrative the present invention, and the scope of the present invention is not limited to these Examples and Experimental Examples.

PREPARATION EXAMPLE 1

[0102] Preparation of Tenebrionidae Larva Enzyme Extract

[0103] (1) Step of Controlling Diet of Tenebrio molitor Larva

[0104] 3-month-old Tenebrio molitor larva dieted on wheat Grains was subjected to a process of excreting an excrement while fasting before use. The Tenebrio molitor larva after 48-hour fasting was washed and frozen-stored.

[0105] (2) Step of Hydrolyzing, Sterilizing, and Wet-Crashing Tenebrio molitor Larva

[0106] 100 parts by weight of the pretreated raw material was added with 1900 parts by weight of water and then sterilized at 100° C. for 30 minutes. Thereafter, the Tenebrio molitor larva was crushed with a homogenizer at 6000 rpm for 10 minutes to obtain a liquid form that was easily treated with enzymes.

[0107] (3) Step of Treating Tenebrio molitor Larva Solution with Protease to Decompose Protein

[0108] The crushed Tenebrio molitor larva solution was added with 0.1% of protease, Protamex, and treated at a temperature of 60° C. for 1 hour and then the enzyme was inactivated at 100° C. for 10 minutes.

[0109] (4) Step of Filtering Tenebrio molitor Larva Extract

[0110] The protease-treated product was filtered under reduced pressure through 1 μm glass fiber filter paper, and then only a filtrate was taken.

[0111] (5) Step of Concentrating Tenebrio molitor Larva Extract

[0112] The filtrate was concentrated with a vacuum concentrator to prepare a concentrate of 10 Brix or more.

[0113] (6) Step of Powdering Tenebrio molitor Larva Extract

[0114] The concentrate was lyophilized and powdered, and specifically, sequentially maintained at −45° C. for 6 hours, at −20° C. for 21 hours, at −15° C. for 11 hours, and at —5° C. for 11 hours and then thawed, and then powdered by removing moisture.

PREPARATION EXAMPLE 2

[0115] Preparation of Tenebrionidae larva ethanol extract

[0116] (1) Step of Controlling Diet of Tenebrio molitor Larva

[0117] 3-month-old Tenebrio molitor larva dieted on wheat grains was subjected to a process of excreting an excrement while fasting before use. The Tenebrio molitor larva after 48-hour fasting was washed and frozen-stored.

[0118] (2) Step of Drying and then Crushing Tenebrio molitor larva

[0119] The frozen-stored Tenebrio molitor larva was washed and then dried at 90° C. for 24 hours. The dried raw material was finely crushed with a crusher for easy extraction.

[0120] (3) Step of Extracting Tenebrio molitor Larva Powder by Adding Ethanol

[0121] 100 parts by weight of the pretreated raw material was added with 900 parts by weight of ethanol and then extracted at 25° C. and 100 rpm for 24 hours.

[0122] (4) Step of Filtering Tenebrio molitor Larva Extract

[0123] The extract was filtered under reduced pressure through 1 μm glass fiber filter paper, and then only a filtrate was taken.

[0124] (5) Step of Concentrating Tenebrio molitor Larva Extract

[0125] The filtered extract was concentrated with a decompression concentrator to prepare a concentrate of 10 brix or more.

[0126] (6) Step of Powdering Tenebrio molitor Larva Extract

[0127] The concentrate was lyophilized and powdered, and specifically, sequentially maintained at −45° C. for 6 hours, at −20° C. for 21 hours, at −15° C. for 11 hours, and at −5° C. for Ii hours and then thawed, and then powdered by removing moisture.

EXPERIMENTAL EXAMPLE

[0128] Confirmation of Bowel Movement Function of Tenebrionidae Larva Extract

Experiment Example 1: Breeding of Experimental Animals and Design of Experiments

[0129] This study was conducted in accordance with the policies and regulations of the Institutional Animal Care and Use committee (SEMI-19-008) of Southeast Medi-Chem Institute. As experimental animals used in the experiment, a 7-week-old ICR mouse model (male) was received from Maria Sic (Seongnam, Gyeonggi-do, Korea) and quarantined, acclimatized and bred in an animal kennel (Animal Facility Registration Certificate: No. 412) and then experimented. During breeding, a lighting time was set to a 12-hour (07:00 to 19:00) cycle, and a diet and water were freely ingested.

[0130] First, 5 ICR mice (7 weeks old, male) were placed in each group, and the experimental groups were classified into a total of 6 groups. The experimental groups were divided into a normal control group (N), a loperamide administered group (C), a Psyllium husk administered group (PC), a loperamide+low concentration enzyme extract group (120 mg/kg) (M120), a loperamide+high concentration enzyme extract group (500 mg/kg) (M500), and a loperamide+low concentration ethanol extract group (120 mg/kg) (M′120).

[0131] The samples were orally administered for a total of 7 days according to the contents in Table 1, and on day 7 of administration, all the groups except for the normal control group were orally administered with 5 mg/kg of loperamide (in saline) to induce constipation. For a feed, solid feed for the experimental animals (Rana Bio, Seongnam, Gyeonggi-do, Korea) was provided, and changes in body weight was measured twice a week.

TABLE-US-00001 TABLE 1 Group Experiment Note Control group 1 (N) Administration of No administration saline of Loperamide Control group 2 (N) Saline Control group 3 (PC) Psyllium husk 120 mg/kg Example 1 (M120) Preparation Example 1 120 mg/kg Example 2 (M500) Preparation Example 1 500 mg/kg Example 3 (M′120) Preparation Example 2 120 mg/kg

[0132] The measured experimental results were represented as means and standard errors, and the significance test of each group was statistically processed using an anova t-test in the Statview program.

Experiment Example 2: Changes in Fecal Index by Tenebrionidae Larva Extract

[0133] The feces of the experimental animals were collected after administration of loperamide (5 mg/kg, body weight) on day 6 of sample administration, and the weight and number of feces per subject were measured. In order to check the water content in feces, a change amount was calculated by comparing a weight immediately after collection with a weight after drying at 105° C. for 48 hours.

Water content (%)=[(weight before drying−weight after drying)/weight before drying]×100

[0134] The results were shown in Table 2 and FIG. 1.

TABLE-US-00002 TABLE 2 The number After administration of loperamide of Fecal weight Water content Group subjects (g) (%) Control group 1 (N) 5 0.373.sup.# 37.99* Control group 2 (N) 5 0.093 25.01 Control group 3 (PC) 5 0.163 30.03 Example 1 (M120) 5 0.195* 48.72.sup.# Example 2 (M500) 5 0.200* 44.64* Example 3 (M′120) 5 0.163 49.33.sup.#

[0135] 1) Table 2 shows the experimental results as means and standard errors. Significance was represented for each group as follows: * compared with P<0.05 C group, # compared with P<0.001 C group.

[0136] 2) N (Normal): normal control; C (Control): loperamide alone administered group; PC (Positive control): Psyllium husk 120 mg/kg; M120: loperamide+Tenebrionidae larva enzyme extract 120 mg/kg; 2500: loperamide+Tenebrionidae larva enzyme extract 500 mg/kg; M′120: loperamide+Tenebrionidae larva ethanol extract 120 mg/kg

[0137] As shown in Table 2 and FIG. 1, the fecal weight showed a significant decrease in group C causing constipation, compared to group N without causing constipation, but in all of groups M120, M500, and M′120 administered with the Tenebrionidae larva extract, the fecal weight was increased as compared to group C, and larger than that in group PC, administered with Psyllium husk.

[0138] In addition, the water content in feces showed a significant decrease in group C causing constipation, compared to group N without causing constipation, but in all of groups M120, M500, and M′120 administered with the Tenebrionidae larva extract, the water content in feces was significantly high as compared with group C, and higher than that of the normal control group.

[0139] As a result, it was confirmed that the Tenebrionidae larva extract prevented the decrease in fecal weight and improved the bowel movement function by increasing the water content in feces, so that it was confirmed that an effect of preventing or improving constipation was excellent.

Experiment Example 3: Changes in Digestive Tract Mobility Rate by Tenebrionidae Larva Extract

[0140] In order to observe an effect of a Tenebrionidae larva extract on a gastrointestinal transport capacity, the digestive tract mobility rate was evaluated. The digestive tract mobility rate may be evaluated by measuring the length of movement of an administered indicator material out of the entire length of the small intestine at a certain time, and in this experiment, loperamide, which exhibited a gastrointestinal transport inhibition effect, was used as a drug for causing constipation, and phenol red was used as an indicator material.

[0141] Specifically, 5 mg/kg of loperamide was orally administered after 1 hour of the administration of the Tenebrionidae larva extract. After 30 minutes of loperamide administration, 0.5% phenol red (in 1.5% methylcellulose) serving as an indicator was orally administered, and autopsy was performed after 20 minutes to measure the total intestinal length and a movement distance of phenol red. The measurement results were analyzed using the following equation, which was calculated as a percentage of the movement distance of phenol red with respect to the total intestinal length. The analysis results were shown in Table 3 and FIG. 2.

Digestive tract mobility rate (%)=(movement distance of Phenol red/total intestinal length)×100

TABLE-US-00003 TABLE 3 The number of Digestive tract Group subjects mobility rate (%) Control group 1 (N) 5 49.0.sup.# Control group 2 (N) 5 21.1 Control group 3 (PC) 5 28.8* Example 1 (M120) 5 28.6* Example 2 (M500) 5 34.5.sup.# Example 3 (M′120) 5 35.8.sup.#

[0142] 1) Table 3 shows the experimental results as means and standard errors. Significance was represented for each group as follows: * compared with P<0.05 C group, # compared with P<0.001 C group.

[0143] N (Normal): normal control; C (Control): loperamide alone administered group; PC (Positive control): Psyllium husk 120 mg/kg; M120: loperamide+Tenebrionidae larva enzyme extract 120 mg/kg; M500: loperamide+Tenebrionidae larva enzyme extract 500 mg/kg; M′120: loperamide+Tenebrionidae larva ethanol extract 120 mg/kg

[0144] As shown in Table 3 and FIG. 2, the digestive tract mobility rate was significantly reduced in group C causing constipation, compared to group N, whereas it was confirmed that in the Tenebrionidae larva extract administered group, the digestive tract mobility rate was significantly increased compared to group C (p<0.05, p<0.001). Furthermore, it was shown that in the Tenebrionidae larva extract administered group, the digestive tract mobility rate was higher than that of group PC administered with the Psyllium husk extract, and the digestive tract mobility rate was particularly high in the M′120 group.

[0145] From this, it was confirmed that the Tenebrionidae larva extract activated the digestive movement of the intestine, and as a result, it was confirmed that the Tenebrionidae larva extract can be effectively applied to the prevention or improvement of constipation.

Experiment Example 4: Confirmation of Structural Changes in Colon Tissue by Tenebrionidae Larva Extract

[0146] To determine an effect of the Tenebrionidae larva extract on the colon tissue, the colon tissue was stained with Hematozyln & Eosin (H&E).

[0147] Specifically, the colon tissue extracted from each experimental group was immobilized in 4% paraformaldehyde immediately after extraction, washed with water, dehydrated, transparent, and penetrated, and then embedded with paraffin. The prepared paraffin block was micro-sectioned and sliced, and then stained with H&E staining. The stained slides were photographed after observation with an optical microscope (Nikon, E600, Japan), and the results were shown in FIG. 3.

[0148] As illustrated in FIG. 3, it was observed that the length and area of the mucous membrane were significantly reduced in group C causing constipation, compared to non-treated group N, and it was confirmed that in the Tenebrionidae larva extract administered group, the length and area of the mucous membrane were increased compared to group C.

[0149] From this, it was confirmed that the Tenebrionidae larva extract could help the digestive movement of the intestine, and as a result, it was confirmed that the Tenebrionidae larva extract can be effectively applied to the prevention or improvement of constipation.

Example 5: Confirmation of Effect of Inhibiting Lipid Accumulation in Body by Tenebrionidae Larva Extract

[0150] In order to confirm an effect of the Tenebrionidae larva extract on lipid accumulation in the body, biochemical analysis in serum was performed.

[0151] Specifically, the experimental animals were fasted for 12 hours at the time when sample feeding was completed, and then the fasting blood was collected from the tail, and total cholesterol in the serum was measured using a blood analysis device. The measurement results were shown in Table 4.

TABLE-US-00004 TABLE 4 The number of Total cholesterol Group subjects (mg/dL) Control group 1 (N) 5 146.99 Control group 2 (N) 5 141.12 Control group 3 (PC) 5 136.72 Example 1 (M120) 5 184.44 Example 2 (M500) 5 179.14 Example 3 (M′120) 5 123.95

[0152] 1) N (Normal): normal control; C (Control): loperamide alone administered group; PC (Positive control): Psyllium husk 120 mg/kg administered group; M120: loperamide+Tenebrionidae larva enzyme extract 120 mg/kg; M500: loperamide Tenebrionidae larva enzyme extract 500 mg/kg; M′120: loperamide+Tenebrionidae larva ethanol extract 120 mg/kg

[0153] As shown in Table 4, it was confirmed that total cholesterol levels in the serum of the control group and Examples were 146.99 mg/dL and 141.12 mg/dL in non-treated group N and group C causing constipation, respectively, but the cholesterol level was reduced in the group M′120 administered with the Tenebrionidae larva ethanol extract after causing constipation.

[0154] Considering the above results, the Tenebrionidae larva extract showed significant increases in fecal weight, water content in feces, digestive tract mobility rate, and length and area of the mucous membrane compared to a loperamide-administered control group, so that the very excellent effect of improving the bowel movement function and improving intestinal motility was exhibited.