TW202220682A - Symbiotic composition with oligosaccharides ofgracilaria coronopifolia - Google Patents

Abstract

This invention discloses a symbiotic composition with oligosaccharides of Gracilaria coronopifolia, wherein the symbiotic composition comprises Lactobacillusor Bifidobacteriumas probitics; and water extract of Gracilaria coronopifoliaas prebiotic. The symbiotic composition of the invention may promote intestinal health.

Description

Synbiotics composition of oligosaccharides from corymbria

The present invention relates to a synbiotic composition, in particular, to a synbiotic composition containing corymbria oligosaccharide.

Diet is an important factor affecting the microbial composition of the gastrointestinal tract. The microorganisms in the gastrointestinal tract are closely related to the health of the human body. The dietary pattern affects the distribution, metabolic activity and colonization of the intestinal flora. Eating a balanced diet and consuming high-fiber foods or lactic acid bacteria products will help increase the number of good bacteria in the gut and reduce the number of bad bacteria.

Probiotics are defined as live bacteria that are believed to be beneficial to the host and promote the health of the host. Clinical evidence shows that Lactobacillus and Bifidobacterium have health-promoting effects and are often used in fermented milk products. Prebiotics are substances that can be used by probiotics to promote the balance of intestinal flora, such as oligosaccharides, dietary fibers, etc. Symbiotic is a compound substance of probiotics and probiotics, which can not only exert the physiological activity of probiotics, but also selectively increase the number of probiotics, increase the time that probiotics act in the intestine, and improve the effect of probiotics. Adaptability, the mutual dependence of the two. As of July 2016, there are as many as 75 items in my country's health food products aimed at improving gastrointestinal function, most of which are probiotics or prebiotics.

Seaweed is rich in dietary fiber, protein, minerals (potassium, calcium, magnesium and manganese) and omega-3 polyunsaturated fatty acids, which are extremely valuable in the development of medical and health care products. However, there are few combinations of seaweed and probiotics. Applications.

In view of the foregoing, the inventors of the present invention have painstakingly researched, considered and designed a synergetic element containing coriander mustard oligosaccharide, in order to improve the deficiencies in the prior art, thereby promoting the implementation and utilization in the industry.

In view of the above-mentioned problems in the prior art, the purpose of the present invention is to provide a synbiotic composition of oligosaccharides of Corymbria corymbria, which is used to solve the deficiencies of the prior art.

Based on the above purpose, the present invention provides a synbiotic composition of coriander mustard oligosaccharide, which comprises probiotics and probiotics. The prebiotic contains coriander mustard water extract. Probiotics include lactobacilli, bifidobacteria, or a combination thereof.

Preferably, the preparation method of the mustard coriander water extract comprises the following steps: mixing the mustard coriander powder with hot water in a ratio of 50:50, and heating the mustard coriander powder to a temperature of 60°C for processing After 6 hours, centrifugation was performed again to obtain the supernatant, and the supernatant was freeze-dried overnight to obtain the powder of the water extract.

Preferably, the synbiotic composition comprises probiotics in powder form and probiotics in powder form prepared from water extracts, and the probiotics and probiotics are mixed in a ratio of 2:8 to 8:2 .

Preferably, the probiotics are selected from at least one of Lactobacillus and Bifidobacterium.

Preferably, the probiotics comprise strains with BCRC numbers BCRC 11844, BCRC 14602, BCRC 11847, BCRC 10695 or BCRC 16053.

Preferably, the synbiotic composition of the present invention further comprises skim milk powder as a protective agent.

Preferably, the added amount of the protective agent can be 10% to 20% of the total weight of the probiotics and prebiotics.

Continuing from the above, according to the synergistic element of the oligosaccharide of A. corymbria of the present invention, the cell damage and inflammatory reaction caused by reactive oxygen species can be reduced, and the gastrointestinal diseases caused by improper diet or human aging can be improved. Intestinal health care and prevention of intestinal inflammation, to achieve the effect of promoting intestinal health.

In order to make the above-mentioned purpose, technical features, and gain after actual implementation more obvious and easy to understand, a more detailed description will be given below with the aid of a preferred embodiment example and corresponding related drawings.

The present invention provides a synbiotic composition of oligosaccharides of Corymbria coriander, which comprises probiotics and probiotics. The probiotic used in the present invention is the water extract of A. corymbria, which can be the water extract obtained by extracting A. corymbria with hot water. The probiotic can be selected from at least one of Lactobacillus and Bifidobacterium in the intestinal tract.

The form of the synbiotic composition can be a powder or a liquid additive. The synbiotic composition stored in the powder form is described in the following embodiments of the present invention, but not limited thereto. When implementing or using the synbiotic composition of the present invention, it can be adjusted into powder, granule, ingot, crystal ball, liquid and milk form according to the commodity. In another embodiment, the synbiotic composition may include skimmed milk powder as a protective agent, the content of which is 100% based on the total weight of probiotics and prebiotics, and 10% to 20% of skim milk is added. pink.

Taking a powdered synbiotic composition as an example, it may include powdered probiotics and powdered probiotics prepared from water extracts. Mix in a ratio of 8:2.

Experimental Materials

1. Seaweed Extract

The seaweed extract used in the present invention is Gracilaria coronopifolia collected in Taiwan, and is used after pre-processing such as washing, freeze drying and pulverization. Preparation of the extract: In the present invention, the powder and hot water are extracted by heating to a temperature of 60° C. for 6 hours according to the weight ratio of 50:50 (powder: hot water), and then centrifuged to obtain the filtrate as the heat. The water-soluble extract was vacuum freeze-dried and the extract was stored.

2. Strains

The used bifidobacteria include 3 strains including Bifidobacterium bifidums BCRC 11844, Bifidobacterium longum subsp.infantis BCRC 14602, Bifidobacterium longum subsp.longum BCRC 11847, and 2 strains including Lactobacillus acidophilus BCRC 10695 and Lactobacillus delbrueckii subsp.bulgaricus BCRC 16053. Purchased from the Biological Resource Conservation and Research Center, Hsinchu Food Industry Development Institute.

After inoculation 2-3 times before the test, inoculate 1% of the strain to 10 ml of the appropriate medium for each strain, inoculated in fortified Clostridium medium (RCM), Lactobacillus medium (MRS) or MRS+0.05% cysteine, and Cultivated at 37°C until the absorbance of the bacterial solution (wavelength 610nm) reached 0.8 (1.0× ^{10 8} CFU/ml), which was used as a test.

3. Seaweed Synbiotics

The seaweed synbiotics used in this experiment are the freeze-dried powder of coriander mustard extract and 5 strains mixed in different weight ratios to form GS1 (strain:extract=30:70), GS2 (strain) : extract = 50: 50), GS3 (strain: extract = 70: 30), the weight of each strain in the mixed strain is the same.

4. Cell Lines

A total of 2 cell lines were used in this experiment, human intestinal cell C2BBel BCRC 60182 (human colon adenocarcinoma, clone of Caco-2) and human monocyte BCRC 60430 (human acute monocytic leukemia, THP-1), which were purchased from Consortium Foods Industrial Development Institute Biological Resources Conservation and Research Center.

experimental method

1. Cell Culture

Caco-2 cells were cultured in DMEM (Dulbecco's modified eagle's medium) medium (containing 10% fetal bovine serum, Fetal bovine serum, FBS) at 37 °C and 5% CO2 concentration. Subculture.

THP-1 cells were cultured in RPMI (Roswell Park Memorial Institute) 1640 medium (containing 10% FBS) at 37°C under 5% CO2 concentration. Dilute cells directly into fresh medium before aliquoting into new culture flasks. Or centrifuge the cells containing the medium at low speed (1000rpm, 5 minutes), remove the supernatant, add fresh medium and mix evenly, transfer to a new culture flask, wait for about 2-3 days for cells to become full, and then continue. Generation cultivation.

2. Co-culture of Caco-2 and THP-1

Referring to the literature published by Takisawa et al. in 2009 (Anti-Inflammatory effect of Bifidobacterium longum on macrophage-like THP-1 cells via epithelial cell Caco-2), a transwell filter was used for co-culture. Caco-2 cells were cultured to form a monolayer cell membrane, the lower layer was THP-1 cells, and the lower layer was stimulated with 10ng/ml LPS and 10ng/ml TNF-α, co-cultured for 24 hours, and the lower cell culture medium was collected for subsequent experimental analysis.

3. Cell viability assay (MTT assay)

After the cells were collected by centrifugation at 1500 rpm for 5 minutes, the cell density was calculated by a cell counter (Sequoia-Turner Cell-dyn 300), and then the cells were adjusted to 2×10 ⁵ cells/ml according to this value, and added to 96 wells at 100 μl/well. Place the culture dish in a 37°C, 5% CO ₂ cell incubator for 24 hours, extract the original culture medium, add 100 μl of samples of different concentrations (0.1-1000 μg/ml), and incubate at 37°C. , 5% CO ₂ for 24 hours, add 100 μl of MTT (3-4,5-Dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide) to each well, place at 37°C to react for 4 hours, and finally add 100 μl of DMSO was used to dissolve the cell membrane and mixed evenly to ensure that the blue-violet crystals were completely dissolved. After standing for 40 minutes, the absorbance at a wavelength of 570 nm was read with an ELISA reader. The culture medium was used to replace the sample as the control group, which was regarded as 100%, and the blank group was the culture medium without cells and samples.

Cell viability (%)=[(absorbance value of sample group-absorbance value of blank group)/absorbance value of control group]×100%.

4. Monolayer Cell Transmembrane Resistance Value (TEER) Determination

Caco-2 cells were seeded in Millicell insert culture dishes (PCF membrane, 0.4 μm pore size), the growth of monolayer cell membrane was observed, and the TEER value was measured at different times with a Millipore ERS resistance meter, and monitored for 21 days. When measuring TEER, a pair of electrodes are placed on the top and base of the cell monolayer respectively, and the actual resistance value is measured with a resistance meter, which is calculated according to the following formula:

TEER=(R _total -R _blank ) × A(Ω×cm ² )

where R _total is the measured resistance value, R _blank is the blank film resistance value, and A is the film area.

5. Determination of cellular reactive oxygen species (ROS) content

Take 5×10 ⁵ cells/ml and seed them in a 12-well plate containing 2% serum, add samples and H ₂ O ₂ , control the concentration of H ₂ O ₂ in the medium to 400 μM, at 37 ^o C, 5 Incubate in a constant temperature incubator with % CO ₂ for 12 hours, and 30 minutes before the end of the reaction, then add 20 μM of the fluorescent dye 2',7'-dichlorofluorescin diacetate (DCFH-DA) to the culture medium to react for 30 minutes. The cells were then centrifuged at 400 g for 10 minutes and the supernatant was removed, washed twice with PBS, 0.5 ml of PBS was added to suspend the cells, and transferred to a 96-well black dish, and the cells were analyzed by a fluorescence analyzer (Hitachi F-4500, Hitachi Co. . Ltd., Tokyo, Japan) to detect fluorescent reactions (Excitation 485 nm, Emission 527 nm).

6. Cytokine (IL-8) Analysis

The concentration of cultured Caco-2 cells was adjusted to 3×10 ⁵ cells/ml, cultured in a 6-well dish for 21 days, and then differentiated with PMA (phorbol-12-myristate-13-acetate) 200 nM for 4 days. The THP-1 cells were co-cultured. Inject 1.5 ml samples of different concentrations (0.1-1000 μg/ml) into the inner chamber, add 2 ml RPMI 1640 medium to the outer chamber, and place it in a cell incubator at 37°C, 5% CO ₂ for 3 hours, then add 100 ng/ml ml TNF-α into the outer chamber, activated for 24 hours. The suspension in the well plate was collected and stored at -20°C for use in the determination of the cytokines contained in it. The assay kit was used for the detection of cytokines human IL-8 ELISA (431505).

A 96-well test plate with monoclonal antibody attached was prepared, and after diluting the capture antibody with PBS, 100 μl was added to each well, and it was placed at 4°C overnight. Wash buffer) for 4 times, add 200 μl of block buffer to each well, and react at room temperature for 1 hour, aspirate the solution in the well plate, rinse 4 times with washing buffer, and the test plate is completed. . Add 100 μl of culture medium to the test plate, react at room temperature for 2 hours, aspirate the medium in the well plate after 2 hours, wash 4 times with washing buffer, add 100 μl of detection antibody, and incubate at room temperature After 1 hour of action, the solution in each well plate was aspirated, washed 4 times with washing buffer, added 100 μl of horseradish peroxidase-labeled streptavidin (Streptavidin-HRP), and reacted at room temperature for 30 After 20 minutes, aspirate the solution in each well plate, rinse with washing buffer for 5 times, add 100 μl of substrate solution, react at room temperature for 20 minutes in the dark, and stop the reaction by adding 100 μl after 20 minutes. solution (Stop solution), and the absorbance value of each well plate at a wavelength of 450 nm was measured using an ELISA reader.

result

1. Preparation and storage test of seaweed synbiotics

The coriander mustard extract is used as a seaweed probiotic, mixed with 5 strains of Lactobacillus and Bifidobacterium, and 15% skimmed milk powder is added as a protective agent. It is generally recognized that the bacterial count of probiotic products containing live bacteria needs to be maintained above 10 ⁶ cfu/g to exert its activity, so 10 ⁶ cfu/g is used as the screening condition for the storage period. In order to understand the storage stability of the prepared seaweed synbiotics (GS1, GS2, GS3), tests were conducted at different temperatures (4°C, -20°C). The seaweed synbiotics were stored at 4°C for 60 days (Figure 1 (a) part), the bacterial count was still maintained at 10 ⁹ cfu/g, and stored at -20°C for 60 days (part (b) of Figure 1), the bacterial count of GS1 and GS3 remained at 10 ⁶ -10 ⁸ cfu/g, It shows that it can delay the reduction of the bacterial count of seaweed and synbiotics at low temperature, and the low temperature storage is stable.

2. Discussion on the survival rate of Caco-2 cells by seaweed synbiotics

In this study, an in vitro evaluation model was used to explore the use of seaweed synbiotics as a material for preventing intestinal inflammation, and intestinal cell lines were used as the research object. In order to understand the effect of the test material on the human intestinal cell line Caco-2, the cell viability after 24 hours of treatment with compound probiotics and coriander mustard extract. At the concentration of 0.1-1000 μg/ml of coriander mustard extract, the cell viability was 99-110%, 104-112%, 93-106%, respectively. The results showed that at this concentration, the compound probiotics and the umbrella Venus mustard extract does not cause harm to intestinal cells. In addition, the cytotoxicity of seaweed synbiotics on Caco-2 was tested, and the results showed that the cell viability of GS1 to Caco-2 was 102%-115%, and the cell viability of GS2 to Caco-2 at a concentration of 0.001-1 mg/ml The cell survival rate of GS3 to Caco-2 is 101%-117%, that is, the three groups of formulations are not toxic to Caco-2 cell line.

3. Establishment of a co-culture model of Caco-2 cells and THP-1 macrophages

Using Caco-2 cells as the evaluation model, the degree of change in transepithelial electrical resistance (TEER) was detected, and the value should be greater than 600Ωcm ² to confirm the formation of monolayer. The TEER value after 14 days of culture was: 685Ωcm ² (as shown in Figure 2), and then observed under a microscope, the required incubation time to form a complete monolayer was 21 days and 1053Ωcm ² (as shown in Figure 2). THP-1 cells are cell lines established from monocytes in the blood of leukemia patients. They can secrete inflammatory factors and transform into macrophages after moderate induction. They are good substitute cells for the function of human monocytes and macrophages in vitro. In this experiment, THP-1 human monocytes were cultured to establish an in vitro model of their differentiation into macrophage-like cells. THP-1 cells were differentiated with 200 nM PMA (Phorbol-12-myristate-13-acetate). After 4 days, the cell morphology had transformed into a macrophage-like pattern. The two cell lines were co-cultured on an insert cell culture dish, with Caco-2 cells in the upper layer and THP-1 macrophages in the lower layer.

4. The effect of seaweed synbiotics on intestinal cell monolayer

Under the co-culture of Caco-2 cells and THP-1 macrophages, seaweed synbiotics were added to the upper cells, and samples were taken from the lower cell culture medium at regular intervals to observe the permeability of the samples to intestinal cells, and Taking FD-4 (fluorescein isothiocyanate dextran tracer 4 kDa, MW4400) as the positive control group, the penetration rate was calculated, and the TEER before and after the experiment was detected. The seaweed synbiotics were treated to co-culture cells at the concentration of 0.001-0.1 mg/ml. The results are shown in Figure 3. The transmembrane resistance value relative to FD-4 (100%), treated with 0.1 mg/mL GS1 synbiotic, was 86% at the beginning of the experiment and increased to 95% after 180 minutes , in addition, after 180 minutes of treatment 0.001 mg/mL, 0.005 mg/mL and 0.05 mg/mL were 105%, 103% and 101%, respectively. The TEER value of seaweed synbiotics can still maintain more than 80% during the action time of 3 hours, that is, seaweed synbiotics will not cause damage to the monolayer membrane of intestinal cells.

5. To investigate whether cells have protective effect under oxidative stress with seaweed synbiotics

Different concentrations (0.125-2 mM) of H ₂ O ₂ were used to induce THP-1 cells, and the cell viability was investigated after 24 hours of co-culture. As shown in Table 1, the concentration of H ₂ O ₂ was negatively correlated with the viability. The dose-dependence, in which the cell viability after 1 mM H ₂ O ₂ treatment was 67.4%, indicated that H ₂ O ₂ at this concentration would produce oxidative stress on cells, but would not kill cells. 1-20 ng/ml of TNF-α was added to the cells, and the survival rate was measured after co-cultivation for 24 hours. Table 2 shows that the survival rate was still 75.9% at a concentration of 20 ng/ml, which also had a concentration-dependent negative correlation. sex. Therefore, 1 mM H ₂ O ₂ and 20 ng/ml TNF-α were used as subsequent concentrations to induce oxidative stress.

Table I _{H2O2 concentration} 0.125mM 0.25mM 0.5mM 1 mM 2 mM Survival rate (%) 95.49 83.298 79.035 67.415 35.941 *100% with no treatment as the control group

Table II TNF-α concentration 1 ng/ml 2ng/ml 5ng/ml 10ng/ml 20ng/ml Survival rate (%) 98.05 92.867 88.398 82.793 75.891 *100% with no treatment as the control group

The seaweed synbiotics were co-cultured with THP-1 cells, and ₁ mM H ₂ O ₂ and ₂₀ ng/ml TNF-α were added to induce the formation of oxidative stress, and the cell viability was tested. Under induction, the cell viability decreased to 70.7%. After adding seaweed synbiotics, the cell viability was improved. The GS1 group increased to 80.4-100.3%, the GS2 group increased to 82.1-105.5%, and the GS3 group increased to 70.7- 81.20%; it can be seen from Table 4 that under the induction of THF-α, the cell viability decreased to 74.3%, and after treatment with GS1 (0.005-0.5 mg/ml), the cell viability increased to 79.68-92.2%, and the GS2 group (0.001- 0.5 mg/ml) to 75.7-94.2%, and GS3 group (0.005-0.1 mg/ml) to 76.5-82.4%. It can be seen that seaweed synbiotics can reduce cell damage caused by oxidative stress (H ₂ O ₂ and TNF-α), and improve cell survival rate.

Table 3 H ₂ O ₂ 1mM Seaweed Synbiotics 0.001 mg/ml 0.005 mg/ml 0.01 mg/ml 0.05 mg/ml 0.1 mg/ml 0.5 mg/ml GS1 70.66 90.15 95.81 86.99 85.19 100.34 80.38 GS2 70.66 93.74 90.54 95.29 105.46 103.88 82.12 GS3 70.66 81.20 76.50 71.74 70.68 73.17 75.35 Unit: cell proliferation rate (%), untreated as the control group, 100%

Table 4 TNF-α 2 ng/ml Seaweed Synbiotics 0.001 mg/ml 0.005 mg/ml 0.01 mg/ml 0.05 mg/ml 0.1 mg/ml 0.5 mg/ml GS1 74.37 72.91 79.68 75.79 81.96 89.89 92.21 GS2 74.37 75.69 87.33 83.32 94.16 80.64 86.37 GS3 74.37 66.99 82.36 76.47 77.94 81.22 65.61 Unit: cell proliferation rate (%), untreated as the control group, 100%

1 mM H ₂ O ₂ was used to induce cellular oxidative stress to form ROS, and the experimental concentrations of 0.1 mg/ml GS1, 0.05 mg/ml GS2 and 0.005 mg/ml GS3 were used to investigate whether seaweed synesthesia could reduce THP-1 cells. oxidative stress. The results showed that the amount of ROS production increased to 151.5% (100% in the unexcited group) after stimulation with 1 mM H ₂ O ₂ , while the amount of ROS generated after treatment with seaweed synbiotics (GS1, GS2, GS3) They were reduced to 101.751%, 101.363% and 109.582%, respectively, that is, the algae synbiotics could slow down the production of ROS induced by H ₂ O ₂ and reduce the oxidative stress of THP-1 cells.

On the Caco-2 cell platform, because the concentration of 1 mM H ₂ O ₂ will cause Caco-2 cells to suspend, so reducing the concentration of induction to 0.5 mM, the survival rate is 94.1% (as shown in Table 5).

Table 5 _{H2O2 concentration} 0.5mM 1mM 2 mM 4mM 5mM Survival rate (%) 94.054 84.933 48.740 31.788 16.023 *100% with no treatment as the control group

As shown in Table 6, after excitation with 0.5 mM H ₂ O ₂ , the ROS production increased to 105.4%, and after treatment with seaweed synbiotics (GS1, GS2, GS3), the ROS production decreased to 63.3-84.3%, 48.8-81.0% and 56.2-74.1%.

Table 6 H ₂ O ₂ 0.5 mM Seaweed Synbiotics 0.001 mg/ml 0.005 mg/ml 0.01 mg/ml 0.05 mg/ml GS1 105.4 84.3 81.1 67.3 63.3 GS2 105.4 62.8 89.5 81.0 48.8 GS3 105.4 74.1 66.0 56.2 68.6 Unit: fluorescence intensity (%), with untreated as the control group 100%

It can be seen from Table 7 that after stimulation with 20 ng/ml TNF-α, the production of ROS increased to 124.5% (100% in the non-excited group), while after treatment with seaweed synbiotics (GS1, GS2, GS3), ROS production increased to 124.5% (100% in the unexcited group). The production volume dropped to 93.0-106.6%, 60.6-81.1% and 57.7-91.3% respectively. That is, seaweed synbiotics can slow down the production of ROS induced by H ₂ O ₂ and TNF-α, reduce the oxidative stress of Caco-2 cells, and it is expected to prevent the inflammatory response caused by oxidative stress.

Table 7 TNF-α 20 ng/ml Seaweed Synbiotics 0.001 mg/ml 0.005 mg/ml 0.01 mg/ml 0.05 mg/ml GS1 124.5 95.2 103.6 93.0 106.6 GS2 124.5 60.6 76.2 81.1 72.8 GS3 124.5 78.2 57.7 63.0 91.3 Unit: fluorescence intensity (%), with untreated as the control group 100%

6. Evaluation of the adjustment of inflammatory factors by seaweed synbiotics

In order to explore whether the co-culture of Caco-2 and THP-1 is feasible, the reaction of 500 ng/ml LPS with THP-1 for 3 hours can induce THP-1 to secrete TNF-α, and cause surface damage of Caco-2 cells after 24 hours. TNF-α secretion was greatly increased. The production of TNF-α can induce Caco-2 to secrete interleukin-8 (IL-8), which shows that LPS and TNF-α induce intestinal cell inflammation, which increases the secretion of inflammatory factors. The experimental model is feasible. It can be seen from Table 8 that after the cells were induced to produce IL-8 (1.47 ng/ml) with 20 ng/ml TNF-α, the algae synbiotics were treated at a concentration of 0.001-0.05 mg/ml, in which GS1 was at 0.001 and 0.005 mg/ml. At the concentration of ml, the production of IL-8 was lower than that of IL-8 induced by TNF-α. At 0.001, 0.005 and 0.01 mg/ml of GS2, the production of IL-8 was higher than that of IL-8 induced by TNF-α. IL-8 production was reduced at 0.01 mg/ml by GS3. It was confirmed that seaweed synbiotics can reduce the production of IL-8 induced by TNF-α, reduce the inflammatory response of cells, and then prevent the occurrence of inflammatory response of intestinal cells.

Table 8 TNF-α 20 ng/ml Seaweed Synbiotics 0.001 mg/ml 0.005 mg/ml 0.01 mg/ml 0.05 mg/ml GS1 1.47 1.13 1.10 1.77 1.12 GS2 1.47 1.38 0.56 1.08 1.61 GS3 1.47 1.62 1.62 1.11 1.50 Unit: IL-8 content (ng/ml), the value of the untreated control group is 0.34 ng/ml

From the above results, it can be seen that the synbiotic composition containing coriander mustard not only does not affect the growth of intestinal cells, but can also reduce cell damage caused by oxidative stress, or reduce the production of inflammatory response, and improve cell growth. the survival rate. Therefore, it can improve gastrointestinal diseases caused by improper diet or human aging, and has intestinal health care and prevention of intestinal inflammation, so as to achieve the effect of promoting intestinal health.

The above description is exemplary only, not limiting. Any equivalent modifications or changes that do not depart from the spirit and scope of the present invention shall be included in the appended patent application scope.

none

Figure 1 shows the storage stability test results of seaweed synbiotics. Figure 2 shows the change in transmembrane resistance value of Caco-2 cells during 21 days of culture. Figure 3 is the TEER value of the co-culture system treated with seaweed synbiotics. Parts (a), (b), and (c) are the results of processing samples GS1, GS2, and GS3, respectively.

Claims (7)

A synbiotic composition of oligosaccharides of Coronocarpus corymbs, comprising probiotics and probiotics, wherein the probiotics comprise water extract of Gracilaria coronopifolia; the probiotics comprise Lactobacillus , Bifidobacterium or a combination thereof. The synbiotic composition as claimed in claim 1, wherein the preparation method of the water extract of Corymbria corymbria comprises the following steps: Mix coriander mustard powder with hot water in a ratio of 1:1, and The coriander mustard powder is heated to a temperature of 50-80° C., and then centrifuged and freeze-dried to obtain the coriander mustard water extract. The synbiotic composition according to claim 2, wherein the synbiotic composition comprises the probiotics in powder form and the probiotics in powder form, the probiotics and the probiotics are in a ratio of 2:8 Mix to a ratio of 8:2. The synbiotic composition according to claim 1, wherein the probiotic is selected from at least one of Lactobacillus and Bifidobacterium . The synbiotic composition according to claim 1, wherein the probiotics include BCRC 11844 ( Bifidobacterium bifidums ), BCRC 14602 ( Bifidobacterium longum subsp.infantis), BCRC 11847 ( Bifidobacterium longum subsp.longum), BCRC 10695 ( Lactobacillus acidophilus ) or BCRC 16053 ( Lactobacillus delbrueckii subsp. bulgaricus ) . The synbiotic composition according to claim 1, further comprising skimmed milk powder as a protective agent. The synbiotic composition according to claim 6, wherein the added amount of the protective agent is 10% to 20% of the total weight of the probiotic bacteria and the probiotics.

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