TWI787696B - Uses of a plant fermentation liquid in preparing composition for reducing lipid - Google Patents

Abstract

Uses of a plant fermentation liquid in preparing composition for reducing lipid. The plant fermentation liquid made from water extract of gardenia(Gardenia jasminoides ) and purslane(Portulaca oleracea )after first fermentation of Yeast and Lactobacillus and second fermentation of Acetic acid bacteria.

Description

Use of plant fermented liquid for preparing lipid-lowering composition

The present invention relates to a fermented liquid, in particular to the use of a fermented liquid of japonica and purslane for preparing a lipid-lowering composition.

Organic and natural diet concepts are gaining popularity as consumers are increasingly concerned about artificial substances or additives. Biotechnology companies and food companies are actively investing in research and development of related products related to natural products.

On the basis of various scientific verifications that are beneficial to human health, the analysis and efficacy evaluation of active ingredients for different plants has become a key item in product development. The product development project is to use natural plants to improve or enhance body functions, such as weight loss, whitening, gastrointestinal health care, ketogenic diet, etc.

Among them, low metabolism and high body fat are common health hazards of modern people, and low metabolism and high body fat may further lead to various diseases such as hyperlipidemia, fatty liver, arteriosclerosis and/or myocardial weakness.

In some embodiments, the plant fermentation broth is used to prepare a lipid-lowering composition. Among them, the plant fermentation liquid is obtained from water extracts of Gardenia jasminoides and Portulaca oleracea , which are firstly fermented by yeast and Lactobacillus, and then secondarily fermented by acetic acid bacteria.

In some embodiments, plant fermentation broth is used to lower cholesterol.

In some embodiments, plant fermentation broth is used to lower LDL.

In some embodiments, plant fermentation broth is used to increase HDL.

In some embodiments, plant ferments are used to reduce body fat.

In some embodiments, plant fermentation broth is used to reduce calcium ion deposition in smooth muscle cells.

In some embodiments, plant fermentation broth is used to enhance the activity of vascular cell granulocytes.

In some embodiments, plant fermentation broth is used to elevate extremity temperature.

In some embodiments, the effective dosage of the plant fermentation broth is 10 mL/day.

To sum up, according to any embodiment of the plant fermentation liquid, it can reduce cholesterol and low-density lipoprotein, and increase high-density lipoprotein, thereby reducing the blood fat content in the blood to achieve the purpose of lowering blood lipids. According to any embodiment of the plant fermentation liquid, it can greatly reduce the generation of calcium ion deposition in smooth muscle cells, thereby improving myocardial contractility and avoiding blood vessel blockage, thereby increasing metabolic rate to achieve lipid-lowering purposes. According to any embodiment of the plant fermentation liquid, it can increase the activity of vascular cell mitochondria, thereby improving the efficiency of blood vessels, and then increasing the metabolic rate to achieve the purpose of lowering lipids. According to any embodiment of the plant fermented liquid, it increases the body temperature, and then increases the metabolic rate to achieve the purpose of reducing fat. Moreover, according to any embodiment of the plant fermented liquid, the lipid-lowering purpose can be effectively achieved by taking 10 mL of the plant fermented liquid every day.

In some embodiments, the plant fermentation liquid is obtained by primary fermentation of the water extract of kale seed and purslane through yeast ( Yeast ) and Lactobacillus ( Lactobacillus ) and secondary fermentation with acetic acid bacteria ( Acetobacter aceti ) be made of. In some embodiments, the water extract is prepared from the raw materials of japonica oleracea, purslane raw materials and water. In some embodiments, the water extract is prepared from the raw materials of kale seed, purslane raw material, water and glucose.

In some embodiments, the plant of Gardenia jasminoides is an evergreen shrub of the Rubiaceae ( Rubiaceae ) genus ( Gardenia ), also known as Mudan and fresh branch. In some embodiments, the raw material of japonica is japonica fruit. In some embodiments, the raw material of japonica japonica is shelled fruit of japonica japonica. In some embodiments, the raw material of japonica is dried japonica pulp and seeds. For example, as the raw material of japonica, shelled fruit of japonica procured from China (Lu Sejong) can be used.

In some embodiments, the plant of purslane ( Portulaca oleracea ) is a herbaceous plant belonging to the genus Portulaca of the family Portulacaceae , also known as horse lettuce, portulaca oleracea, purslane and five elements grass. In some embodiments, the raw material of purslane is the whole herb of purslane. In some embodiments, the raw material of purslane is the stem and leaves of purslane. In some embodiments, the raw material of purslane is purslane leaf. For example, dried purslane leaves purchased from China (Lu Sejong) can be used as the raw material of purslane.

In some embodiments, the water extract is prepared from the raw materials of kale seed, purslane raw material and water in a weight ratio of 1-3:0.5-1.5:75-85. In some embodiments, the water extract is prepared from the raw materials of kale seed, purslane raw material and water in a weight ratio of 2:1:80. In some embodiments, the water extraction solution is to mix the raw materials of kale seed, purslane raw material and water according to the ratio of 2:1:80, and then carry out high-temperature extraction for 60 minutes to 70 minutes (such as, 90±5 ℃) made. In some embodiments, the water extract is prepared by mixing the raw materials of kale seed, purslane raw material and water according to the ratio of 2:1:80 and then performing high-temperature extraction (eg, 95° C.) for 60 minutes. . In some embodiments, the aqueous extract has a Brix degree (Brix°) greater than or equal to 8. In some embodiments, the water extract is prepared from the raw materials of kale seed, purslane raw material, water and glucose. Wherein, the weight ratio of the raw material of kale seed, the raw material of purslane and water is 2:1:80, and the content of glucose is 10% (V/ V). In some embodiments, the water extraction solution is mixed with the raw materials of kale seed, purslane raw material and water according to the ratio of 2:1:80, then adding 10% glucose, and then performing high-temperature extraction for 1 hour (eg, 95°C).

In some embodiments, the water extract is directly added to the strains for fermentation without filtering out the solids inside (that is, the raw material of kale seed / or the raw material of purslane), so as to use the strains to further extract the activity in the solids Components, and then get the plant fermentation liquid.

In some embodiments, after high-temperature leaching, the temperature of the water extract is lowered for use in subsequent fermentation procedures. In some embodiments, after high-temperature leaching, the water extract solution is cooled to 35° C.±2 for use in subsequent fermentation procedures.

In some embodiments, after inoculating the strain with the water extract, a fermentation process is performed to obtain a plant fermentation liquid, and the fermentation process is divided into a primary fermentation process and a secondary fermentation process. First, the initial fermentation procedure is to add 0.05wt%~0.15wt% of yeast and 0.025wt%~0.1wt% of lactic acid bacteria to the water extract and leave it to ferment at room temperature for 2 days to 5 days to form the initial fermentation liquid. In some embodiments, the initial fermentation procedure is to add 0.1wt% of yeast and 0.05wt% of lactic acid bacteria to the water extract and leave it to ferment at 25°C to 28°C for 3 days to form the initial fermentation liquid.

In some embodiments, the yeast may be Saccharomyces cerevisiae . For example, the lactic acid bacteria can be the strain Saccharomyces cerevisiae deposited at the Bioresource Conservation and Research Center (BCRC) of the Food Industry Development Institute with the deposit number BCRC 20271 (and the international deposit number ATCC26602) or other commercially available brewer's yeast.

In some embodiments, the lactic acid bacteria may be Lactobacillus plantarum. For example, the lactic acid bacteria can be the TCI028 strain deposited in the Bioresources Conservation and Research Center of the Food Industry Development Institute and deposited in the number BCRC 910805 strain, and this strain is also deposited in the German Collection of Microorganisms and Cell Cultures , DSMZ) and its international deposit number is DSM33108.

Next, the primary fermentation broth obtained in the primary fermentation procedure is subjected to a secondary fermentation procedure. In some embodiments, the secondary fermentation procedure is to add 5wt% acetic acid bacteria into the initial fermentation broth and let it stand still and detect that the Brix is less than 4, and the fermentation is completed when the acid-base value (pH value) is 4.5±0.5, The plant fermentation stock solution is formed. In some embodiments, the secondary fermentation process is to add 5wt% acetic acid bacteria to the initial fermentation liquid and leave it to ferment at room temperature for 3 to 6 days to form a plant fermentation stock solution. In some embodiments, the secondary fermentation process is to Add 5wt% acetic acid bacteria into the solution and let it stand for fermentation at 25°C to 28°C for 4 days to form a plant fermentation stock solution. In some embodiments, acetic acid bacteria strains with accession number BCRC11688 (international deposit ATCC15973) are used.

In some embodiments, the plant fermentation liquid obtained from the fermentation procedure is the plant fermentation liquid. In some other embodiments, the plant fermentation stock solution obtained after the fermentation procedure can be subjected to at least one of the following reprocessing procedures: filtration procedure, vacuum concentration procedure, seasoning procedure, filling procedure and sterilization procedure, so as to form a plant fermentation broth. Here, the filtration procedure, decompression concentration procedure, seasoning procedure, filling procedure or sterilization procedure are used to prolong the shelf life and taste of the plant fermented liquid and avoid deterioration.

In some embodiments, the filtering procedure is to filter the plant fermentation liquid through a 400-mesh filter to form a plant fermentation liquid. Here, the solids are removed through a filter of appropriate mesh.

In some embodiments, the vacuum concentration procedure is to concentrate the plant fermentation liquid at 55°C to 65°C under reduced pressure to form a plant fermentation liquid. For example, the temperature set value for concentration under reduced pressure is 60°C. In some embodiments, the set pressure of the reduced-pressure concentration is 150 bar (Bar). Here, the alcohol content in the plant fermentation broth can be removed by concentration under reduced pressure, and the storage volume of the plant fermentation broth can be reduced.

In some embodiments, the seasoning procedure is to add oligosaccharides to the plant fermentation liquid to form a plant fermentation liquid. Herein, oligosaccharides refer to oligosaccharides formed by polymerization of 3-10 monosaccharide molecules. In some embodiments, the oligosaccharides can be fructooligosaccharides, galactooligosaccharides, xylooligosaccharides, isomaltooligosaccharides, and the like. In one embodiment, 40% to 70% of the oligosaccharides relative to the plant fermentation stock solution are added. In one embodiment, 60% oligosaccharides relative to the plant fermentation stock solution are added. Here, adding oligosaccharides is to adjust the osmotic pressure of the plant fermentation broth, reduce the water activity in the plant fermentation broth and avoid the growth of other microorganisms and affect the quality of the plant fermentation broth.

In some embodiments, the filling procedure is to divide the plant fermentation broth/or plant fermentation stock solution into storage containers of appropriate size. In some embodiments, the filling procedure is to divide the plant fermentation broth/or the plant fermentation stock solution into 10 mL storage containers.

In some embodiments, the sterilization procedure is to heat the plant fermentation broth/or the plant fermentation stock solution to 95° C.±5 for at least 60 minutes.

In some embodiments, plant fermentation broth is used to lower cholesterol. In some embodiments, plant fermentation broth is used to lower LDL. In some embodiments, plant fermentation broth is used to increase HDL. In some embodiments, plant ferments are used to reduce body fat.

The medical field usually uses the detection of body fat, high total cholesterol (Cholesterol) in human blood, high low-density lipoprotein (LDL) or low high-density lipoprotein (HDL) as indicators of health status, and total cholesterol ( Cholesterol), high low-density lipoprotein (LDL), or low high-density lipoprotein are known causes of hyperlipidemia.

In some embodiments, plant fermentation broth is used to reduce calcium ion deposition in smooth muscle cells. In some embodiments, plant fermentation broth is used to enhance the activity of vascular cell granulocytes. In some embodiments, plant fermentation broth is used to elevate extremity temperature.

In some embodiments, the effective dosage of the plant fermentation broth is 10 mL/day.

To sum up, according to any embodiment of the plant fermentation liquid, it can reduce cholesterol and low-density lipoprotein, and increase high-density lipoprotein, thereby reducing the blood fat content in the blood to achieve the purpose of lowering blood lipids. According to any embodiment of the plant fermentation liquid, it can greatly reduce the generation of calcium ion deposition in smooth muscle cells, thereby improving myocardial contractility and avoiding blood vessel blockage, thereby increasing metabolic rate to achieve lipid-lowering purposes. According to any embodiment of the plant fermentation liquid, it can increase the activity of vascular cell mitochondria, thereby improving the efficiency of blood vessels, and then increasing the metabolic rate to achieve the purpose of lowering lipids. According to any embodiment of the plant fermented liquid, it increases the body temperature, and then increases the metabolic rate to achieve the purpose of reducing fat. Moreover, according to any embodiment of the plant fermented liquid, the lipid-lowering purpose can be effectively achieved by taking 10 mL of the plant fermented liquid every day. Here, in some embodiments, the plant fermentation broth can be used to prepare a lipid-lowering composition.

In some embodiments, any of the aforementioned compositions can be a pharmaceutical. In other words, the medicinal product contains an effective amount of plant fermentation liquid.

In some embodiments, the foregoing pharmaceutical products may be formulated for parenteral, parenteral, oral, or topical administration using techniques well known to those skilled in the art. Dosage form.

In some embodiments, the dosage form for enteral or oral administration can be, but not limited to, tablet, troche, lozenge, pill, capsule , dispersible powder or granule, solution, suspension, emulsion, syrup, elixir, slurry or the like. In some embodiments, the dosage form for parenteral or topical administration may be, but not limited to, injection, sterile powder, external preparation, or the like. In some embodiments, the injection can be administered by subcutaneous injection, intraepidermal injection, intradermal injection or intralesional injection.

In some embodiments, the above-mentioned pharmaceuticals may include pharmaceutically acceptable carriers (pharmaceutically acceptable carriers) that are widely used in pharmaceutical manufacturing techniques. In some embodiments, the pharmaceutically acceptable carrier can be one or more of the following carriers: solvent, buffer, emulsifier, suspending agent, disintegrating agent ( decomposer), disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent ), gelling agents, preservatives, wetting agents, lubricants, absorption delaying agents, liposomes, and the like. The type and amount of carrier to be used is within the expertise and routine skill of those skilled in the art. In some embodiments, the solvent as the pharmaceutically acceptable carrier can be water, normal saline, phosphate buffered saline (PBS), or aqueous solution containing alcohol (aqueous solution containing alcohol).

In some embodiments, any of the foregoing compositions may be an edible product (ie, a food composition). In other words, the edible product contains a certain amount of vegetable broth. In some embodiments, the edible product can be general food, health food, dietary supplement or food additive.

In some embodiments, the aforementioned edible products can be manufactured into dosage forms suitable for oral administration using techniques well known to those skilled in the art. In some embodiments, general foods may be, but not limited to: beverages, fermented foods, bakery products or seasonings.

In some embodiments, the plant fermentation liquid of any embodiment can be added during raw material preparation (that is, as a food additive) by conventional methods, or the plant fermentation liquid of any embodiment can be added during the production of food (that is, as a food additive), and formulated with any edible material to be eaten by humans and non-human animals.

example 1 : Preparation of plant fermentation broth

First of all, prepare the raw materials of japonica seed, purslane raw material and water with a weight ratio of 2:1:80. Among them, the shelled fruit of the japonica procured from China is used as the raw material, and the leaf part of the purslane is used as the raw material of the purslane.

Next, mix the raw materials of kale seed, purslane raw material and water, and add 10% glucose relative to the total weight of the raw material of kale seed, purslane raw material and water to form a plant base solution. Herein, the Brix of the plant-based liquid is greater than 8.

The plant base liquid was heated to 95° C., and the heating was continued for 60 minutes after the heating environment reached 95° C. to obtain a water extract. Moreover, after the temperature of the water extract is lowered to less than 35° C., it can be used as a culture medium for subsequent fermentation procedures.

Add 0.1wt% beer yeast (Saccharomyces cerevisiae) and 0.05wt% lactic acid bacteria (Lactobacillus plantarum) to the culture medium, and culture it statically for three days to form primary fermentation liquid. Here, the brewer's yeast is the brewer's yeast with the registration number BCRC20271, and the lactic acid bacteria with the registration number BCRC 910805 is the TCI028 strain.

Then, add 5wt% acetic acid bacteria into the primary fermentation liquid and leave it to ferment for four days to form a plant fermentation stock solution. Herein, the acetic acid bacterium is the acetic acid bacterium with registration number BCRC11688. The brix degree of the plant fermentation stock solution is less than 4, and its acid-base value (pH value) is 4.5±0.5.

Filter the plant fermentation stock solution with a sieve with a pore size of 400 meshes, and carry out a vacuum concentration procedure on the filtered plant fermentation stock solution at 60°C and 150 bar, and add 60% isomaltooligosaccharides relative to the plant fermentation stock solution Afterwards, to obtain the plant fermentation liquid.

example 2 : Vascular cell mitochondrial activity test

Mitochondrion is a kind of organelle in the cell, which mainly synthesizes adenosine triphosphate (ATP) as a source of energy for the cell. When there are enough mitochondria in the cell, it usually means that the cell is younger and more elastic, and the metabolic rate of the cell is high. . Furthermore, since blood vessels are spread all over the body, venous endothelial cells are used as experimental cell lines to carry out experiments on blood vessel health and improvement of metabolic rate, so as to extend the effect of improving basal metabolism and lowering lipids.

2.1 Materials, instruments and solution configuration

Experimental cells: Human umbilical vein endothelial cells (HUVEC cells for short, registration number BCRC H-UV001).

Medium: EC medium (brand Gibco model M200) was used and supplemented with 10% low serum growth supplement (LSGS).

Buffer: phosphate buffered saline (Phosphate buffered saline, referred to as PBS, brand Gibco).

Detection instrument: flow cytometer (brand BD Accuri).

Detection reagent: MitoScreen JC-1 kit (MitoScreen JC-1 KIT, brand BD™), which includes JC-1 dye (BD™, MitoScreen) and JC-1 mitochondrial dye (mitochondrial dye).

2.2. Test process

Add 2 mL of the above medium to each well of a six-well plate and inoculate 1×10 ⁵ HUVEC cells, and divide the cells into three groups (namely blank group, control group and experimental group). Among them, the blank group does not add any test samples. The test sample added to the control group is the water extract formed during the preparation of Example 1. The test sample added to the experimental group is the plant fermentation stock solution prepared in Example 1. Here, the concentration of the test sample in the medium of the control group and the experimental group was 0.0625% (v/v).

After addition of side samples, groups were incubated at 37°C for 24 hours. After 24 hours, each group removed the medium, washed twice with 1x buffer, added trypsin and centrifuged at 400xg for 5 minutes, removed the supernatant, washed with 1mL of 1xPBS and centrifuged at 400xg for 5 minutes, then added 100µL The JC-1 dye was stained in the dark for 15 minutes, washed with 1mL of buffer and centrifuged at 400xg for 5 minutes, washed again with 1mL of washing buffer and centrifuged at 400xg for 5 minutes, added 500µL with 2 The 1x buffer solution of %FBS was used to suspend the cells, and then the membrane potential of the mitochondria of the cells was measured by flow cytometry to obtain the mitochondrial activity data. In order to avoid experimental operation errors, each group was operated twice with the same procedure, and the average value was taken. And, statistically significant differences between groups were statistically analyzed by Student's t-test, as shown in FIG. 1 . In Figure 1, "***" means that the p-value is less than 0.001 when compared with the blank group, and "**" means that the p-value is less than 0.01 when compared with the control group.

2.3. Test results

It can be seen from Figure 1 that after analysis and conversion, the relative mitochondrial activity of the blank group was 46.2%, the relative mitochondrial activity of the control group was 55.4%, and the relative mitochondrial activity of the experiment was 71.3%. Based on this, it can be seen that under the same culture conditions, after HUVEC cells were treated with water extracts of kale seed and purslane, compared with HUVEC cells without any treatment, the activity of mitochondria in the cells could be increased by 9.2 %.

Furthermore, after HUVEC cells were treated with the plant fermentation broth of kale seed and purslane, compared with HUVEC cells without any treatment, the activity of mitochondria in the cells could be increased by 25.1%.

Relatively speaking, after the HUVEC cells were treated with the plant fermentation liquid prepared from the kale seed and purslane, compared with the HUVEC cells treated with the water extract prepared from the kale seed and purslane, the intracellular The activity of the body can be increased by 15.9%. It can be seen that the fermentation process can effectively increase the content of active ingredients in the plant fermentation broth, so that the mitochondrial activity can be further improved.

example 3 : Vascular smooth muscle calcification test

In this test, β-glycerophosphate (β-glycerophosphate, hereinafter referred to as β-GP) was used to simulate the environment of hyperphosphatemia to induce calcification of cells, and to observe whether the plant fermentation broth could Effectively inhibit the occurrence of calcification.

3-1. Materials and solution configuration

Experimental cells: human aortic smooth muscle cells (Human aortic/smooth muscle cells, referred to as VSMC cells, using the cell line with the registration number ATCC Cat. CRL1999)

Buffer: 1x Dulbecco's Phosphate Buffered Saline, which is 1x DPBS purchased from Gibco model Cat. 14200-75 diluted 10 times with sterile double distilled water (ddH ₂ O).

1x Trypsin: Dilute 10x trypsin purchased from Gibco model Cat. 15400-054 10-fold with 1x DPBS buffer.

10 molar concentration (M) phosphatase inhibitor (β-glycerophosphate stock solution): 2.1604 g of β-glycerophosphate (β-glycerophosphate, β-GP, purchased from Sigma, model Cat. G9422) was dissolved in 1 mL of 1xDPBS buffer to a final concentration of 10M.

Cell culture medium: Dulbecco's Modified Eagle's Medium (DMEM, purchased from Gibco, 11965-092) was added with additional components to make it contain 10 vol% FBS (fetal bovine Serum, purchased from Gibco, 10437-028) and 1% Penicillin-Streptomycin (purchased from Gibco, Cat. 15140122).

Differentiation medium: 10 molar concentration of phosphatase inhibitor was diluted with cell culture medium to form differentiation medium containing 0 mM, 5 mM, 10 mM and 20 mM β-GP respectively.

10% formaldehyde solution: Dilute formaldehyde (formaldehyde, purchased from Jingming, model Cat. 119690010) 10 times with sterilized double distilled water.

Alizarin red dye: Dissolve 0.2g of Alizarin red (purchased from Sigma, model Cat.G9422) in 10mL of double distilled water, shake well and set aside.

3-2. Test process

Firstly, the initial cultivation of cells was carried out, and ^2x105 HASMC cells and 2000 µL cell culture medium were inoculated in each well of a six-well plate, and cultured in a carbon dioxide incubator for 24 hours, and the subsequent steps were performed after confirming the attachment of HASMC cells.

Next, according to the following Table 1, the cells were divided into eight groups and replaced with the experimental medium and then cultured and differentiated at 37°C for 14 days, wherein the experimental medium was also replaced every two days according to Table 1. Among them, the experimental medium of four groups is the differentiation medium without adding the test sample (i.e. the control group), and the experimental medium of the other four groups is the differentiation medium with the test sample added, and the added test sample is the plant prepared in Example 1 Fermentation stock solution (ie experimental group). Here, the concentration of the test sample added in the experimental group was 0.25% (v/v).

According to Table 1, the experimental group and the control group were further divided into the first control group, the second control group, the third control group, and the fourth control group according to the added concentration of β-GP, and the experimental group was also based on the concentration of β-GP. The added concentration is divided into the first experimental group, the second experimental group, the third experimental group, and the fourth experimental group.

Table 1 test group test sample β-GP concentration first control group none 0 mM The first experimental group Plant Fermentation Liquid 0 mM Second control group none 5mM The second experimental group Plant Fermentation Liquid 5mM The third control group none 10mM The third experimental group Plant Fermentation Liquid 10mM fourth control group none 20mM The fourth experimental group Plant Fermentation Liquid 20mM

After 14 days of culture, the liquid of each group was removed, and then washed once with buffer. Next, HASMC cells were fixed with 10% formaldehyde solution for 15 minutes. After removing the liquid of each group again, rinse once with double distilled water. Then, Alizarin Red stain was added for 5 minutes of staining. After the liquid of each group was removed again, it was rinsed twice with double distilled water, observed under a microscope and photographed and recorded as shown in FIG. 2 .

3-3. Test results

Please refer to FIG. 2 , in the first control group and the first experimental group, there is no visible calcified area, which means that the vascular cells are in a healthy and normal state. In the second control group, there was a local small-scale visible calcified area A, while in the second experimental group, there was no obvious visible calcified area, which means that the second control group had produced the precursor of initial vascular calcification, while the second experimental group The vascular cells still maintain a healthy and normal state.

In the third control group, there was a local large-scale visible calcification area A, while in the third experimental group there was a local small-scale visible calcification area A, which means that the third control group had vascular calcified lesions, while in the third experimental group Vascular cells were only a precursor to lesions where initial vascular calcification was seen and did not produce lesions.

In the fourth control group, there is a large-scale visible calcified area A, and the visible calcified area A occupies almost half of the picture, which means that the fourth control group has produced vascular calcified lesions, while the fourth experimental group still maintains local small The visible calcified area A means that severe vascular calcified lesions have occurred in the fourth control group, while vascular cells in the fourth experimental group have not yet produced lesions. It can be seen that the plant fermentation broth has a good inhibitory ability on arterial calcification, and can effectively maintain the healthy state of vascular cells even in the pathogenic environment simulated by high concentrations of phosphatase inhibitors. In other words, plant fermented liquid can effectively maintain the elasticity of blood vessels, maintain better blood circulation, and improve the body's metabolism.

example 4 : human test

4-1. Sample preparation

Plant fermentation broth: the plant fermentation broth prepared in Example 1 was used.

4-2. Subjects: 8 subjects (aged between 40 and 65 years old). Among them, each subject had at least one of the symptoms of systolic blood pressure > 120 mmHg, diastolic blood pressure > 80 mmHg, and total cholesterol > 200 mg/dL before the test. That is to say, this test selects middle-aged and high-aged subjects with poor metabolic rate and high probability of hyperlipidemia-related symptoms.

4-3. Test items: blood total cholesterol content, high-density lipoprotein content, low-density lipoprotein content, average fingertip temperature change, body fat and body mass index (BMI). Among them, the BMI is calculated by dividing the weight (kg) by the square of the height (meter).

4-4. Test method:

8 subjects were asked to drink 10mL of plant fermented liquid daily for 4 weeks. Before drinking (that is, the 0th week, also known as the control group) and after drinking for 4 weeks (that is, the 4th week, also known as the experimental group), the blood of the subjects was drawn respectively, and the blood test was carried out by Liren Medical Laboratory. For the detection of total cholesterol content, high-density lipoprotein content and low-density lipoprotein content, a portable high-precision infrared thermal imager (Testo 875-1i(N)) was used to measure the subjects before and after drinking for 15 Minutes and the average fingertip temperature of both hands after 4 weeks of continuous drinking, and the body fat meter (brand: TANITA, product: limbs and trunk body composition meter, model BC-545F) to measure the body weight of these subjects and Overall body fat percentage. Among them, the statistically significant difference between each group was statistically analyzed by Student's t-test. In Figure 3 to Figure 8, "*" means that the p-value is less than 0.05 compared with the control group, that is, there is a statistically significant difference between the experimental group and the control group.

4-5. Test results

Please refer to Figure 3. After four weeks of daily drinking of plant fermented liquid, the average cholesterol content of the 8 subjects decreased from 225.8 mg/dL (week 0) to 214.9 mg/dL (week 4), and the average total Cholesterol before and after the gap reached 4.8%. That is, drinking 10mL of plant fermented liquid daily can effectively reduce total cholesterol.

Please refer to Figure 4. After four weeks of daily drinking of plant fermented liquid, the average high-density lipoprotein levels in the blood of 8 subjects increased from 56.5mg/dL (week 0) to 60.2mg/dL (week 4 Weeks), the difference between before and after is 3.7mg/dL. That is, drinking 10mL of plant fermented liquid per day can significantly increase the high-density lipoprotein in the blood.

Please refer to Figure 5, after four weeks of daily drinking of plant fermented liquid, the average LDL content of 8 subjects decreased from 154.9mg/dL (week 0) to 140.9mg/dL (week 4), The difference between before and after is 14mg/dL. That is, drinking 10mL of plant fermented liquid every day can significantly reduce the concentration of low-density lipoprotein in the blood.

Please refer to Figure 6. After four weeks of daily drinking of plant fermented liquid, the average fingertip temperature of the 8 subjects increased from 31.7°C (week 0) to 32.7°C (week 4), with a difference of 1.0 ℃. That is to say, drinking 10mL of plant fermented liquid every day can significantly increase the average fingertip temperature.

Among them, there were three subjects whose average temperature at the tip of their fingers was only 23.6°C at week 0, that is to say, the peripheral blood circulation of these three subjects was worse than that of other subjects, and this After drinking 10mL of plant fermented liquid for 15 minutes, the three subjects felt that the temperature of the tip of the finger increased, and after measurement, the average temperature of the tip of the finger of the three test subjects rose to 25.9°C. In one of the three subjects, the temperature at the tip of the finger was 28.2°C before drinking the plant fermented liquid, and 15 minutes after drinking the plant fermented liquid, the temperature at the tip of the finger rose to 31.0°C, and the difference between before and after was even greater. up to 2.8°C. This means that after drinking 10mL of plant fermented liquid for 15 minutes, the temperature at the tips of the fingers can be significantly increased.

Please refer to Figure 7. After four weeks of daily drinking of plant fermented liquid, the average body fat of the 8 subjects decreased from 34.0% (week 0) to 31.3% (week 4), that is, the average body fat decreased by 2.7% . That is to say, drinking 10mL/day of plant fermented liquid can significantly reduce body fat.

Please refer to Figure 8, after four weeks of daily drinking of plant fermented liquid, the average body mass index of 8 subjects decreased from 28.1 kg/m ² (week 0) to 27.4 kg/m ² (week 4), That is, the average body mass index decreased by 0.7kg/m ² . That is, drinking 10mL/day of plant fermented liquid can significantly reduce the average body mass index.

It can be seen that long-term use of plant fermented liquid can improve body fat, lower blood fat, lower BMI index, increase finger tip temperature, etc., that is, plant fermented liquid has obvious effects of reducing fat and/or improving metabolism.

To sum up, according to any embodiment of the plant fermentation liquid, it can reduce cholesterol and low-density lipoprotein, and increase high-density lipoprotein, thereby reducing the blood fat content in the blood to achieve the purpose of lowering blood lipids. According to any embodiment of the plant fermentation liquid, it can greatly reduce the generation of calcium ion deposition in smooth muscle cells, thereby improving myocardial contractility and avoiding blood vessel blockage, thereby increasing metabolic rate to achieve lipid-lowering purposes. According to any embodiment of the plant fermentation liquid, it can increase the activity of vascular cell mitochondria, thereby improving the efficiency of blood vessels, and then increasing the metabolic rate to achieve the purpose of lowering lipids. According to any embodiment of the plant fermented liquid, it increases the body temperature, and then increases the metabolic rate to achieve the purpose of reducing fat. Moreover, according to any embodiment of the plant fermented liquid, the lipid-lowering purpose can be effectively achieved by taking 10 mL of the plant fermented liquid every day.

Although the technical content of the present invention has been disclosed above with preferred embodiments, it is not intended to limit the present invention. Any modification and modification made by those skilled in the art without departing from the spirit of the present invention should be covered by the present invention. Therefore, the scope of protection of the present invention should be defined by the scope of the appended patent application.

A: Visible areas of calcification

Figure 1 is a diagram of the experimental results of mitochondrial activity in vascular cells. Figure 2 is a graph showing the results of calcium ion deposition in smooth muscle cells. Fig. 3 is a graph showing the results of cholesterol content in blood in human experiments. Fig. 4 is a graph showing the results of high-density lipoprotein content in human experimental blood. Fig. 5 is a graph showing the results of low-density lipoprotein content in blood in human experiments. Figure 6 is a graph showing the results of average finger tip temperature after four weeks of human experimentation. Figure 7 is a graph showing the results of body fat changes in human experiments. Fig. 8 is a graph showing the change results of the average body mass index in the human experiment.

Claims (9)

A use of plant fermented juice, which is used to prepare a lipid-lowering composition, wherein the plant fermented juice is initially fermented by Saccharomyces cerevisiae and Lactobacillus plantarum, and It is obtained by secondary fermentation of acetic acid bacteria. The use as described in Claim 1, wherein the plant fermented juice is used for lowering cholesterol. The use as claimed in item 1, wherein the plant fermented juice is used to reduce low-density lipoprotein. The use as described in Claim 1, wherein the plant fermented juice is used to increase high-density lipoprotein. The use as claimed in item 1, wherein the plant fermented juice is used to reduce body fat. The use as claimed in item 1, wherein the plant fermented juice is used to reduce calcium ion deposition in smooth muscle cells. The use as described in Claim 1, wherein the plant fermented juice is used to enhance the activity of vascular cell granulocytes. The use as described in claim 1, wherein the plant fermented juice is used to increase the body temperature. The use as described in any one of claims 1 to 8, wherein the effective dosage of the plant fermented juice is 10mL/day.

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