CN111481594A

CN111481594A - Plant fermentation product and application thereof in regulating gene expression amount and cardiovascular health care

Info

Publication number: CN111481594A
Application number: CN201910357882.XA
Authority: CN
Inventors: 林咏翔; 庄伟秀
Original assignee: TCI Co Ltd
Current assignee: TCI Co Ltd
Priority date: 2019-01-25
Filing date: 2019-04-28
Publication date: 2020-08-04
Anticipated expiration: 2039-04-28
Also published as: CN111481594B; TW202028452A; TWI698521B

Abstract

The invention relates to the field of fermentation, in particular to a plant fermentation product and application thereof in regulating gene expression level and cardiovascular health care, and provides the plant fermentation product which is prepared by a method comprising the following steps of extracting a combination consisting of gardenia and purslane by using water to obtain a plant extract, and sequentially fermenting the plant extract with saccharomyces cerevisiae, lactobacillus plantarum and acetic acid bacteria to obtain the plant fermentation product.

Description

Plant fermentation product and application thereof in regulating gene expression amount and cardiovascular health care

Technical Field

The invention relates to the field of fermentation, in particular to a plant fermentation product and application thereof in regulating and controlling gene expression and cardiovascular health care.

Background

Cardiovascular diseases are a great threat to human health, and there is usually no obvious symptom at the beginning of onset, but by the time complications occur, such as cerebral stroke, myocardial infarction, heart failure, renal failure or retinal hemorrhage, the life of patients is usually seriously threatened, and the demand of cardiovascular medication for treating cardiovascular diseases is high, which becomes a huge burden of medical resources. Therefore, those skilled in the art are dedicated to develop products (including food products and pharmaceuticals) for protecting cardiovascular system, so as to meet the health needs of people.

However, most of the cardiovascular health products used at present are made of chemical components, which are not harmful to human health after long-term use, and these products are often expensive and not affordable for general users. In order to solve the above problems, those skilled in the art need to develop novel pharmaceutical products or food products with cardiovascular health efficacy to benefit the broad population in need thereof.

Disclosure of Invention

Accordingly, the present invention is directed to a plant fermented product prepared by a method comprising the steps of (a) extracting a combination of Gardenia (Gardenia jasminoides) and purslane (Portulaca oleracea) with water to obtain a plant extract, and (b) sequentially fermenting the plant extract with Saccharomyces cerevisiae, Lactobacillus plantarum (L Lactobacillus plantarum), and Acetobacter aceti to obtain the plant fermented product.

In an embodiment of the present invention, the fermentation time of the brewers' yeast and the germ lactic acid bacteria is 1 to 5 days, and the fermentation time of the acetic acid bacteria is 3 to 8 days.

In one embodiment of the invention, the effective concentration of the plant ferment is at least 2% (v/v).

In one embodiment of the present invention, the concentration of the Saccharomyces cerevisiae is 0.01-0.5% (v/v), the concentration of the Lactobacillus plantarum is 0.01-0.25% (v/v), and the concentration of the acetic acid bacteria is 1-20% (v/v).

In an embodiment of the present invention, the volume ratio of the gardenia to the purslane to the water is 1-5: 1: 40-60.

Another objective of the present invention is to provide a plant fermentation product for preparing a composition for regulating CD36 gene, ATP binding cassette family A member 1(ATP binding cassette family A member1, ABCA1) gene, protein C (PROC) gene, Weatherburg blood coagulation factor (von Willebrand factor, VWF) gene, F3 gene, serine protease inhibitor family E member1 (serum protease inhibitor E member1, PIPINE 1) gene, platelet derived growth factor C (platelet derived growth factor C, PDGFC) gene, fibroblast growth factor 2(fibroblast growth factor 2, FGF2) gene, Insulin-like growth factor 2mRNA binding protein 3(Insulin-like protein 2-binding protein 3, Saccharopolyspora sinensis receptor 3) gene, and a cell growth factor receptor SP (Portulaceae) expressing protein C1, Saccharomyces cerevisiae 3, Saccharomyces cerevisiae 1, Saccharomyces cerevisiae 3, Saccharomyces cerevisiae 1, Saccharomyces cerevisiae 1, Saccharomyces cerevisiae 3, Saccharomyces cerevisiae, Lactobacillus, and Lactobacillus acidi, Saccharomyces cerevisiae, and Lactobacillus acidi, wherein the plant growth factor 3, Saccharomyces cerevisiae, Lactobacillus acidi are sequentially fermented by fermentation to obtain a cell, and a cell growth factor, and a cell growth factor for preparing a cell, and a cell growth factor for fermentation composition for preparing a cell.

Another object of the present invention is to provide a use of a plant fermentation product for preparing a cardiovascular health composition, wherein the plant fermentation product is prepared by a process comprising the steps of (a) extracting a combination of Gardenia jasminoides and Portulaca oleracea with water to obtain a plant extract, and (b) sequentially fermenting the plant extract with Saccharomyces cerevisiae, Lactobacillus plantarum L, and Acetobacter aceti to obtain the plant fermentation product.

In an embodiment of the present invention, the volume ratio of the gardenia, the purslane and the water is 1-5: 1: 40-60.

In one embodiment of the present invention, the composition is in the form of a pharmaceutical or a food product.

In conclusion, the plant fermentation product has the effects of regulating the expression levels of a CD36 gene, an ABCA1 gene, a PROC gene, a VWF gene, an F3 gene, a SERPINE1 gene, a PDGFC gene, an FGF2 gene, an IGF2BP3 gene, an IGF1R gene, an I L8 gene, a VCAM1 gene and a CASP8 gene, regulating the whole cardiac meridian in five-element meridians, regulating the expression of vasodilatation and atherosclerosis genes, reducing thrombosis, regulating the angiogenesis genes and blood pressure, reducing the vascular inflammation genes and the hemagglutination reaction and achieving the cardiovascular health care effect.

The following examples are presented to illustrate the present invention and are not intended to limit the scope of the invention, which is not to be construed as limiting the scope of the invention since various modifications and changes may be made by those skilled in the art without departing from the spirit and scope of the invention.

Drawings

FIG. 1 is a data graph showing the measurement of total flavone content in the fermented plant material of the present invention;

FIG. 2 is a graph of data on the efficacy of the plant fermentations of the invention in modulating the expression of genes associated with atherosclerosis (including the CD36 gene and the ABCA1 gene), wherein ". x" indicates that p < 0.05 compared to a control; ". indicates p < 0.01, compared to control.

Fig. 3 is a graph of data on the efficacy of the plant fermentations of the invention in modulating expression of cardiovascular health related genes (including the PROC, VWF, F3, and SERPINE1 genes), wherein "×" indicates p < 0.001 compared to a control group;

FIG. 4 is a graph of data showing the efficacy of plant fermentations of the present invention in modulating expression of cardiovascular health related genes including the PDGFC, FGF2, IGF2BP3, and IGF1R genes, wherein ". x" indicates p < 0.05 compared to a control; "x" indicates p < 0.001 compared to control;

FIG. 5 is a graph of data on the efficacy of the plant fermentations of the present invention in modulating expression of genes associated with cardiovascular health (including the I L8 and VCAM1 genes), wherein ". star" indicates p < 0.05 as compared to a control group, ". star" indicates p < 0.01 as compared to a control group, ". star" indicates p < 0.001 as compared to a control group;

fig. 6 is a data plot of the efficacy of the plant fermentations of the present invention in modulating expression of cardiovascular health related genes (i.e., CASP8 gene), wherein "×" indicates p < 0.001 as compared to the control group.

Detailed Description

Definition of

As used herein, the numerical values are approximations and all numerical data are reported to be within the 20 percent range, preferably within the 10 percent range, and most preferably within the 5 percent range.

Statistical analysis was performed using Excel software. Data are presented as mean ± Standard Deviation (SD), and differences between individual data are analyzed by student's t-test (student's t-test).

According to the present invention, Gardenia (Gardenia jasminoides) is an evergreen shrub of the genus Gardenia (Gardenia) of the family Rubiaceae (Rubiaceae), also known as Plumbum preparatium and Denmark Calophyllum. Fructus gardeniae is used as a medicine in the traditional Chinese medicine, is cold in nature and bitter in taste, has the functions of clearing heat and purging fire, and is mainly used for treating conjunctival congestion, jaundice, hematemesis and heat-toxin pyocutaneous disease.

According to the invention, purslane (Portulaca oleracea) is a herb of Portulacaceae (Portulacaceae) Portulaca genus (Portulaca), which is known as purslane, Portulaca oleracea and five-element grass. The purslane is used as a medicine in the traditional Chinese medicine by using overground whole herbs, is sweet and sour in taste, cold in nature and non-toxic, and has the effects of clearing heat and removing toxicity, eliminating dampness and stopping dysentery, promoting urination and moistening lung, quenching thirst and promoting fluid production.

The procedures and parameters involved in the fermentation and culture according to the present invention are within the skill of those skilled in the art.

As used herein, the terms "Saccharomyces cerevisiae", "Lactobacillus plantarum" (L Acetobacter plantarum) "and" Acetobacter aceti "are intended to encompass Saccharomyces cerevisiae, Lactobacillus plantarum, and Acetobacter aceti, respectively, which are readily available to one of ordinary skill in the art (e.g., from domestic or foreign depositories), or strains of Saccharomyces cerevisiae, Lactobacillus plantarum, and Acetobacter aceti, respectively, that have been isolated and purified from natural sources using microbial isolation methods customary in the art.

In accordance with the present invention, the pharmaceutical may be manufactured in a dosage form (dosage form) suitable for parenteral (parenteral) or oral (oral) administration using techniques well known to those skilled in the art, including, but not limited to: injection (injection) [ for example, sterile aqueous solution (sterile aqueous solution) or dispersion (dispersion) ], sterile powder (sterile powder), troche (tablet), tablet (troche), buccal tablet (dosage), pill (pill), capsule (capsule), dispersible powder (dispersible powder) or granule (granule), solution, suspension (suspension), emulsion (emulsion), syrup (syrup), elixir (elixir), syrup (slurry), and the like.

The medicament according to the invention may be administered in parenteral routes (parenteral routes) selected from the group consisting of: intraperitoneal injection (intraperitoneal injection), subcutaneous injection (subcutaneous injection), intramuscular injection (intramuscular injection), and intravenous injection (intravenous injection).

The pharmaceutical according to the present invention may comprise a pharmaceutically acceptable carrier (pharmaceutically acceptable carrier) which is widely used in pharmaceutical manufacturing technology. For example, the pharmaceutically acceptable carrier may comprise one or more agents selected from the group consisting of: solvents (solvents), emulsifying agents (emulsifiers), suspending agents (suspending agents), disintegrating agents (disintegrants), binding agents (binders), excipients (excipients), stabilizing agents (stabilizing agents), chelating agents (chelating agents), diluents (diluents), gelling agents (gelling agents), preservatives (preserving), lubricants (lubricants), absorption delaying agents (absorption delaying agents), liposomes (lipids), and the like. The selection and amounts of such agents are within the skill and routine skill of those skilled in the art.

According to the present invention, the pharmaceutically acceptable carrier comprises a solvent selected from the group consisting of: water, normal saline (normal saline), Phosphate Buffered Saline (PBS), sugar-containing solutions, aqueous alcohol-containing solutions (aqueous solution stabilizing alcohol), and combinations thereof.

According to the present invention, the food product can be used as a food additive (food additive) to be added during the preparation of the raw material or during the preparation of the food by conventional methods, and can be formulated with any edible material into a food product for human and non-human animal consumption.

According to the present invention, the types of food products include, but are not limited to: beverages (leafages), fermented foods (fermented foods), bakery products (bakery products), health foods (health foods) and dietary supplements (dietary supplements).

Example 1 preparation of plant fermentations

Firstly, Gardenia (Gardenia jasminoides), purslane (Portulaca oleracea) and water from China are mixed in a volume ratio of 1-5: 1: 40-60, and are sterilized and extracted for 0.5-3 hours at 50-100 ℃ respectively to obtain a plant extract, the plant extract is cooled to room temperature for subsequent three-stage fermentation, the three-stage fermentation is to sequentially inoculate 0.01-0.5% (v/v) brewer's yeast (Saccharomyces cerevisiae) BCRC 20271 and 0.01-o.25% (v/v) germ lactic acid bacteria (L active bacteria) TCI028(BCRC 910805) for fermentation for 1-5 days, 1-20% (v/v) acetic acid bacteria (Acetobacter aceti) BCRC 88 (the above strains are all obtained from Research Institute of Food Industry, Research Institute of Taiwan (Research Institute), and Research Institute of Food Industry, Research, fermentation for 1-5 days, and filtration of fermentation of malt oligosaccharide (Research, and fermentation for 400-40-200-mesh fermentation, and filtration of the above strains.

Example 2 detection of Total Flavonoids content of plant fermentations

The experimental procedure for detecting the content of total flavonoids is as follows, the content of total flavonoids is expressed by taking rutin (chromadexsb-00018440) equivalent as the relative content of total flavonoids, and the preparation material comprises 10% aluminum nitrate (aqueous solution) (Alfa Aesar 12360), 5% sodium citrate (aqueous solution) (Sigma31443), 4% sodium hydroxide (aqueous solution) (Macron 7708-10) and 200 μ g/m L of rutin (methanol solution).

Adding the

standard liquid

0, 200 μ L, 400 μ L, 600 μ L0, 800 μ L1, 1000 μ L2 and 1200 μ L3 into a test tube, adding water 1200 μ L4, 1000 μ L5, 800 μ L6, 600 μ L7, 400 μ L, 200 μ L and 0 μ L, shaking, mixing, adding 200 μ L-containing rutin solution, adding 200 μ L sodium citrate 5%, mixing, standing for 6 min, adding 200 μ L-10% aluminum nitrate, mixing, standing for 6 min, adding 2m L-containing sodium hydroxide 4%, mixing, and adding 1.4m L H₂Mixing O, placing 200 μ L above reaction solution in 96-well reaction plate, detecting absorbance at 500nm with spectrophotometer, and drawing standard curve.

The plant fermentation product obtained in example 1 was used as an experimental group, and the plant extract was used as a comparative group, and the experimental group or the comparative group was diluted appropriately, and the 200 μ L experimental group or the comparative group was placed in a test tube, and 200 μ L5% sodium citrate was added, mixed well and left to stand for 6 minutes, 200 μ L10% aluminum nitrate was added, mixed well and left to stand for 6 minutes, 2m L4% sodium hydroxide was added, mixed well and then 1.4m L H was added₂Mixing O, placing 200 μ L above reaction solution in 96-well reaction plate, detecting absorbance at 500nm with spectrophotometer, and showing the result of total flavone content in FIG. 1.

FIG. 1 is a data graph showing the measurement of the total flavone content in the fermented plant material of the present invention. As can be seen from FIG. 1, the total flavone content in the experimental group was significantly increased (1.4 times) compared to the comparative group. The results of this example show that the plant fermentations of the invention release large amounts of total flavonoids.

Example 3 evaluation of the effectiveness of plant fermentations in cardiovascular health

3.1 efficacy of plant fermentations in regulating Gene expression associated with atherosclerosis

First, human monocytic cells (human monoclonal cells) THP-1 (corresponding to ATCC, TIB202) were cultured in RPMI-1640 medium (Gibco) supplemented with 10% Fetal Bovine Serum (FBS) (Gibco), 0.05mM 2-mercaptoethanol (2-mercaptethanol), 100 units/m L penicillin (penillimin) and 100. mu.g/L streptomycin (streptomycin). 2m L of medium was added to each well of a 6-well plate to have 1.5X 10 per well⁵A THP-1 cell. Next, a differentiation medium with 500nM phorbol-12-tetradecanoyl-13-acetate (phorbol 12-myrisitate 13-acetate, PMA) was added and cultured for 48 hours. Thereafter, the medium was replaced with fresh one and further cultured for 48 hours.

In addition, one-way plant fermentates (including single purslane fermentate, single gardenia fermentate) were prepared as comparative analysis samples, and the preparation was performed according to the procedure of example 1 above, except that: the composition comprising fructus Gardeniae and herba Portulacae is replaced by herba Portulacae and fructus Gardeniae.

Then, THP-1 cells were divided into 5 groups including 1 experimental group, 2 comparative groups (i.e., comparative group 1 and comparative group 2), 1L PS (lipopolysaccharide, L ipolysochichorides) group and 1 control group.2% (v/v) of the plant fermentation according to example 1 above was added to the cells of the experimental group, 2% (v/v) of the purslane fermentation was added to the cells of

comparative group

1, 2% (v/v) of the gardenia fermentation was added to the cells of comparative group 2, and 100ng/m L lipopolysaccharide (L ipolysochichorides, L PS) was added to the cells of L PS group to induce a cell inflammation reaction, which was related to vascular dysfunction (vasculr dysfuntion). The cells of the control group were not treated.

In this example, genes for analysis of atherosclerosis include the CD36 gene and the ATP binding cassette family A member 1(ATP binding cassette subset A member1, ABCA1) gene.

The respective cell cultures obtained above were subjected to RNA extraction using an RNA extraction kit (Genemark). 2,000ng of each RNA group thus obtained was extracted and added

III reverse transcriptase (Invitrogen) reverse transcribes the extracted RNA to cDNA. Then, using cDNA as a template and primer pairs for amplifying a target gene, including CD36, ABCA1 and ACTB (as internal control groups), the nucleotide sequences of which are shown in table 1 below, quantitative real-time PCR was performed using KAPA CYBR FAST qPCR kit (2x) (KAPA Biosystems) in StepOne Plus real-time PCR system (ABI) to amplify and quantify the target gene. The melting curve of the PCR product was confirmed during the quantitative real-time PCR reaction.

TABLE 1

The relative expression of the target gene is derived from the equation

And calculating relative fold change using cycle thresholds and standard deviations for the ACTB gene (as an internal control group) and the reference gene, where Δ Ct ═ Ct_{Target/reference genes}-Ct_ACTB，ΔΔCt＝ΔCt_{Target gene}-ΔCt_{Reference gene}，

The expression level of the target gene in the control group was used as a comparative reference for 1. Statistically significant differences between groups were determined by the single-tailed Steady's t-assay. The results of this example are shown in FIG. 2.

FIG. 2 is a data diagram showing the efficacy of the plant fermentation product of the present invention in regulating the expression of the genes related to atherosclerosis (including CD36 gene and ABCA1 gene). As can be seen from FIG. 2, the fold change of the genes in the experimental group is significantly reduced by about 70% in the case of the CD36 gene compared with the control group, L PS group, comparative group 1 and comparative group 2, and is significantly increased by about 100% in the case of the ABCA1 gene compared with the control group, L PS group, comparative group 1 and comparative group 2.

3.2 efficacy of plant fermentations in regulating Gene expression associated with cardiovascular health

First, human monocytic cells (human monoclonal cells) THP-1 (corresponding to ATCC, TIB202) were cultured in RPMI-1640 medium (Gibco) supplemented with 10% Fetal Bovine Serum (FBS) (Gibco), 0.05mM 2-mercaptoethanol (2-mercaptethanol), 100 units/m L penicillin (penillimin) and 100. mu.g/L streptomycin (streptomycin). 2m L of medium was added to each well of a 6-well plate to have 1.5X 10 per well⁵A THP-1 cell. Then, add the utensilDifferentiation medium with 500nM phorbol-12-tetradecanoyl-13-acetate (phorbol 12-myrisitate 13-acetate, PMA) and 48 hours of culture. Thereafter, the medium was replaced with fresh one and further cultured for 48 hours.

Then, the THP-1 cells were divided into 4 groups, including 1 experimental group, 2 comparative groups (i.e., comparative group 1 and comparative group 2), and 1 control group. 0.5% (v/v) of the plant fermentation according to example 1 above was added to the cells of the experimental group, 0.5% (v/v) of the purslane fermentation was added to the cells of the comparative group 1, and 0.5% (v/v) of the gardenia fermentation was added to the cells of the comparative group 2. Cells of the control group were not treated. Subsequently, each set of cells was cultured in an incubator for 24 hours, and then the cells were treated with a cell lysis buffer (cell lysis buffer), and then each set of cell cultures was harvested and taken for gene expression analysis.

In this example, genes related to cardiovascular health include protein C (PROC) gene, wen Willebrand factor (VWF) gene, F3 gene, serine protease inhibitor family E member 1(serine protease inhibitor family E member1, SERPINE1) gene, platelet-derived growth factor C (platelet derived growth factor C, PDGFC) gene, fibroblast growth factor 2(fibroblast growth factor 2, FGF2) gene, Insulin-like growth factor 2mRNA binding protein 3(Insulin-like growth factor 2 mRNA-binding protein 3, IGF2BP3) gene, Insulin-like growth factor 1receptor (Insulin growth factor 1 ke, receptor R) gene, Interleukin 1 mediator 8 (Interleukin-leukocyte protease) gene, vascular adhesion promoter 3 (vccell kinase) gene, IGF2BP 598, and cell adhesion promoter 398 (cell adhesion gene, vccell adhesion promoter 3, vccell kinase 8, vccell adhesion promoter 3, and vccell adhesion promoter 398 gene.

The top was set with RNA extraction kit (Genemark)The resulting cell cultures of each group were subjected to RNA extraction. 2,000ng of each RNA group thus obtained was extracted and added

III reverse transcriptase (Invitrogen) reverse transcribes the extracted RNA into cDNA, then cDNA is used as a template and primer pairs for amplifying the target gene, including PROC, VWF, F3, SERPINE1, PDGFC, FGF2, IGF2BP3, IGF1R, I L8, VCAM1, CASP8, and ACTB (as internal control groups), whose nucleotide sequences are shown in table 2 below, are used to perform quantitative real-time PCR in the StepOne Plus real-time PCR system (ABI) using the KAPA CYBR FAST qPCR kit (2x) (KAPA Biosystems) to amplify and quantify the target gene.

TABLE 2

The relative expression of the target gene is derived from the equation

The expression level of the target gene in the control group was used as a comparative reference for 1. Statistically significant differences between groups were determined by the single-tailed Steady's t-assay. The results of this example are shown in fig. 3-6.

FIGS. 3 to 6 are graphs showing the efficacy of the fermented plant material of the present invention in controlling the expression of cardiovascular health-related genes (including PROC, VWF, F3, SERPINE1, PDGFC, FGF2, IGF2BP3, IGF1R, I L, VCAM1, and CASP8 genes). As can be seen from FIG. 3, the relative expression of the genes in the experimental group was significantly increased as compared with the control group, comparative group 1, and comparative group 2 by about 320% as compared with the control group, and significantly decreased as compared with the control group, comparative group 1, and comparative group 2 by about 70% as compared with the PROC gene, and significantly decreased as compared with the control group, comparative group 1, and comparative group 2 by about 75% as compared with the SERPINE1, and thus significantly decreased by about 20% as compared with the control group, and 1% as compared with the SERPINE 1.

As can be seen from FIG. 4, the PDGFC gene expression was significantly reduced compared to the control, comparative 1 and comparative 2, wherein the relative expression of the gene was reduced by about 75% compared to the control; for the FGF2 gene, the relative expression of the genes in the experimental group was significantly reduced compared to the control group, comparative group 1 and comparative group 2, wherein the relative expression was reduced by about 30% compared to the control group; for IGF2BP3 gene, the relative expression of the genes in the experimental group was significantly reduced compared to the control group, comparative group 1 and comparative group 2, wherein the relative expression was reduced by about 60% compared to the control group; in the case of IGF1R gene, the relative expression of the genes in the experimental group was significantly reduced compared to the control group, comparative group 1 and comparative group 2, wherein the relative expression was reduced by about 45% compared to the control group.

As can be seen from FIG. 5, the relative expression level of the genes in the experimental group was significantly reduced compared to the control group, comparative group 1 and comparative group 2 by about 45% for the gene I L8, and by about 60% for the gene VCAM 1.

As can be seen from FIG. 6, the relative expression of genes in the experimental group was significantly reduced with respect to the CASP8 gene as compared with the control group, comparative group 1 and comparative group 2, wherein the relative expression was reduced by about 60% as compared with the control group.

The experimental result shows that the plant fermentation product can inhibit thrombosis by improving PROC gene expression, reduce thrombosis by inhibiting VWF, F3 and SERPINE1 gene expression, inhibit vasoconstriction caused by vascular cell proliferation and regulate blood pressure by inhibiting PDGFC, FGF2, IGF2BP3 and IGF1R gene expression, and reduce blood coagulation generation by inhibiting I L8, VCAM1 and CASP8 gene expression.

In conclusion, the plant fermentation product can regulate the expression levels of a CD36 gene, an ABCA1 gene, a PROC gene, a VWF gene, an F3 gene, a SERPINE1 gene, a PDGFC gene, an FGF2 gene, an IGF2BP3 gene, an IGF1R gene, an I L8 gene, a VCAM1 gene and a CASP8 gene, regulate the whole cardiac meridian in five rows of meridians, regulate the expression of vasodilation and atherosclerosis genes, reduce thrombosis, regulate the expression of angiogenesis genes and blood pressure, reduce vascular inflammation genes and blood coagulation reaction, and achieve the effect of cardiovascular health care.

The foregoing is by way of example only, and not limiting. It is intended that all equivalent modifications or variations without departing from the spirit and scope of the present invention shall be included in the appended claims.

Claims

1. A plant fermentation product prepared by a process comprising the steps of:

(a) extracting a combination of Gardenia jasminoides and Portulaca oleracea with water to obtain a plant extract; and

(b) the plant extract is sequentially fermented with Saccharomyces cerevisiae, germ lactobacillus (L Acetobacter plantarum) and acetic acid bacteria (Acetobacter aceti) to obtain the plant fermentation product.

2. The fermented plant material of claim 1, wherein the fermentation time of the brewer's yeast and the germ lactic acid bacteria is 1 to 5 days, and the fermentation time of the acetic acid bacteria is 3 to 8 days.

3. The plant fermentate according to claim 1, wherein the effective concentration of the plant fermentate is at least 2% (v/v).

4. The fermented plant material according to claim 1, wherein the concentration of the brewer's yeast is 0.01-0.5% (v/v), the concentration of the germ lactic acid bacteria is 0.01-0.25% (v/v), and the concentration of the acetic acid bacteria is 1-20% (v/v).

5. The plant fermentation product of claim 1, wherein the volume ratio of the gardenia, the purslane and the water is 1-5: 1: 40-60.

6. A plant fermentation product for preparing a composition for regulating CD36 gene, ATP binding cassette family a member1 (ABCA 1) gene, protein C (PROC) gene, wenwebber's blood coagulation factor (VWF) gene, F3 gene, serine protease inhibitor family E member 1(serine protease inhibitor family E member1, SERPINE1) gene, platelet-derived growth factor C (platelet-derived growth factor C, PDGFC) gene, fibroblast growth factor 2(fibroblast growth factor 2) gene, Insulin-like growth factor 2mRNA binding protein 3(Insulin-like growth factor 2-mRNA-binding protein 3, IGF 2-like 3 gene, Insulin-like growth factor 1 (Insulin receptor) gene, wherein the expression of protein C (vegf-binding cassette) gene, vegf-binding protein 3, IGF1 gene, IGF-binding protein 3, IGF-protease receptor (IGF 8) gene, and the expression of protein C (VWF) gene, vegf-binding cassette, VWF-binding protein C, VWF-C, vegf-protein C, vegf-1, vegf-binding protein C, vegf-1, vegf-binding protein 3, IGF-1, IGF-5, wherein the expression of the gene is expressed by fermentation product of CD-binding protein C, wherein the expression of CD-binding protein C, the expression of CD-binding protein C, the gene, the expression of CD-protein C, the gene, the expression of:

7. Use of a plant fermentation for the preparation of a cardiovascular health composition, wherein said plant fermentation is prepared by a process comprising the steps of:

(a) extracting a combination of Gardenia jasminoides Ellis (Gardenia jasminoides) and Portulaca oleracea (Portulaca oleracea) with water to obtain a plant extract; and

8. The use according to claim 6 or 7, wherein the fermentation time of the brewer's yeast and the germ lactic acid bacteria is 1 to 5 days, and the fermentation time of the acetic acid bacteria is 3 to 8 days.

9. Use according to claim 6 or 7, wherein the effective concentration of the plant ferment is at least 2% (v/v).

10. The use according to claim 6 or 7, wherein the concentration of Saccharomyces cerevisiae is 0.01-0.5% (v/v), the concentration of Lactobacillus plantarum is 0.01-0.25% (v/v), and the concentration of Acetobacter xylinum is 1-20% (v/v).

11. The use of claim 6 or 7, wherein the volume ratio of the gardenia, the purslane and the water is 1-5: 1: 40-60.

12. Use according to claim 6 or 7, wherein said composition is in the form of a medicament or a food product.