TWI698521B - Plant fermentation product and uses of the plant fermentation product for regulating expressions of cd36, abca1, proc, vwf, f3, serpine1, pdgfc, fgf2, igf2bp3, igf1r, il8, vcam1, and casp8 genes, and cardiovascular care - Google Patents

Abstract

The present disclosure provides a plant fermentation product and uses of the plant fermentation product for regulating expressions of CD36, ABCA1, PROC, VWF, F3, SERPINE1, PDGFC, FGF2, IGF2BP3, IGF1R, IL8, VCAM1, and CASP8 genes, and cardiovascular care.

Description

Plant fermented products and their use for regulating the expression levels of CD36 gene, ABCA1 gene, PROC gene, VWF gene, F3 gene, SERPINE1 gene, PDGFC gene, FGF2 gene, IGF2BP3 gene, IGF1R gene, IL8 gene, VCAM1 gene and CASP8 gene and Uses for cardiovascular health

The present invention relates to a plant fermented product and its use for regulating CD36 gene, ABCA1 gene, PROC gene, VWF gene, F3 gene, SERPINE1 gene, PDGFC gene, FGF2 gene, IGF2BP3 gene, IGF1R gene, IL8 gene, VCAM1 gene and The expression level of CASP8 gene and the use of cardiovascular health care.

Cardiovascular disease is a major threat to human health. According to the data released by the Ministry of Health and Welfare of the Republic of China, cardiovascular disease accounts for three of the top ten causes of death for Chinese people. Among them, heart disease and cerebrovascular disease are ranked second and second respectively. 4. In the United States, cardiovascular disease is the number one cause of death for the American people. It can be seen that the threat of cardiovascular disease to human health has reached a level that cannot be ignored. There are usually no obvious symptoms at the beginning of its onset, but it will wait until once it appears. In the event of complications, such as stroke, myocardial infarction, heart failure, renal failure or retinal hemorrhage, the patient’s life is usually severely threatened. In order to treat cardiovascular diseases, the demand for cardiovascular drugs is also high, becoming a medical resource Huge burden. Therefore, those skilled in the art are committed to developing products (including food products and pharmaceuticals) for protecting the cardiovascular system to meet the needs of the general human health.

However, most of the currently used cardiovascular health care products are made of chemical ingredients. Long-term use is not only harmful to human health, but also these products are often expensive and not affordable for ordinary users. In order to solve the above-mentioned problems, those skilled in the art urgently need to develop novel medicines or food products with cardiovascular health effects to benefit the vast populations in need.

In view of this, the object of the present invention is to provide a plant fermented product, which is prepared by a method including the following steps: (a) Extracting a sesame seed ( Gardenia jasminoides ) and a purslane ( Portulaca oleracea ) with water And (b) the plant extract is fermented with Saccharomyces cerevisiae , Lactobacillus plantarum and Acetobacter aceti in sequence to obtain The fermented plant.

In an embodiment of the present invention, 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.

In an embodiment of the present invention, the effective concentration of the plant fermented product is at least 2% (v/v).

In one embodiment of the present invention, the concentration of the brewer's yeast is between 0.01 and 0.5% (v/v), the concentration of the germ lactic acid bacteria is between 0.01 and 0.25% (v/v), and the concentration of the acetic acid bacteria Between 1~20%(v/v).

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

Another object of the present invention is to provide a plant fermented product for preparing a regulatory CD36 gene, ATP binding cassette subfamily A member 1 (ATP binding cassette subfamily A member 1, ABCA1 ) gene, protein C ( PROC ) gene, Von Willebrand factor ( VWF ) gene, F3 gene, serine protease inhibitor family E member 1 (serine protease inhibitor family E member 1, SERPINE1 ) gene, platelet derived growth factor C (platelet derived growth factor) C, PDGFC ) gene, fibroblast growth factor 2, FGF2 gene, Insulin-like growth factor 2 mRNA-binding protein 3 ( IGF2BP3 ) gene, insulin-like Growth factor 1 receptor (insulin like growth factor 1 receptor, IGF1R ) gene, interleukin-8 ( IL8 ) gene, vascular cell adhesion molecule 1 ( VCAM1 ) gene and apoptosis protease 8 (caspase 8, CASP8 ) gene expression composition, wherein the plant fermented product is prepared by a method including the following steps: (a) Extraction of gardenia jasminoides and horses with water A combination of Portulaca oleracea to obtain a plant extract; and (b) the plant extract is combined with Saccharomyces cerevisiae , Lactobacillus plantarum and Acetobacter aceti The fermentation is carried out sequentially to obtain the plant fermented product.

Another object of the present invention is to provide a use of a plant fermented product for the preparation of a cardiovascular health care composition, wherein the plant fermented product is prepared by a method including the following steps: (a) Extraction with water-from A combination of Gardenia jasminoides and Portulaca oleracea to obtain a plant extract; and (b) the plant extract and Saccharomyces cerevisiae and Lactobacillus plantarum And Acetobacter aceti are fermented in sequence to obtain the plant fermented product.

In an embodiment of the present invention, 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.

In an embodiment of the present invention, the effective concentration of the plant fermented product is at least 2% (v/v).

In one embodiment of the present invention, the concentration of the brewer's yeast is between 0.01 and 0.5% (v/v), the concentration of the germ lactic acid bacteria is between 0.01 and 0.25% (v/v), and the concentration of the acetic acid bacteria Between 1~20%(v/v).

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

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

In summary, the effect of the plant fermented product of the present invention is that it can regulate CD36 gene, ABCA1 gene, PROC gene, VWF gene, F3 gene, SERPINE1 gene, PDGFC gene, FGF2 gene, IGF2BP3 gene, IGF1R gene, and IL8 gene. , VCAM1 gene and CASP8 gene expression level, regulate the entire heart meridian in the five elements meridian, regulate vasodilation and atherosclerosis gene expression, reduce thrombosis, regulate angiogenesis genes, relax blood pressure, and reduce vascular inflammation genes and reduce coagulation response , To achieve the effect of cardiovascular health care.

The following will further explain the embodiments of the present invention. The following examples are used to illustrate the present invention and are not intended to limit the scope of the present invention. Anyone familiar with the art will not depart from the spirit and scope of the present invention. Some changes and modifications can be made, so the scope of protection of the present invention shall be subject to the scope of the attached patent application.

Figure 1 is a data diagram of the total flavonoid content of the fermented plant of the present invention.

Fig. 2 is a data chart showing the efficacy of the plant fermented product of the present invention in regulating the expression of genes related to atherosclerosis (including CD36 gene and ABCA1 gene), where "*" indicates comparison with the control group, p <0.05;"**" indicates that p <0.01 compared with the control group.

Figure 3 is a data diagram showing the efficacy of the plant fermented product of the present invention in regulating the performance of genes related to cardiovascular health care (including PROC, VWF, F3 , and SERPINE1 genes), where "***" means comparison with the control group, p <0.001.

Figure 4 is a data diagram showing the efficacy of the plant fermented product of the present invention in regulating the performance of genes related to cardiovascular health care (including PDGFC, FGF2, IGF2BP3 , and IGF1R genes), where "*" indicates comparison with the control group, p <0.05 ; "***" means compared with the control group, p <0.001.

Fig. 5 is a data diagram of the efficacy of the plant fermented product of the present invention in regulating the performance of genes related to cardiovascular health care (including IL8 and VCAM1 genes), where "*" indicates comparison with the control group, p <0.05;"**" Means compared with the control group, p <0.01;"***" means compared with the control group, p <0.001.

Fig. 6 is a data diagram showing the efficacy of the plant fermented product of the present invention in regulating the performance of genes related to cardiovascular health care (ie, CASP8 gene), where "***" means compared with the control group, p <0.001.

definition

The numerical values used herein are approximate values, and all experimental data are expressed in the range of 20%, preferably in the range of 10%, and most preferably in the range of 5%.

Use Excel software for statistical analysis. Data mean ± standard deviation (SD) represented by the difference between the two herein by Student t test (student's t -test) analysis.

According to the present invention, Gardenia jasminoides is an evergreen shrub belonging to the Rubiaceae (Rubiaceae) genus Gardenia , also known as Molitan and fresh branches. In traditional Chinese medicine, the fruit is used as a medicine. It is cold in nature and bitter in taste. It has the function of clearing heat and purging fire, and mainly treats red eyes, jaundice, hematemesis and heat toxin sores.

According to the present invention, Portulaca oleracea ( Portulaca oleracea ) is a herbaceous plant belonging to the genus Portulaca of the Portulacaceae family ( Portulacaceae ), and is also known as horse lettuce, purslane, purslane and five elements. Purslane is used as medicine in traditional Chinese medicine with the whole plant on the ground. It is sweet and sour, cold in nature and non-toxic.

According to the present invention, the operating procedures and parameter conditions related to fermentation culture fall within the scope of professionalism and routine technology of those who are familiar with the technology.

As used in this article, the terms " Saccharomyces cerevisiae ", " Lactobacillus plantarum " and " Acetobacter aceti " are intended to cover beer that is readily available to those who are familiar with the technology. Yeast, germ lactic acid bacteria and acetic acid bacteria (for example, can be purchased from domestic or foreign depository institutions), or beer yeast and germ isolated and purified from natural sources by using the usual microbial isolation methods in this technology Lactic acid bacteria and acetic acid bacteria strains.

According to the present invention, the medicine can be manufactured into a dosage form suitable for parenterally or orally administration by using techniques well known to those skilled in the art. This includes, but not Limited to: injection (for example, sterile aqueous solution or dispersion), sterile powder, tablet, troche, lozenge (lozenge), pill (pill), capsule (capsule), dispersible powder (dispersible powder) or fine particle (granule), solution, suspension (suspension), emulsion (emulsion), syrup (syrup), elixir (elixir) ), slurry and the like.

The medicine according to the present invention can be administered by a parenteral route selected from the group consisting of: intraperitoneal injection, subcutaneous injection, intramuscular injection (intramuscular injection) and intravenous injection.

The medicine according to the present invention may contain a pharmaceutically acceptable carrier which is widely used in medicine manufacturing technology. For example, the pharmaceutically acceptable carrier may include one or more agents selected from the group consisting of solvents, emulsifiers, suspending agents, decomposers ( decomposer, binding agent, excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative , Lubricant (lubricant), absorption delaying agent (absorption delaying agent), liposome (liposome) and the like. The selection and quantity of these reagents fall within the scope of professionalism and routine techniques of those who are familiar with this technique.

According to the present invention, the pharmaceutically acceptable carrier includes a solvent selected from the group consisting of water, normal saline (normal saline), phosphate buffered saline (PBS), Sugar-containing solutions, aqueous solutions containing alcohol, and combinations thereof.

According to the present invention, a food product can be used as a food additive, which is added during the preparation of raw materials by a conventional method, or added during the preparation of food, and is formulated with any edible material for supply Food products consumed by humans and non-human animals.

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

Example 1. Preparation of plant ferment

First, mix the Chinese gardenia ( Gardenia jasminoides ), Portulaca oleracea ( Portulaca oleracea ), and water in a volume ratio of 1~5:1:40~60, and sterilize and extract them at 50℃~100℃ for 0.5~ In 3 hours, a plant extract was obtained. The plant extract is cooled to room temperature for subsequent three-stage fermentation. The three-stage fermentation system is to sequentially inoculate plant extracts with 0.01~0.5% (v/v) Saccharomyces cerevisiae BCRC 20271 and 0.01~0.25% (v/v) Lactobacillus plantarum TCI028 (BCRC) 910805) Fermentation for 1~5 days; 1~20%(v/v) Acetobacter aceti BCRC 11688 (all the above strains were purchased from Food Industry Research and Development Institute (FIRDI) in Taiwan Fermentation at the Biosource Collection and Research Center (BCRC) for 3 to 8 days. After that, it was concentrated under reduced pressure at 45°C to 70°C, and filtered with a mesh of 200 to 400 mesh. Then, 40-70% isomalto-oligosaccharide is added to adjust the specifications and sterilized to obtain a plant fermented product.

Example 2. Detection of total flavonoids in plant fermented products

The experimental procedure for the detection of total flavonoids content is as follows: the content of total flavonoids is measured with rutin (ChromaDex ASB-00018440) equivalent as the expression of the relative content of total flavonoids. The preparation materials include 10% aluminum nitrat (aqueous solution) (Alfa Aesar 12360), 5% sodium nitrite (aqueous solution) (Sigma 31443), 4% sodium hydroxide (aqueous solution) (Macron 7708-10) and 200μg/mL Rutin (methanol solution).

Take the above Rutin standard solution 0, 200μL, 400μL, 600μL, 800μL, 1000μL and 1200μL respectively into the test tube, respectively add 1200μL, 1000μL, 800μL, 600μL, 400μL, 200μL and 0μL of water, shake and mix evenly; take 200μL Rutin solutions of various concentrations; respectively add 200μL of 5% sodium citrate, mix well and let stand for 6 minutes; add 200μL of 10% aluminum nitrate, mix well and let stand for 6 minutes; then add 2mL of 4% sodium hydroxide After mixing uniformly, add 1.4mL H ₂ O and mix evenly; take 200 μL of the above reaction solution in a 96-well reaction plate, detect the absorbance at 500 nm with a spectrophotometer, and draw a standard curve.

The plant fermented product obtained in Example 1 was used as the experimental group, and the plant extract was used as the comparative group. After proper dilution of the experimental group or comparison group, take 200μL of the sample of the experimental group or comparison group and place it in a test tube; add 200μL of 5% sodium citrate, mix well and let stand for 6 minutes; add 200μL of 10% aluminum nitrate, mix well Then let it stand for 6 minutes; add 2mL 4% sodium hydroxide and mix well, then add 1.4mL H ₂ O and mix well; take 200 μL of the above reaction solution in a 96-well reaction plate, and detect the absorbance at 500 nm with a spectrophotometer. The results of total flavonoid content are shown in Figure 1.

Figure 1 is a data diagram of the total flavonoid content of the fermented plant of the present invention. It can be seen from Figure 1 that compared with the comparison group, the total flavonoid content of the experimental group has a significant increase (up by 1.4 times). The results of this example show that the plant fermented product of the present invention releases a large amount of total flavonoids.

Example 3. Evaluation of the effectiveness of plant fermentation products on cardiovascular health care 3.1 The effect of plant fermentation products in regulating the expression of genes related to atherosclerosis

First, human monocytic cells (human monocytic cell) 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-mercaptoethanol), 100units/mL penicillin (penicillin) and 100μg/mL streptomycin (streptomycin). Add 2 mL of culture medium to each well of the 6-well culture plate so that each well has 1.5 x 10 ⁵ THP-1 cells. Next, a differentiation medium with 500 nM phorbol-12-myristate 13-acetate (PMA) was added and cultured for 48 hours. After that, fresh medium was replaced and cultured for another 48 hours.

In addition, a single plant fermented product (including a single purslane fermented product and a single fermented cocklebur seed) was prepared as the analysis sample of the comparison group, and the preparation method was carried out according to the process of Example 1 above. The difference is: The unilateral purslane and purslane replace the combination of purslane and purslane.

After that, the THP-1 cells were divided into 5 groups, including 1 experimental group, 2 comparison groups (ie, comparison group 1 and comparison group 2), 1 LPS (Lipopolysaccharides) group and 1 control group. Add 2% (v/v) of the plant fermentation product according to Example 1 above to the cells of the experimental group, and add 2% (v/v) of the purslane fermentation product to the cells of the comparative group 1, adding 2% (v/v) The fermented zizi was added to the cells of the comparison group 2, and 100ng/mL lipopolysaccharides (LPS) was added to the cells of the LPS group to induce cell inflammation, which is related to vascular dysfunction (vascular dysfunction) related. As for the cells in the control group, no treatment was done. Then, each group of cells was cultured in an incubator for 24 hours, and then the cell cultures of each group were collected and used for gene expression analysis.

In this embodiment, the genes used to analyze atherosclerosis include the CD36 gene and the ATP binding cassette subfamily A member 1 (ATP binding cassette subfamily A member 1, ABCA1 ) gene.

The RNA extraction kit (Genemark) was used to extract RNA from the cell cultures obtained above. Take 2,000ng of each group of RNA thus obtained and reverse transcribed the extracted RNA into cDNA with SuperScript ^® III reverse transcriptase (Invitrogen). Next, use cDNA as a template and use the primer pairs used to amplify the target gene, including CD36 , ABCA1 and ACTB (as internal controls). Their nucleotide sequences are shown in Table 1 below, in the StepOne Plus real-time PCR system (ABI) uses the KAPA CYBR FAST qPCR kit (2x) (KAPA Biosystems) to perform quantitative real-time PCR to amplify and quantify the target gene. The melting curve of the PCR product is confirmed during the quantitative real-time PCR reaction.

，並利用ACTB基因(作為內部對照組)及基準基因的循環閾值及藉由標準差來計算相對倍數變化，其中△Ct=Ct_{目標基因/基準基因}-Ct_ACTB，△△Ct=△Ct_目標基因-△Ct_基準基因，

。以對照組的標的基因表現量作為1的比較基準。各組之間的統計學顯著差異是藉由單尾史徒登氏t-檢定來決定。本實施例的結果顯示於圖2。 The relative expression of the target gene is derived from the equation

, And use ACTB gene (as the internal control group) and the cycle threshold of the reference gene and use the standard deviation to calculate the relative fold change, where △Ct=Ct _{target gene/reference gene-} Ct _ACTB , △△Ct=△Ct _{target gene} -△Ct _{reference gene} ,

. The target gene expression level of the control group was used as the comparison benchmark of 1. The statistically significant difference between the groups was determined by the one-tailed Studen's t-test. The results of this example are shown in Figure 2.

Figure 2 is a data diagram showing the efficacy of the plant fermented product of the present invention in regulating the expression of genes related to atherosclerosis (including CD36 gene and ABCA1 gene). It can be seen from Figure 2 that in terms of CD36 gene, compared with the control group, LPS group, comparison group 1 and comparison group 2, the gene fold change of the experimental group has a significant reduction, which is about 70% lower than that of the control group. %. As far as the ABCA1 gene is concerned, compared with the control group, the LPS group, the comparison group 1 and the comparison group 2, the gene fold change of the experimental group has a significant increase, which is about 100% higher than that of the control group. The results of this experiment show that the plant fermented product of the present invention can regulate the expression of genes related to atherosclerosis (including CD36 gene and ABCA1 gene), avoid the formation of foam cells, reduce the risk of atherosclerosis, and achieve the effect of cardiovascular health care. .

3.2 The effect of plant fermentation products in regulating the expression of genes related to cardiovascular health

First, human monocytic cells (human monocytic cell) 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-mercaptoethanol), 100units/mL penicillin (penicillin) and 100μg/mL streptomycin (streptomycin). Add 2 mL of culture medium to each well of the 6-well culture plate so that each well has 1.5 x 10 ⁵ THP-1 cells. Next, a differentiation medium with 500 nM phorbol-12-myristate 13-acetate (PMA) was added and cultured for 48 hours. After that, fresh medium was replaced and cultured for another 48 hours.

In addition, a single plant fermented product (including a single purslane fermented product and a single fermented cocklebur seed) was prepared as the analysis sample of the comparison group, and the preparation method was carried out according to the process of Example 1 above. The difference is: The unilateral purslane and purslane replace the combination of purslane and purslane.

After that, the THP-1 cells were divided into 4 groups, including 1 experimental group, 2 comparison groups (ie, comparison group 1 and comparison group 2), and 1 control group. 0.5% (v/v) of the plant fermentation product according to Example 1 above was added to the cells of the experimental group, 0.5% (v/v) of the purslane fermentation product was added to the cells of the comparative group 1, and 0.5 %(v/v) the fermented sesame seeds were added to the cells of the comparative group 2. As for the cells in the control group, no treatment was done. Then, each group of cells was cultured in an incubator for 24 hours, and then the cells were treated with a cell lysis buffer, and then the cell cultures of each group were collected and used for gene expression analysis.

In this embodiment, the genes used to analyze cardiovascular health care include protein C ( PROC ) gene, von Willebrand factor ( VWF ) gene, F3 gene, serine protease inhibitor Family E member 1 (serine protease inhibitor family E member 1, SERPINE1 ) gene, platelet derived growth factor C ( PDGFC ) gene, fibroblast growth factor 2 ( FGF2 ) gene, similar Insulin-like growth factor 2 mRNA-binding protein 3 (Insulin-like growth factor 2 mRNA-binding protein 3, IGF2BP3 ) gene, insulin like growth factor 1 receptor ( IGF1R ) gene, interleukin 8 (Interleukin -8, IL8 ) gene, vascular cell adhesion molecule 1 ( VCAM1 ) gene and apoptosis protease 8 (caspase 8, CASP8 ) gene.

The RNA extraction kit (Genemark) was used to extract RNA from the cell cultures obtained above. Take 2,000ng of each group of RNA thus obtained and reverse transcribed the extracted RNA into cDNA with SuperScript ^® III reverse transcriptase (Invitrogen). Next, use cDNA as a template, and use primer pairs used to amplify target genes, including PROC, VWF, F3, SERPINE1, PDGFC, FGF2, IGF2BP3, IGF1R, IL8, VCAM1, CASP8, and ACTB (as an internal control group), Their nucleotide sequences are shown in Table 2 below. The KAPA CYBR FAST qPCR kit (2x) (KAPA Biosystems) is used in the StepOne Plus real-time PCR system (ABI) to perform quantitative real-time PCR to amplify the target gene And quantitative. The melting curve of the PCR product is confirmed during the quantitative real-time PCR reaction.

，並利用ACTB基因(作為內部對照組)及基準基因的循環閾值及藉由標準差來計算相對倍數變化，其中△Ct=Ct_{目標基因/基準基因}-Ct_ACTB，△△Ct=△Ct_目標基因-△Ct_基準基因，

。以對照組的標的基因表現量作為1的比較基準。各組之間的統計學顯著差異是藉由單尾史徒登氏t-檢定來決定。本實施例的結果顯示於圖3至圖6。 The relative expression of the target gene is derived from the equation

, And use ACTB gene (as the internal control group) and the cycle threshold of the reference gene and use the standard deviation to calculate the relative fold change, where △Ct=Ct _{target gene/reference gene-} Ct _ACTB , △△Ct=△Ct _{target gene} -△Ct _{reference gene} ,

. The target gene expression level of the control group was used as the comparison benchmark of 1. The statistically significant difference between the groups was determined by the one-tailed Studen's t-test. The results of this example are shown in Figures 3 to 6.

Figures 3 to 6 are the effects of the plant fermented product of the present invention in regulating the performance of genes related to cardiovascular health care (including PROC, VWF, F3, SERPINE1, PDGFC, FGF2, IGF2BP3, IGF1R, IL8, VCAM1 , and CASP8 genes) The data graph. It can be seen from Figure 3 that in terms of PROC gene, compared with the control group, comparison group 1 and comparison group 2, the relative gene expression of the experimental group has a significant increase, which is about 320% higher than that of the control group; As far as the VWF gene is concerned, compared with the control group, comparison group 1 and comparison group 2, the relative gene expression level of the experimental group has a significant decrease, which is about 70% lower than that of the control group; as far as the F3 gene is concerned Compared with the control group, comparison group 1 and comparison group 2, the relative gene expression level of the experimental group has a significant decrease, which is about 75% lower than that of the control group; for the SERPINE1 gene, compared with the control group, Compared with the comparison group 1 and the comparison group 2, the relative gene expression of the experimental group has a significant reduction, which is about 20% lower than that of the control group.

As can be seen from Figure 4, as far as the PDGFC gene is concerned, compared with the control group, the comparison group 1 and the comparison group 2, the relative gene expression of the experimental group has a significant reduction, which is about 75% lower than that of the control group; As far as the FGF2 gene is concerned, compared with the control group, comparison group 1 and comparison group 2, the relative gene expression of the experimental group has a significant decrease, which is about 30% lower than that of the control group; as for the IGF2BP3 gene Compared with the control group, the comparison group 1 and the comparison group 2, the relative gene expression of the experimental group has a significant reduction, which is about 60% lower than that of the control group; in terms of the IGF1R gene, compared with the control group, Compared with the comparison group 1 and the comparison group 2, the relative gene expression of the experimental group has a significant reduction, which is about 45% lower than that of the control group.

As can be seen from Figure 5, as far as the IL8 gene is concerned, compared with the control group, the comparison group 1 and the comparison group 2, the relative gene expression of the experimental group has a significant reduction, which is about 45% lower than that of the control group; As far as the VCAM1 gene is concerned, compared with the control group, comparison group 1 and comparison group 2, the relative gene expression of the experimental group has a significant reduction, which is about 60% lower than that of the control group.

As can be seen from Figure 6, in terms of CASP8 gene, compared with the control group, comparison group 1 and comparison group 2, the relative gene expression of the experimental group has a significant reduction, which is about 60% lower than that of the control group.

The results of this experiment show that the plant fermented product of the present invention can inhibit thrombosis by enhancing the expression of PROC gene; inhibiting the expression of VWF , F3 , and SERPINE1 genes, reducing thrombosis; by inhibiting PDGFC , FGF2 , IGF2BP3 , and IGF1R genes It can inhibit the vasoconstriction caused by the proliferation of vascular cells and regulate blood pressure; by inhibiting the expression of IL8 , VCAM1 , and CASP8 genes, it can reduce blood coagulation.

In summary, the plant fermented product of the present invention can regulate CD36 gene, ABCA1 gene, PROC gene, VWF gene, F3 gene, SERPINE1 gene, PDGFC gene, FGF2 gene, IGF2BP3 gene, IGF1R gene, IL8 gene, VCAM1 gene and The expression of CASP8 gene regulates the whole heart meridian in the five elements meridian, regulates the expression of vasodilation and atherosclerosis genes, reduces thrombosis, regulates angiogenesis genes, relaxes blood pressure, and reduces vascular inflammation genes, reduces coagulation response, and reaches the cardiovascular system health benefits.

The above description is only illustrative, not restrictive. Any equivalent modification or alteration that does not deviate from the spirit and scope of the present invention shall be included in the scope of the appended patent application.

<110> Dajiang Biomedical Co., Ltd.

<120> Plant fermentation products and their uses

<130> 107B0608-I1

<160> 28

<170> PatentIn version 3.5

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<213> Artificial sequence

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<213> Artificial sequence

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<213> Artificial sequence

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<213> Artificial sequence

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<213> Artificial sequence

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<213> Artificial sequence

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<213> Artificial sequence

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<213> Artificial sequence

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Claims (6)

A plant fermented product is prepared by a method including the following steps: (a) Extracting a combination of gardenia jasminoides and Portulaca oleracea with water to obtain a plant extract Wherein the plant extract is extracted at 50°C to 100°C for 0.5 to 3 hours; and (b) the plant extract is combined with Saccharomyces cerevisiae , Lactobacillus plantarum , and acetic acid Bacteria ( Acetobacter aceti ) are fermented sequentially to obtain the plant fermented product, wherein the fermentation time of the beer 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; the beer The concentration of yeast is between 0.01~0.5%(v/v), the concentration of the germ lactic acid bacteria is between 0.01~0.25%(v/v), and the concentration of the acetic acid bacteria is between 1~20%(v/v) ; The volume ratio of the zizi, the purslane and the water is between 1~5:1:40~60. The effective concentration of the plant fermented product described in item 1 of the scope of patent application is at least 2% (v/v). A plant fermentation product is used to prepare a regulatory CD36 gene, ATP binding cassette subfamily A member 1 (ATP binding cassette subfamily A member 1, ABCA1 ) gene, protein C ( PROC ) gene, and von Willebrand coagulation factor (von Willebrand). factor, VWF ) gene, F3 gene, serine protease inhibitor family E member 1 (serine protease inhibitor family E member 1, SERPINE1 ) gene, platelet derived growth factor C ( PDGFC ) gene, fibroblast Cell growth factor 2 (fibroblast growth factor 2, FGF2 ) gene, Insulin-like growth factor 2 mRNA-binding protein 3 ( IGF2BP3 ) gene, insulin-like growth factor 1 receptor (insulin-like growth factor 2 mRNA-binding protein 3, IGF2BP3 ) gene growth factor 1 receptor, IGF1R ) gene, interleukin-8 (Interleukin-8, IL8 ) gene, vascular cell adhesion molecule 1 ( VCAM1 ) gene and apoptosis protease 8 (caspase 8, CASP8 ) gene The use of expressive composition, wherein the plant fermented product is prepared by a method including the following steps: (a) Extracting a composition made of Gardenia jasminoides and Portulaca oleracea with water To obtain a plant extract, wherein the plant extract is extracted at 50°C to 100°C for 0.5 to 3 hours; and (b) the plant extract and Saccharomyces cerevisiae , Lactobacillus plantarum and Acetobacter aceti are fermented in sequence to obtain the plant fermentation product, wherein the fermentation time of the brewer's yeast and the embryo lactic acid bacteria is 1 to 5 days, and the fermentation time of the acetic acid bacteria 3 to 8 days; the concentration of the brewer's yeast is between 0.01~0.5% (v/v), the concentration of the germ lactic acid bacteria is between 0.01~0.25% (v/v), and the concentration of the acetic acid bacteria is between 1 ~20%(v/v); the volume ratio of the zizi, the purslane and the water is between 1~5:1 : 40~60. A plant fermented product is used to prepare a composition that promotes vasodilation and angiogenesis, alleviates atherosclerosis and vascular inflammation, reduces thrombosis, relaxes blood pressure, and reduces coagulation reaction, wherein the plant fermented product includes It is prepared by the following steps: (a) Extracting a combination of Gardenia jasminoides and Portulaca oleracea with water to obtain a plant extract, wherein the plant extract is 50% It is obtained by extracting at ℃~100℃ for 0.5~3 hours; and (b) fermenting the plant extract with Saccharomyces cerevisiae , Lactobacillus plantarum and Acetobacter aceti in sequence, The plant fermentation product is obtained, 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; the concentration of the brewer's yeast is 0.01~0.5% (v/v), the concentration of the germ lactic acid bacteria is between 0.01~0.25%(v/v), and the concentration of the acetic acid bacteria is between 1~20%(v/v); the zi, the purslane and The volume ratio of the water is between 1~5:1:40~60; the plant fermented product is concentrated under reduced pressure at 45℃~70℃, filtered with a 400 mesh screen, and then added 40~ 70% isomalt-oligosaccharides are prepared after adjusting the specifications and sterilizing. For the use described in item 3 or item 4 of the scope of patent application, the effective concentration of the plant fermented product is at least 2% (v/v). Such as the use described in item 3 or item 4 of the scope of patent application, wherein the composition is in the form of a medicine or a food product.

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