Chrysanthemum fermented product as well as preparation method and application thereof
By drying, crushing, sieving and soaking the chrysanthemum flowers, and using probiotics to ferment them, chrysanthemum fermentation products are prepared, which solves the problem of low abundance of active ingredients of existing chrysanthemum extracts and significantly improves its blood pressure-lowering effect.
Patent Information
- Application Number
- CN202510023861.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-07
- Publication Date
- 2025-04-15
AI Technical Summary
When preparing the existing chrysanthemum extract, the active ingredients are low, resulting in poor blood pressure lowering effect.
After the chrysanthemum flowers are dried, crushed, sieved and soaked, probiotics such as lactic acid bacteria and yeast are fermented to prepare chrysanthemum fermented products.
It improves the abundance of active ingredients in chrysanthemum fermented products, increases the types and content of active substances for lowering blood pressure, and improves the blood pressure-lowering effect of chrysanthemum extract.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of natural product extraction and activity, and specifically relates to a chrysanthemum fermentation product and a preparation method and application thereof. Background Art
[0002] Chrysanthemum is the dried inflorescence of the plant chrysanthemum of the Asteraceae family. It was first seen in Shennong's Herbal Classic. It tastes sweet and bitter, is slightly cold, and enters the lung and liver meridians. It has traditional effects such as clearing away heat and detoxifying, calming the liver and improving eyesight. Clinically, chrysanthemum is often used in combination with other liver-calming and yang-inhibiting drugs to treat headaches, dizziness, red and swollen eyes, and other related symptoms caused by hypertension.
[0003] Modern pharmacological studies have shown that chrysanthemum extract can reduce the blood pressure of rats with renal hypertension and improve their myocardial hypertrophy. Chrysanthemum total flavonoids have a hypotensive effect on spontaneously hypertensive rats. The preparation of chrysanthemum extract requires the use of a method for extracting effective substances in traditional Chinese medicine. At present, chrysanthemum extract is mostly prepared by aqueous phase boiling and ethanol extraction. The above two methods are simple to operate, but the abundance of effective ingredients is low, which reduces the hypotensive effect of chrysanthemum extract. Summary of the invention
[0004] In order to solve the problem that the existing chrysanthemum extract has a low abundance of effective ingredients during preparation, thereby reducing the blood pressure lowering effect of the chrysanthemum extract, the present invention adopts the following technical solution.
[0005] The present invention provides a method for preparing a chrysanthemum fermented product, comprising the following steps:
[0006] The chrysanthemum flowers are dried, crushed and sieved to obtain chrysanthemum powder.
[0007] A solvent is added to the chrysanthemum powder for soaking to obtain a mixture of the chrysanthemum and the soaking solution.
[0008] After the probiotics are activated, they are inoculated into the mixture of the chrysanthemum and the soaking solution for fermentation and culture. The probiotics include any one or more of lactic acid bacteria and yeast.
[0009] After the fermentation is finished, the fermentation liquid is filtered and the chrysanthemum is removed to obtain the chrysanthemum fermented product.
[0010] The present invention provides a method for preparing a chrysanthemum fermented product, which uses chrysanthemum flowers as materials, and then undergoes processes such as drying, crushing, sieving and soaking, and uses the probiotics for fermentation, which can improve the abundance of effective ingredients in the fermented product using chrysanthemum as a substrate compared to chrysanthemum extract. The chrysanthemum fermented product prepared by the preparation method of the chrysanthemum fermented product provided by the present invention has more types of antihypertensive active substances, and has more small molecular substances with antihypertensive activity such as polysaccharides, small peptides and other substances, thereby improving the antihypertensive effect of the chrysanthemum extract.
[0011] Preferably, the inoculation amount of the probiotics is 0.5%wt to 1.5%wt.
[0012] Preferably, the probiotics include lactic acid bacteria and yeast, and the lactic acid bacteria are Lactobacillus.
[0013] The inoculation volume ratio of the lactobacillus to the yeast is 1:1.
[0014] Preferably, the lactobacillus is Lactobacillus plantarum; and the yeast is Saccharomyces cerevisiae.
[0015] Preferably, the activation and inoculation methods of the Lactobacillus plantarum and the Saccharomyces cerevisiae are as follows:
[0016] The plant lactobacillus is cultured in an MRS medium at 37° C. for 12 hours to obtain activated plant lactobacillus.
[0017] The brewer's yeast is cultured in a YPD medium at 30° C. for 32 hours to obtain activated brewer's yeast.
[0018] After the activated Lactobacillus plantarum and the activated Saccharomyces cerevisiae grow to the logarithmic growth phase, they are inoculated into the mixture of the chrysanthemum and the soaking solution at an inoculation amount of 0.5%wt to 1.5%wt respectively.
[0019] Preferably, the fermentation culture conditions are: 25°C to 35°C, and fermentation for 15 days to 30 days. Such fermentation culture conditions can provide the optimal fermentation conditions for Lactobacillus plantarum and Saccharomyces cerevisiae.
[0020] Preferably, the material-liquid ratio of the chrysanthemum powder to the added solvent is 0.2 g / mL to 0.4 g / mL.
[0021] The solvent is water.
[0022] The present invention also provides a chrysanthemum fermented product prepared by the preparation method of the chrysanthemum fermented product. The chrysanthemum fermented product provided by the present invention has a high abundance of effective ingredients, increases the abundance of antihypertensive active substances in the chrysanthemum extract, has more small molecule substances with antihypertensive activity such as polysaccharides, small peptides and the like, and is conducive to further improving the antihypertensive effect.
[0023] The present invention also provides application of the chrysanthemum fermented product in improving the blood pressure lowering effect of the chrysanthemum extract.
[0024] Preferably, the chrysanthemum fermented product is used to increase the abundance of antihypertensive active substances in the chrysanthemum extract.
[0025] Compared with the prior art, the present invention has the following beneficial effects:
[0026] 1. The present invention provides a chrysanthemum fermented product and a preparation method thereof. The chrysanthemum fermented product and the preparation method thereof provided by the present invention use chrysanthemum flowers as materials, and then go through processes such as drying, crushing, sieving and soaking, and use the probiotics for fermentation, which can improve the abundance of effective ingredients in fermented products using chrysanthemum as a substrate compared to chrysanthemum extracts. The chrysanthemum fermented product prepared by the preparation method of the chrysanthemum fermented product provided by the present invention has more types of antihypertensive active substances, and has more small molecular substances with antihypertensive activity such as polysaccharides, small peptides and other substances, thereby improving the antihypertensive effect of chrysanthemum extracts.
[0027] 2. The present invention studies the hypotensive and antioxidant activities of chrysanthemum flowers during fermentation with probiotics, and finds that the hypotensive ability of chrysanthemums is increased by fermentation with probiotics. The present invention provides a fermented product with chrysanthemums as a substrate that has both hypotensive and antioxidant effects.
[0028] 3. The preparation method of the chrysanthemum fermented product provided by the present invention utilizes the growth of plant lactobacillus to change the acidity and alkalinity of the solvent, and utilizes the growth of saccharomyces cerevisiae to change the polarity of the solvent. Through these complex and continuous changes, it is beneficial to the leaching of more active substances. At the same time, in addition to changing the acidity and alkalinity and polarity of the solvent, the growth of microorganisms can further decompose macromolecular substances such as sugars and proteins in chrysanthemums, and produce more small molecular substances with antihypertensive activity such as polysaccharides and small peptides, which is beneficial to further improve the antihypertensive effect.
[0029] 4. The fermentation method of using safe and probiotic strains such as Lactobacillus plantarum and Saccharomyces cerevisiae as a new method of extracting limited substances from traditional Chinese medicine can greatly improve the efficacy of traditional Chinese medicine.
[0030] 5. The preparation method of a chrysanthemum fermented product provided by the present invention only needs to be fermented in a simple fermentation device without the need for a sterilization process. The antihypertensive effect of chrysanthemum can be greatly improved through the fermentation of Lactobacillus plantarum and Saccharomyces cerevisiae. DETAILED DESCRIPTION
[0031] The present invention is described in detail below in conjunction with specific examples, but it should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.
[0032] Example 1
[0033] A method for preparing a chrysanthemum fermented product comprises the following steps:
[0034] 1. Bacteria activation
[0035] The Lactobacillus plantarum was cultured in MRS medium at 37° C. for 12 hours to obtain activated Lactobacillus plantarum, wherein the Lactobacillus plantarum has a preservation number of CICC 20982.
[0036] The brewer's yeast was cultured in a YPD medium at 30° C. for 32 hours to obtain activated Lactobacillus plantarum. The brewer's yeast has a preservation number of CICC 1001.
[0037] Lactobacillus plantarum and Saccharomyces cerevisiae were purchased from China Industrial Microbiological Culture Collection Center.
[0038] 2. Preparation of chrysanthemum aqueous solution
[0039] 200 g of chrysanthemum flowers were dried at 65° C. for 1 h, crushed and passed through a 10-mesh sieve to obtain chrysanthemum powder. The chrysanthemum powder was soaked in 1000 mL of water that was boiled and then cooled to room temperature, and the obtained mixture of chrysanthemum and soaking solution was the chrysanthemum aqueous solution.
[0040] The chrysanthemum aqueous solution has a mass ratio of 20%.
[0041] Among them, the chrysanthemum flowers were purchased from Hangbaiju purchased from Beijing Tong Ren Tang.
[0042] It should be noted that chrysanthemum pollen can be inoculated after mixing with water.
[0043] 3. Fermentation culture
[0044] After the activated Lactobacillus plantarum and the activated Saccharomyces cerevisiae have reached the logarithmic growth phase, the activated Lactobacillus plantarum and the activated Saccharomyces cerevisiae are inoculated into the chrysanthemum aqueous solution in an inoculation amount of 1%wt in a special sealed fermentation barrel and fermented and cultured at 30°C for 15 days.
[0045] The inoculation volume ratio of the activated Lactobacillus plantarum and the activated Saccharomyces cerevisiae is 1:1.
[0046] After the fermentation is completed, the fermentation liquid is centrifuged at 4°C and 4000 rpm / min to obtain the supernatant, which is freeze-dried to obtain the freeze-dried powder, which is the chrysanthemum fermentation product.
[0047] Comparative Example 1
[0048] A method for preparing a chrysanthemum aqueous solution comprises the following steps:
[0049] 200g of chrysanthemum was dried at 65℃ for 1h, crushed and passed through a 10-mesh sieve to obtain chrysanthemum powder. The chrysanthemum powder was soaked in 1000mL of water that was boiled and then cooled to room temperature. After soaking at 30℃ for 15 days, the obtained soaking solution was the chrysanthemum aqueous solution.
[0050] The chrysanthemum aqueous solution has a mass ratio of 20%.
[0051] Among them, the chrysanthemum source is Hangzhou white chrysanthemum purchased from Beijing Tong Ren Tang.
[0052] Comparative Example 2
[0053] A method for preparing a chrysanthemum alcohol solution comprises the following steps:
[0054] 200g of chrysanthemum was dried at 65℃ for 1h, crushed and passed through a 10-mesh sieve to obtain chrysanthemum powder. The chrysanthemum powder was soaked in 1000mL of liquor. After soaking at 30℃ for 15d, the obtained soaking solution was chrysanthemum alcohol solution.
[0055] The mass ratio of the chrysanthemum alcohol solution is 20%.
[0056] Among them, the chrysanthemum source is Hangzhou white chrysanthemum purchased from Beijing Tong Ren Tang.
[0057] The source of the liquor is 4L bottle of Beida Warehouse 52% alcohol liquor.
[0058] In order to illustrate the effect of the preparation method of a chrysanthemum fermented product provided by the present invention, we take the chrysanthemum fermented product prepared in Example 1, the chrysanthemum aqueous solution prepared in Comparative Example 1, and the chrysanthemum alcohol solution prepared in Comparative Example 2 as examples, take the preparation method of a chrysanthemum fermented product provided in Example 1 as the fermentation group, the preparation method of a chrysanthemum aqueous solution provided in Comparative Example 1 as the control group 1, and the preparation method of a chrysanthemum alcohol solution provided in Comparative Example 2 as the control group 2, and the in vitro and in vivo antihypertensive activity and antioxidant activity of the chrysanthemum fermented product obtained in Example 1, the chrysanthemum aqueous solution obtained in Comparative Example 1, and the chrysanthemum alcohol solution obtained in Comparative Example 2 are studied as follows:
[0059] 1. Experimental Methods
[0060] 1. In vitro ACE activity detection
[0061] Required reagent samples: sodium tetraborate, sodium chloride, boric acid, maleuryl histidyl leucine, ACE enzyme, sodium hydroxide, hydrochloric acid, ethyl acetate, quinoline, benzenesulfonyl chloride, anhydrous ethanol were purchased from Shanghai Shenggong Biotechnology Co., Ltd.
[0062] 1.1 Drug preparation:
[0063] 0.1mol / L boric acid buffer, also known as BBS buffer, is prepared as follows:
[0064] Solution 1: 1.9068g sodium tetraborate, 3.5064g sodium chloride, add water to make up to 200mL.
[0065] Solution 2: 1.2366g boric acid, 3.5064g sodium chloride, add water to make up to 200mL.
[0066] Mix solution 1 and solution 2, adjust the pH to 8.3, and store at 4°C.
[0067] The preparation method of 5mmol / L hippocampus histidyl leucine solution is as follows: dissolve 25mg hippocampus histidyl leucine in 11.64mL of the above BBS buffer and store at -20℃. Dispense into 2mL centrifuge tubes, 500μL per tube.
[0068] The preparation method of 0.1 UN / mL ACE solution is as follows: dissolve 0.1 UN ACE enzyme in 1 mL BBS buffer and store at -80°C. Dispense in 0.2 mL centrifuge tubes, 50 μL per tube.
[0069] The preparation method of 1M HCL is: 8.5mL of 36.5% HCL by volume is added with water to make 100mL. During the preparation process, add water first and then add HCL.
[0070] 1.2 Experimental steps:
[0071] ① Treatment of samples after fermentation: centrifuge 1 L of the fermented chrysanthemum product sample prepared in Example 1 at 4° C. and 4000 rpm / min; take the supernatant after centrifugation, and freeze-dry it to obtain a lyophilized powder, which is the sample to be tested.
[0072] ② The treatment method of the chrysanthemum aqueous solution obtained in Comparative Example 1 and the chrysanthemum alcohol solution obtained in Comparative Example 2 is the same.
[0073] ③Take three groups of 10mL centrifuge tubes, one for the blank group, one for the sample group, and one for the control group. The corresponding centrifuge tubes are called blank tubes, sample tubes, and control tubes, respectively.
[0074] ④ Mix the sample to be tested with HHL: Take 50 μL of 5mmol / L HHL solution and mix it with 20 μL of sample solution, and make sample control (BBS instead of the sample to be tested) and blank control (use 100 μL HCL solution to inactivate ACE enzyme). All the above groups react at 37℃ for 5 minutes.
[0075] ⑤ Add ACE reactant: Add 10 μL of 0.1U / mL ACE solution to the sample tube and the control tube, mix well, do not add to the blank tube, and react at 37°C for 30 minutes.
[0076] ⑥Terminate the reaction and generate hippuric acid: ACE hydrolyzes HHL to generate hippuric acid. Add 100 μL HCl solution to terminate the reaction. Do not add to the blank tube.
[0077] ⑦ Add 20 μL of the sample to be tested to the control tube, and dilute the total volume of each tube to 0.5 mL with borate buffer solution, including 320 μL each to the sample tube and blank tube, and 300 μL to the control tube.
[0078] ⑧ Add the developer: add 0.6 mL of quinoline to each tube of reaction solution in the dark, vortex mix for 10 seconds, then add 0.2 mL of benzenesulfonyl chloride, vortex mix for 20 seconds, and react at 30°C in the dark for 30 minutes.
[0079] ⑨ Add ethanol: Add 3.7 mL of anhydrous ethanol to each tube of reaction solution, mix well, and continue to place in the dark for 30 minutes.
[0080] ⑩ Determine absorbance: Take 0.2 mL of the reaction solution and add it to a 96-well flat-bottom culture plate. Use an ELISA reader to measure the absorbance at a wavelength of 492 nm.
[0081] Calculation formula:
[0082]
[0083] Where: A a -Absorbance in the presence of both ACE and sample (sample);
[0084] A b -Absorbance without sample participation in the reaction (control);
[0085] A c -Absorbance without ACE enzyme involved in the reaction (blank);
[0086] The unit of ACE inhibition rate is %.
[0087] It should be noted that each sample corresponds to a control and a blank. Here, "each sample" refers to the chrysanthemum fermented product with antihypertensive activity obtained in Example 1, the chrysanthemum aqueous solution obtained in Comparative Example 1, and the chrysanthemum alcohol solution obtained in Comparative Example 2.
[0088] 2. In vitro antioxidant activity detection
[0089] DPPH solution is ready for use: accurately weigh 0.004 g DPPH and dilute to 100 mL in a brown volumetric flask with anhydrous ethanol.
[0090] Take 2mL of the sample to be tested and add it to a test tube, add 2mL of 0.01mol / L DPPH free radical anhydrous ethanol solution, and react for 30 minutes in the dark, then measure the absorbance value Ai at 517nm; replace the DPPH free radical anhydrous ethanol solution with anhydrous ethanol and measure the absorbance value Aj; the reaction uses 2mL of anhydrous ethanol added to 2mL of DPPH free radical anhydrous ethanol solution as a reference, and the absorbance value is A0. All measurements are performed in triplicate and the average value is taken.
[0091] DPPH free radical scavenging rate calculation formula:
[0092]
[0093] Where: Ai- is the absorbance value of sample and DPPH free radical at 517nm;
[0094] Aj-absorbance value of sample and anhydrous ethanol at 517nm;
[0095] The absorbance value of A0-DPPH free radical and anhydrous ethanol at 517nm.
[0096] 3. Animal experiments
[0097] 3.1 Experimental animals
[0098] The experimental animals were SPF-grade Wistar rats, male, weighing 0.20±0.02kg, provided by Changchun Yisi Experimental Animal Technology Co., Ltd.
[0099] 3.2 Drugs and reagents
[0100] L-NAME was purchased from Shanghai Yuanye Biological Co., Ltd. Captopril was purchased from Shanghai Xudong Haipu Pharmaceutical Co., Ltd.
[0101] 3.3 Main instruments and equipment
[0102] The BP-100A fully automatic non-invasive blood pressure measurement system for mice and rats was purchased from Chengdu Taimeng Software Co., Ltd. The TGL-16G high-speed desktop centrifuge was purchased from Shanghai Anting Scientific Instrument Factory.
[0103] 3.4 Experimental methods
[0104] 3.4.1 Preparation of test drugs
[0105] Freeze-dried products: distilled water was added to the fermentation group, control group 1, and control group 2 to prepare solutions with a concentration of 25 mg / mL for standby use.
[0106] L-NAME solution: Add distilled water to prepare a solution with a content of 1.8 mg / mL and set aside.
[0107] Captopril solution: Grind the captopril tablets into fine powder in a mortar and pestle, add distilled water to prepare a solution with a content of 0.5 mg / mL, and set aside.
[0108] 3.4.2 Animal grouping and drug administration
[0109] Wistar rats were given free access to water and food and were adaptively fed for one week.
[0110] According to the principle of random grouping, Wistar rats after one week of adaptive feeding were divided into a normal rat fermentation group, a control group water extract, and a control group alcohol extract. The dosage was 250 mg / kg, with a total of 3 groups, 9 rats in each group.
[0111] 3.4.3 Experiment on the effect of chrysanthemum extract on blood pressure in hypertensive rats
[0112] ①Establishment of a hypertensive rat model
[0113] Healthy male Wistar rats with a body weight of 200±20g were provided with ordinary feed and distilled water for one week of adaptive feeding. After the adaptive feeding, L-NAME was given by gavage at a dose of 18 mg / kg at 9 am every day to establish the model. The drug induction lasted for 4 weeks, and the blood pressure of the rats was measured once a week.
[0114] After drug induction, blood pressure was measured using a non-invasive blood pressure meter. If the rat's systolic blood pressure was ≥140 mmHg, the model was successfully established.
[0115] ② Animal grouping and drug administration
[0116] Wistar rats with successful hypertension modeling were selected and randomly divided into 1 mg / kg captopril positive control group, hypertensive rat fermentation group, control group water extract, and control group alcohol extract. The dosage was 250 mg / kg, with a total of 4 groups, 9 rats in each group.
[0117] 3.4.4 Measurement of rat tail artery pressure and body weight
[0118] At a fixed time every week, the rat tail artery pressure was measured with the BP-100A non-invasive blood pressure measurement and analysis system when the rats were awake and quiet. The systolic blood pressure, diastolic blood pressure and mean arterial pressure of the rats in the system were recorded, and the measurements were taken three times in a row to obtain the average value. At the same time, the changes in the body weight of the rats in each group before and after treatment were compared at the same time every week.
[0119] ③ Specimen collection
[0120] After the rats were intragastrically administered, 3 mL / 1 kg of 10% chloral hydrate solution was injected intraperitoneally for anesthesia. 5 mL of blood was quickly collected from the rat heart and placed in a blood collection tube without anticoagulant. The blood was centrifuged at 3500 r / min for 10 min at 4°C. The supernatant was separated and stored at -20°C for the determination of physiological and biochemical indicators in rat serum. The rats were killed after blood collection.
[0121] The levels of angiotensin, renin, aldosterone, nitric oxide, endothelial nitric oxide synthase, tumor necrosis factor α, interleukin-6, endothelin, superoxide dismutase, malondialdehyde, total antioxidant capacity, reduced glutathione, and atrial natriuretic peptide in rat serum were determined by Beijing Huaying Biotechnology Institute using kits developed by Beijing Huaying Biotechnology Institute.
[0122] 2. Experimental Results
[0123] 1. In vitro experimental results and discussion
[0124] The data of ACE enzyme activity inhibition detected in vitro showed that the activity of the fermentation group was more than 2 times that of the control group 1 and the control group 2. The data of DPPH clearance detected in vitro showed that the activity of the fermentation group was about 1.5 of that of the control group 1 and the control group 2, as shown in Table 1.
[0125] The fermentation group had a significantly better blood pressure suppressive effect on hypertensive rats than the control group, and showed strong antioxidant activity in vivo.
[0126] Table 1 In vitro biological activity comparison
[0127] Test substances Fermentation Group Control group 1 Control group 2 ACE inhibition rate 82.15％ 32.64％ 39.38％ DPPH clearance 50.66％ 30.13％ 28.84％
[0128] 2. Animal Experiment Results and Discussion
[0129] 2.1 Content of blood pressure lowering active substances
[0130] By detecting the contents of blood pressure-related substances such as IL-6, TNF-α, ANP, AII, ALD, ET-1, Renin, and eNOS in rat serum, the mechanism of action of chrysanthemum extract in rats can be further determined.
[0131] Among them, IL-6: reduces the content of prostacyclin (PGI2) in endothelial cells and increases Ca 2+ The penetration and concentration of chloramphenicol will also cause arterial vasoconstriction and increase in blood pressure.
[0132] TNF-α: Increased levels of TNF-α will increase the Ca+ content in smooth muscle cells, causing vasoconstriction and increased blood pressure.
[0133] ANP promotes the production of IL-6, thereby promoting the secretion of platelet-derived growth factor, cell fibrosis and proliferation of vascular smooth muscle cells, resulting in a decrease in the diameter of the blood vessel wall, an increase in vascular resistance, and ultimately an increase in blood pressure.
[0134] ANP: Atrial natriuretic peptide, which relaxes vascular smooth muscle and promotes renal excretion of sodium and water. When blood pressure rises, it can stimulate atrial muscle cells to release ANP, thereby relaxing blood vessels and reducing peripheral resistance.
[0135] AII: ACE converts AI into active AII, which is a powerful vasoconstrictor. On the one hand, it acts on the receptors on the blood vessel wall to constrict the surrounding arterioles and smooth muscle vessels, causing blood pressure to rise. On the other hand, it stimulates the secretion of ALD and promotes the reabsorption of Na+ and K+ by the kidneys, causing blood pressure to rise.
[0136] ALD: Aldosterone is a steroid hormone secreted by the adrenal cortex. When the AII level decreases, the vasoconstrictor activity decreases, leading to reduced ALD secretion and lower blood pressure.
[0137] ET-1: A subtype of endothelin, a vasoconstrictor factor. It is the most persistent and strongest vasoconstrictor peptide in the human body and a driver of cell proliferation, which can cause proliferation and stenosis of the vascular wall, leading to hypertrophy and proliferation of vascular smooth muscle.
[0138] Renin: Renin acts on angiotensinogen (ANG) to convert it into inactive angiotensin I (ANG I). Under the action of ACE, angiotensin I is converted into active angiotensin II (ANG II).
[0139] eNOS: The rate-limiting enzyme for NO synthesis, which promotes NO synthesis and helps regulate vasodilation. Therefore, when the concentrations of IL-6, TNF-α, ANP, AII, ALD, ET-1, and Renin in serum decrease, blood pressure tends to decrease, and when the concentration of eNOS increases, blood pressure tends to decrease.
[0140] The test results are shown in Tables 2 and 3.
[0141] Table 2 Contents of substances with blood pressure regulating function in rat serum
[0142]
[0143] Table 3 Continuation of Table 2
[0144]
[0145]
[0146] The results in Table 2 and Table 3 show that in normal blood pressure rats, there are significant differences in the concentrations of IL-6 and TNF-α between the fermentation group and Comparative Example 2; there are significant differences in the concentration of ANP between the fermentation group and Comparative Example 1 and Comparative Example 2. In hypertensive rats, it can be observed that there are significant differences in the concentrations of IL-6, ANP, AII, and ET-1 between the fermentation group and Comparative Example 1 and Comparative Example 2; there is a significant difference in the concentration of TNF-α between the fermentation group and Comparative Example 1; and there is a significant difference in the concentration of ALD between the fermentation group and Comparative Example 2.
[0147] 2.2 Content of antioxidant active substances
[0148] SOD: superoxide dismutase, removes superoxide anion free radicals that are harmful to the body; NO: regulates vasodilation and prevents blood pressure from rising; GSH: glutathione, anti-oxidation, removes free radicals; TAOC: total reactive oxygen species, excessive amounts lead to oxidative stress and cell damage; MDA: malondialdehyde, one of the end products of membrane peroxidation, reflects the rate and intensity of lipid peroxidation in the body. The results are shown in Table 4.
[0149] Table 4 Contents of substances with antioxidant activity in rat serum
[0150]
[0151] The results in Table 4 show that there is no significant difference in the concentration of substances related to antioxidant function in normal blood pressure rats. In hypertensive rats, it can be observed that the SOD and TAOC enzyme activities are significantly different between the fermentation group and comparative example 1 and comparative example 2. There is a significant difference in NO concentration between the fermentation group and comparative example 2.
[0152] 2.3 Differences in blood pressure of rats at different times
[0153] The blood pressure results of rats at different times are shown in Tables 5 and 6.
[0154] Table 5 Blood pressure monitoring of rats
[0155]
[0156] Note: SBP is systolic blood pressure; DBP is diastolic blood pressure; MBP is mean arterial pressure.
[0157] Table 6 Continuation of Table 5
[0158]
[0159]
[0160] Note: SBP is systolic blood pressure; DBP is diastolic blood pressure; MBP is mean arterial pressure.
[0161] In the experimental group of hypertensive rats, the data showed that the blood pressure of the fermentation group was significantly lower than that of the control group 1 and control group 2 in the second week, and the blood pressure of the fermentation group was still significantly lower than that of the control group 1 in the third week; after the fourth week, there was no significant difference in the blood pressure of the rats in the fermentation group, control group 1, control group 2 and captopril positive control. There was no significant difference among the experimental groups in normal blood pressure rats.
[0162] The above experimental results show that the antihypertensive activity of chrysanthemum in hypertensive rats can be improved through microbial fermentation.
[0163] A chrysanthemum fermented product provided by the present invention is a fermented product obtained by using chrysanthemum flowers as materials, and then drying, crushing, screening and soaking, and fermenting with the probiotics. The present invention studies the blood pressure lowering and antioxidant activities of chrysanthemum flowers during the fermentation process using probiotics, and finds that the blood pressure lowering ability of chrysanthemums is increased by fermentation with probiotics. Therefore, the present invention provides a fermented product with chrysanthemum as a substrate and having both blood pressure lowering and antioxidant effects, and a preparation method thereof. The preparation method provided by the present invention can increase the abundance of effective ingredients in the fermented product with chrysanthemum as a substrate compared to chrysanthemum extract. The chrysanthemum fermented product prepared by the preparation method of the chrysanthemum fermented product provided by the present invention has more types of blood pressure lowering active substances, and has more small molecular substances with blood pressure lowering activity such as polysaccharides, small peptides and other substances, thereby improving the blood pressure lowering effect of chrysanthemum extract.
[0164] It should be noted that when the claims of the present invention involve numerical ranges, it should be understood that the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. In order to avoid redundancy, the present invention describes a preferred embodiment.
[0165] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.
[0166] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.
Claims
1. A method for preparing a chrysanthemum fermented product, characterized in that: The method comprises the following steps: The chrysanthemum flowers are dried, crushed and sieved to obtain chrysanthemum powder; adding a solvent to the chrysanthemum powder for soaking to obtain a mixture of chrysanthemum and soaking solution; After the probiotics are activated, they are inoculated into the mixture of the chrysanthemum and the soaking solution for fermentation and culture; After the fermentation is finished, the fermentation liquid is filtered and the chrysanthemum is removed to obtain the chrysanthemum fermented product; The probiotics include any one or more of lactic acid bacteria and yeast.
2. The method for preparing a fermented chrysanthemum product according to claim 1, characterized in that: The inoculation amount of the probiotics is 0.5%wt to 1.5%wt.
3. The method for preparing a fermented chrysanthemum product according to claim 1, characterized in that: The probiotics include lactic acid bacteria and yeast, and the lactic acid bacteria are Lactobacillus; The inoculation volume ratio of the lactobacillus to the yeast is 1:
1.
4. The method for preparing a fermented chrysanthemum product according to claim 3, characterized in that: The lactobacillus is Lactobacillus plantarum; and the yeast is Saccharomyces cerevisiae.
5. The method for preparing a fermented chrysanthemum product according to claim 4, characterized in that: The activation and inoculation methods of the plant lactobacillus and the saccharomyces cerevisiae are as follows: The plant lactobacillus is cultured in an MRS medium at 37° C. for 12 hours to obtain activated plant lactobacillus; The brewer's yeast is cultured in a YPD medium at 30° C. for 32 hours to obtain activated brewer's yeast; After the activated Lactobacillus plantarum and the activated Saccharomyces cerevisiae grow to the logarithmic growth phase, they are inoculated into the mixture of the chrysanthemum and the soaking solution at an inoculation amount of 0.5%wt to 1.5%wt respectively.
6. The method for preparing a fermented chrysanthemum product according to claim 1, characterized in that: The fermentation culture conditions are: 25°C to 35°C, and fermentation for 15d to 30d.
7. The method for preparing a fermented chrysanthemum product according to claim 1, characterized in that: The liquid ratio of the chrysanthemum powder to the added solvent is 0.2 g / mL to 0.4 g / mL; The solvent is water.
8. The fermented chrysanthemum product prepared by the method for preparing a fermented chrysanthemum product according to claim 1.
9. Use of the chrysanthemum fermented product according to claim 8 in improving the antihypertensive effect of chrysanthemum extract.
10. The use according to claim 9, characterized in that: The chrysanthemum fermented product is used for increasing the abundance of antihypertensive active substances in the chrysanthemum extract.