Industrial hempseed meal fermentation product capable of reducing blood pressure as well as preparation method and application of industrial hempseed meal fermentation product

Abstract

The invention provides an industrial hempseed meal fermentation product for reducing blood pressure as well as a preparation method and application thereof, and belongs to the technical field of deep processing of industrial hempseed meal. The method for preparing the industrial hempseed meal fermentation product for reducing blood pressure comprises the following steps: carrying out alkali dissolution and acid precipitation on industrial hempseed meal to obtain industrial hempseed protein, and mixing the industrial hempseed protein and water according to a solid-liquid ratio of 1g: (20-30) mL to obtain a fermentation culture medium; and inoculating a lactobacillus plantarum seed solution into the fermentation culture medium, fermenting at 35-40 DEG C for 65-72 hours to obtain a fermentation solution, centrifuging, and taking a supernatant to obtain the industrial hempseed meal fermentation product. The industrial hempseed meal leavening prepared by the method disclosed by the invention has an obvious function of inhibiting the activity of angiotensin converting enzyme in vitro, shows higher antihypertensive activity in a hypertensive animal in-vivo experiment, and can influence the concentration of a plurality of substances related to the antihypertensive activity.

Description

Technical Field

[0001] The present invention belongs to the technical field of deep processing of industrial hemp seed meal, and particularly relates to a hypotensive industrial hemp seed meal ferment and its preparation method and application. Background Art

[0002] Hypertension is one of the common cardiovascular diseases, which is a phenomenon that the pressure value of blood flowing on the blood vessel wall continuously exceeds the normal value. The diagnostic criteria for adult hypertension are systolic blood pressure ≥ 130 mmHg and / or diastolic blood pressure ≥ 80 mmHg. Hypertension will increase the risks of stroke, coronary heart disease, heart failure and renal failure, and pose a greater harm to human health, and will pose a huge threat to the human body and life. At present, antihypertensive drugs are widely used to treat hypertension and other cardiovascular diseases. The five most commonly used types of antihypertensive drugs include calcium channel blockers, diuretics, angiotensin-converting enzyme inhibitors, angiotensin II inhibitors / angiotensin receptor blockers, and β-blockers. Although the commonly used antihypertensive drugs at present can control the symptoms of hypertension to a certain extent, most of them are chemical agents and more or less have some side effects. For example, captopril and lisinopril have the possibility of causing side effects such as cough and angioedema. Therefore, there is an urgent need in this field to provide a safer and milder substitute derived from plants or animals.

[0003] Industrial hemp seed meal is the residue (seed meal) after extracting oil from industrial hemp seeds. In this field, industrial hemp seed meal is mostly used as feed for raising livestock and poultry, and there is still a lack of relevant research on the deep processing of industrial hemp seed meal. Summary of the Invention

[0004] In view of this, the purpose of the present invention is to provide a method for preparing an industrial hemp seed meal ferment with a hypotensive effect.

[0005] In order to achieve the above-mentioned invention purpose, the present invention provides the following technical solutions:

[0006] The present invention provides a method for preparing a hypotensive industrial hemp seed meal ferment, which includes the following steps: subjecting industrial hemp seed meal to alkali dissolution and acid precipitation to obtain industrial hemp protein, mixing the industrial hemp protein and water according to a material-liquid ratio of 1 g:(20 - 30) mL to obtain a fermentation medium; inoculating a Lactobacillus plantarum seed solution into the fermentation medium, fermenting at 35 - 40 °C for 65 - 72 h to obtain a fermentation broth, centrifuging to take the supernatant to obtain an industrial hemp seed meal ferment.

[0007] Preferably, the alkali dissolution and acid precipitation includes the following steps: mixing industrial hemp seed meal with water, adjusting the pH to 10, stirring and extracting, centrifuging to take the supernatant and adjusting the pH to 3, standing, centrifuging, taking the precipitate, washing with water, and freeze-drying to obtain industrial hemp protein.

[0008] Preferably, the conditions for stirring extraction are: extraction at 35-45 °C for 1.5-2.5 h.

[0009] Preferably, the concentration of the Lactobacillus plantarum seed liquid is 1×10 5 ~1×10 7 CFU / mL.

[0010] Preferably, the culture medium of the Lactobacillus plantarum seed liquid is M17 culture medium.

[0011] Preferably, the pH of the fermentation medium is 6.5-7.5.

[0012] Preferably, the volume ratio of the Lactobacillus plantarum seed liquid to the fermentation medium is 4-6 mL:100 mL.

[0013] The present invention also provides an industrial hemp seed meal fermentate prepared by the above method.

[0014] The present invention also provides the application of the above method or the above industrial hemp seed meal fermentate in the preparation of products for lowering blood pressure or helping to maintain a healthy blood pressure level.

[0015] The present invention also provides a blood pressure lowering product, and the raw materials of the product include the above industrial hemp seed meal fermentate.

[0016] Advantages of the present invention:

[0017] The present invention provides a method for preparing an industrial hemp seed meal fermentate. The industrial hemp seed meal fermentate prepared by the method of the present invention has an obvious function of inhibiting the activity of angiotensin converting enzyme (ACE) in vitro. In the in vivo experiment of hypertensive animals, it shows a high blood pressure lowering activity and can affect the concentrations of multiple substances related to blood pressure lowering activity. Description of the drawings

[0018] Figure 1 The ACE inhibition rate of the products fermented by different strains. Among any two groups, if the column labels have the same letter, it indicates that there is no significant difference between the two groups, p≥0.05. If the column labels have different letters, it indicates that there is a significant difference between the two groups, p<0.05. Detailed implementation manners

[0019] The present invention provides a method for preparing a blood pressure lowering industrial hemp seed meal fermentate, including the following steps: dissolving the industrial hemp seed meal with alkali and precipitating with acid to obtain industrial hemp protein, mixing the industrial hemp protein and water according to a material-liquid ratio of 1 g:(20-30) mL to obtain a fermentation medium; inoculating the Lactobacillus plantarum seed liquid into the fermentation medium, fermenting at 35-40 °C for 65-72 h to obtain a fermentation broth, centrifuging to take the supernatant to obtain the industrial hemp seed meal fermentate.

[0020] The present invention has no special limitation on the specific source of industrial hemp seed meal. In the present invention, the alkali dissolution and acid precipitation preferably include the following steps: mixing industrial hemp seed meal with water, adjusting the pH to 10, stirring and extracting, centrifuging to obtain the supernatant, adjusting the pH of the supernatant to 3, standing, centrifuging, washing the precipitate with water, and freeze-drying to obtain industrial hemp protein. In the present invention, the mass-volume ratio of industrial hemp seed meal to water is preferably 1:50, and the water is preferably distilled water. The present invention has no special limitation on the specific reagent for adjusting the pH, and conventional reagents for adjusting the pH in the art can be used. In the present invention, the conditions for the stirring extraction are preferably: extracting at 35-45 °C for 1.5-2.5 h, more preferably extracting at 40 °C for 2 h. After the stirring extraction, it is preferably centrifuged at 1000 r / min for 20 min, and the supernatant is taken and the pH is adjusted to 3. In the present invention, the standing time is preferably 1 h. After standing, it is preferably centrifuged at 4000 r / min for 15 min, and the supernatant is discarded. In the present invention, after washing the precipitate with water, water is preferably added to the precipitate to make up a certain volume, and the pH is adjusted to neutral, and then it is freeze-dried. The present invention has no special limitation on the specific conditions for freeze-drying.

[0021] In the present invention, the material-liquid ratio of industrial hemp protein to water is preferably 1 g:(22-28) mL, more preferably 1 g:(25-26) mL. In the present invention, the pH of the fermentation medium is preferably 6.5-7.5, more preferably 6.8-7.2. In the present invention, the preparation method of the Lactobacillus plantarum seed liquid is preferably: inoculating the activated Lactobacillus plantarum into the seed medium and culturing at 37 °C to obtain the seed liquid; then centrifuging the seed liquid to obtain the precipitate, and adding PBS buffer solution to the precipitate to obtain the Lactobacillus plantarum seed liquid. The concentration of the Lactobacillus plantarum seed liquid is preferably 1×10 5 ~1×10 7 CFU / mL, more preferably 1×10 6 CFU / mL. In the present invention, the seed medium is preferably M17 medium.

[0022] In the present invention, the volume ratio of the Lactobacillus plantarum seed liquid to the fermentation medium is preferably 4-6 mL:100 mL, more preferably 5 mL:100 mL. In the present invention, the fermentation temperature is preferably 36-38 °C, more preferably 37 °C, and the fermentation time is preferably 67-70 h, more preferably 69 h. After the fermentation to obtain the fermentation broth, the supernatant is obtained by centrifugation to obtain the industrial hemp seed meal fermentate. The temperature and rotation speed of the centrifugation are preferably 4 °C and 4000 rpm / min. In the present invention, after obtaining the industrial hemp seed meal fermentate, it is preferably possible to use freeze-drying to make the industrial hemp seed meal fermentate into freeze-dried powder of industrial hemp seed meal fermentate, and the temperature of the freeze-drying is preferably -20 °C.

[0023] The present invention also provides an industrial hemp seed meal fermentate, which is prepared by the above method.

[0024] The present invention also provides the application of the above method or the above industrial hemp seed meal fermentate in the preparation of products for reducing blood pressure or helping to maintain a healthy blood pressure level. In the present invention, the products preferably include drugs and health products. When the product is a drug, the industrial hemp seed meal fermentate plays the role of reducing blood pressure. When the product is a health product, the industrial hemp seed meal fermentate plays the role of helping to maintain a healthy blood pressure level.

[0025] The present invention also provides a blood pressure reducing product, and the raw materials of the product include the above-mentioned industrial hemp seed meal fermentate.

[0026] The technical solutions provided by the present invention will be described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0027] In the following embodiments, unless otherwise specified, they are all conventional methods.

[0028] In the following embodiments, the materials, reagents, etc. used, unless otherwise specified, can all be obtained from commercial channels.

[0029] The sources of some raw materials in the following embodiments are as follows: The industrial hemp meal is provided by Guangxi Oil Factory;

[0030] Protein extraction and fermentation reagents: Sodium hydroxide, hydrochloric acid (analytical pure), M17 medium, and agar are purchased from Sangon Biotech (Shanghai).

[0031] Reagent samples required for in vitro ACE activity detection: Sodium tetraborate, sodium chloride, boric acid, hippuryl-histidyl-leucine, ACE enzyme, sodium hydroxide, hydrochloric acid, ethyl acetate, quinoline, benzenesulfonyl chloride, and absolute ethanol are all purchased from Sangon Biotech Co., Ltd.

[0032] Detection of blood pressure reducing related substances in vivo: The kit developed by Beijing Huaying Biotechnology Research Institute.

[0033] Example 1

[0034] A blood pressure reducing industrial hemp seed meal fermentate is prepared by the following steps:

[0035] (1) Preparation of industrial hemp protein by alkali solution-acid precipitation method: Accurately weigh 50 g of industrial hemp seed meal, add distilled water and mix according to the material-liquid ratio of 1 g:50 mL, adjust the pH to 10, stir and extract at 40 °C for 2 h, then centrifuge at 1000 r / min for 20 min. Take the supernatant and adjust the pH to 3, let it stand for 1 h, then centrifuge at 4000 r / min for 15 min. Discard the supernatant, wash the precipitate, then add water and adjust the pH to neutral, and freeze-dry to obtain industrial hemp protein.

[0036] (2) Mix 5 g of industrial hemp protein with pure water according to the material-liquid ratio of 1 g:20 mL, adjust the pH to 6.5, and sterilize at 121 °C for 20 min to obtain the fermentation medium.

[0037] (3) Inoculate Lactobacillus plantarum (CICC 20982) into the slant medium (the slant medium consists of M17 medium and 1.5% agar), and culture at 37 °C for 16 h for strain activation. Inoculate the activated Lactobacillus plantarum into the seed medium (the seed medium is M17 medium, pH 7.0), and culture at 37 °C for 16 h in a constant temperature incubator to obtain the seed liquid. Centrifuge the seed liquid at 5000 r / min for 10 min, take the precipitate and add PBS buffer to make the concentration of Lactobacillus plantarum seed liquid 1×10 6 CFU / mL.

[0038] (4) Mix the Lactobacillus plantarum seed liquid and the fermentation medium according to the volume ratio of 4 mL:100 mL, stir evenly, ferment at 35 °C for 72 h to obtain the fermentation broth, then centrifuge at 4 °C and 4000 rpm / min for 10 min to take the supernatant to obtain the fermented product of industrial hemp seed meal.

[0039] Example 2

[0040] A hypotensive fermented product of industrial hemp seed meal is prepared by the following steps:

[0041] (1) The same as step (1) of Example 1.

[0042] (2) Mix 5 g of industrial hemp protein with pure water according to the material-liquid ratio of 1 g:30 mL, adjust the pH to 7.5, and sterilize at 121 °C for 20 min to obtain the fermentation medium.

[0043] (3) Inoculate Lactobacillus plantarum (CICC 20982) into the slant medium (the slant medium consists of M17 medium and 1.5% agar), and culture it at 37 °C for 16 h for strain activation. Inoculate the activated Lactobacillus plantarum into the seed medium (the seed medium is M17 medium with a pH of 7.0), culture it in a constant temperature incubator at 37 °C for 16 h to obtain a seed solution. Centrifuge the seed solution at 5000 r / min for 10 min, take the precipitate, and add PBS buffer to make the concentration of the Lactobacillus plantarum seed solution 1×10 5 CFU / mL.

[0044] (4) Mix the Lactobacillus plantarum seed solution and the fermentation medium evenly at a volume ratio of 6 mL:100 mL, ferment at 40 °C for 65 h to obtain a fermentation broth, then centrifuge at 4 °C and 4000 rpm / min for 10 min, and take the supernatant to obtain the fermented product of industrial hemp seed meal.

[0045] Example 3

[0046] A hypotensive fermented product of industrial hemp seed meal is prepared by the following steps:

[0047] (1) The same as step (1) of Example 1.

[0048] (2) Mix 5 g of industrial hemp protein with pure water at a solid-liquid ratio of 1 g:25 mL, adjust the pH to 7.0, sterilize at 121 °C for 20 min to obtain the fermentation medium.

[0049] (3) Inoculate Lactobacillus plantarum (CICC 20982) into the slant medium (the slant medium consists of M17 medium and 1.5% agar), and culture it at 37 °C for 16 h for strain activation. Inoculate the activated Lactobacillus plantarum into the seed medium (the seed medium is M17 medium with a pH of 7.0), culture it in a constant temperature incubator at 37 °C for 16 h to obtain a seed solution. Centrifuge the seed solution at 5000 r / min for 10 min, take the precipitate, and add PBS buffer to make the concentration of the Lactobacillus plantarum seed solution 1×10 7 CFU / mL.

[0050] (4) Mix the Lactobacillus plantarum seed solution and the fermentation medium evenly at a volume ratio of 5 mL:100 mL, ferment at 38 °C for 70 h to obtain a fermentation broth, then centrifuge at 4 °C and 4000 rpm / min for 10 min, and take the supernatant to obtain the fermented product of industrial hemp seed meal.

[0051] Example 4

[0052] The difference from Example 1 is that Lactobacillus plantarum in step (3) is replaced with Bacillus subtilis, Bacillus licheniformis, and Bacillus natto, respectively (the activation of different strains and the preparation conditions of the seed solution adopt the conventional methods of each strain). The concentrations of the obtained seed solutions of different strains are all 1×10 6 CFU / mL, and the remaining steps are the same as those in Example 1. Fermented products of industrial hemp seed meal obtained by fermentation with different strains are obtained.

[0053] Taking the unfermented product as the blank control group, that is, steps (1) and (2) are the same as steps (1) and (2) in Example 1, the difference is that steps (3) and (4) are omitted, no strain is added, and after standing for the same time under the same conditions, it is centrifuged at 4℃ and 4000 rpm / min for 10 min to take the supernatant to obtain industrial hemp protein. This blank control group is denoted as the unfermented group.

[0054] Measure the in vitro ACE inhibition rates of the fermented products of industrial hemp seed meal obtained by fermentation with different strains, the fermented product of industrial hemp seed meal obtained in Example 1, the industrial hemp protein obtained in the unfermented group, and captopril. The specific measurement method is as follows:

[0055] Drug preparation: 0.1 mol / L boric acid buffer solution (BBS buffer solution), and its preparation method is as follows: Solution 1: 1.9068 g of sodium tetraborate and 3.5064 g of sodium chloride, add water to make up to 200 mL. Solution 2: 1.2366 g of boric acid and 3.5064 g of sodium chloride, add water to make up to 200 mL. Mix the two solutions of Solution 1 and Solution 2, adjust the pH to 8.3, and store at 4℃.

[0056] The preparation method of 5 mmol / L hippuryl-histidyl-leucine (HHL) solution is: dissolve 25 mg of hippuryl-histidyl-leucine in 11.64 mL of the above BBS buffer solution, and store at -20℃. Aliquot with 2 mL centrifuge tubes, 500 μL per tube.

[0057] The preparation method of 0.1 U N / mL ACE solution is: dissolve 0.1 U N ACE enzyme in 1 mL of BBS buffer solution, and store at -80℃. Aliquot with 0.2 mL centrifuge tubes, 50 μL per tube.

[0058] The preparation method of 1 M HCL is: add 8.5 mL of 36.5% HCL by volume to water to make 100 mL. During the preparation process, add water first and then HCL.

[0059] Experimental steps:

[0060] Treatment of the fermented sample: Freeze-dry 1 L of the prepared industrial hemp seed meal fermented product or industrial hemp protein sample at -20℃ overnight to obtain the freeze-dried powder of the industrial hemp seed meal fermented product or the freeze-dried powder of the industrial hemp protein as the sample to be measured, and captopril is directly used as the sample to be measured.

[0061] Take three groups of 10 mL centrifuge tubes, which are used for the blank group, the sample group and the control group respectively. The corresponding centrifuge tubes are called the blank tube, the sample tube and the control tube respectively. In the blank group, 50 μL of HHL and 20 μL of the sample to be tested (concentration: 25 mg / mL) are mixed; in the sample group, 50 μL of HHL and 20 μL of the sample to be tested (concentration: 25 mg / mL) are mixed; in the control group, 50 μL of HHL and 20 μL of BBS buffer are mixed. Each of the above groups reacts at 37 °C for 5 min. Then, 10 μL of ACE is added to the centrifuge tubes of the sample group and the control group, and 10 μL of ACE inactivated by HCL is added to the blank tube. Mix well and react at 37 °C for 30 min. 100 μL of HCL is added to the centrifuge tubes of the sample group, the control group and the blank group to terminate the reaction. Borate buffer solution is added to make up to 0.5 mL. Under light-shielded conditions, 600 μL of quinoline is added and mixed on a vortex mixer for 10 s. 200 μL of benzenesulfonyl chloride is added to each of the above groups, and the tube plugs are immediately tightened and mixed on a vortex mixer for 20 s. After a 30-min light-shielded water bath at 30 °C, 3.7 mL of absolute ethanol is added respectively, and then placed in the dark at 30 °C for 30 min. Take 0.2 mL of the reaction mixture and add it to a 96-well flat-bottom culture plate, and measure the absorbance at a wavelength of 492 nm with an enzyme-linked immunosorbent assay (ELISA) reader. Calculate the ACE inhibition rate using the following formula:

[0062]

[0063] In the formula: A a —Absorbance in the presence of both ACE and the sample (sample); A b —Absorbance without the sample participating in the reaction (control); A c —Absorbance without ACE enzyme participating in the reaction (blank). It should be noted that each sample corresponds to a control and a blank. Here, "each sample" refers to the freeze-dried powder of industrial hemp protein or the freeze-dried powder of the fermented product of industrial hemp seed meal. Each of the above experiments is carried out in 3 parallel experiments, and the results are expressed as the mean ± standard deviation.

[0064] The results are as Figure 1 shown. The products obtained by fermentation with different strains have different blood pressure-lowering activities. The fermentation product of Lactobacillus plantarum of the present invention has a relatively high ACE inhibition rate, indicating that the fermented product of industrial hemp seed meal obtained by the preparation method of the present invention has a relatively high blood pressure-lowering activity. Although Figure 1 the blood pressure-lowering activity of the product obtained by fermentation with Bacillus subtilis in

[0065] Example 5

[0066] 5.1 The fermented product of industrial hemp seed meal obtained in Example 3 and the unfermented group were respectively used for the following animal experiments. The difference between the unfermented group and Example 3 was that steps (3) and (4) were omitted, no strains were added, and after standing under the same conditions for the same time, the supernatant was taken by centrifuging at 4°C and 4000 rpm / min for 10 min to obtain industrial hemp protein, and the rest was the same as in Example 3.

[0067] 5.2 The experimental animals were SPF-grade Wistar rats, male, with a body weight of 0.20 ± 0.02 kg, provided by Changchun Yisi Experimental Animal Technology Co., Ltd.

[0068] 5.3 Drugs and reagents: L-NAME was purchased from Shanghai Yuanye Bio-Technology Co., Ltd. Captopril was purchased from Shanghai Xudong Haipu Pharmaceutical Co., Ltd.

[0069] 5.4 Experimental methods:

[0070] 5.4.1 Preparation of the test drugs:

[0071] 1 L of the industrial hemp seed meal fermented product obtained in Example 3 or the industrial hemp protein sample prepared from the unfermented group was freeze-dried overnight at -20°C to obtain the freeze-dried powder of the industrial hemp seed meal fermented product or the freeze-dried powder of industrial hemp protein, and they were respectively formulated into solutions with a concentration of 25 mg / mL for use.

[0072] L-NAME solution: It was formulated into a medicinal solution with a content of 1.8 mg / mL by adding distilled water for use.

[0073] Captopril solution: Captopril tablets were ground into fine powder with a mortar and formulated into a medicinal solution with a content of 0.5 mg / mL by adding distilled water for use.

[0074] 5.4.2 Animal grouping and drug administration:

[0075] Wistar rats were allowed to drink water and eat freely and were adaptively fed for one week.

[0076] According to the principle of random grouping, the Wistar rats after one week of adaptive feeding were divided into a fermentation group, an unfermented group, and a captopril group, with a drug administration dose of 250 mg / kg, and 9 rats in each group.

[0077] 5.4.3 Experiment on the effect of the industrial hemp seed meal fermented product obtained in Example 3 on the blood pressure of hypertensive rats

[0078] 5.4.3.1 Establishment of a hypertensive rat model

[0079] Healthy male Wistar rats with a body weight in the range of 200 ± 20 g were provided with regular feed and distilled water and adaptively fed for one week. After the adaptive feeding ended, at a dose of 18 mg / kg, every morning at 9 o'clock, the drug L-NAME was given by gavage to establish the model, and the drug induction continued for 4 weeks. The blood pressure of the rats was measured once a week. After the drug induction ended, blood pressure was measured using a non-invasive blood pressure monitor. If the systolic blood pressure of the rats was ≥ 140 mmHg, it indicated that the model was successfully established. The difference between the normal blood pressure rats and the model rats was that instead of giving the drug L-NAME, the same dose of water was given by gavage.

[0080] 5.4.3.2 Animal grouping and drug administration

[0081] Wistar rats with successfully established hypertension models were selected and grouped according to the random principle. The positive control group of captopril was at a dose of 1 mg / kg, and the dosing doses of the fermented product group and the non-fermented control group of hypertensive rats were 250 mg / kg, with 9 rats in each group. The normal blood pressure rats were randomly divided into the fermented product group and the non-fermented control group, with a dosing dose of 250 mg / kg and 9 rats in each group.

[0082] 5.4.3.3 Measurement of rat tail artery pressure and weekly body weight

[0083] Every week at a fixed time, the rat tail artery pressure was measured using a BP-100A non-invasive blood pressure measurement and analysis system while the rats were awake and quiet. The systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MBP) of the rats in the recording system were measured three times continuously, and the average value was taken.

[0084] 5.4.3.4 Collection of specimens

[0085] After the rats were given drugs by gavage, they were anesthetized by intraperitoneal injection of 3 mL / 1 kg of 10% chloral hydrate solution by volume percentage. 5 mL of blood was quickly taken from the rat heart and placed in a blood collection tube without anticoagulant. It was centrifuged at 3500 r / min at 4 °C for 10 min, and the supernatant was stored at -20 °C for measuring the physiological and biochemical indexes in the rat serum. After taking the blood, the rats were sacrificed. The contents of angiotensin (AII), renin, aldosterone (ALD), endothelial nitric oxide synthase (eNOS), tumor necrosis factor α (TNFα), interleukin-6 (IL-6), endothelin (ET-1), and atrial natriuretic peptide (ANP) in the rat serum were all measured by Beijing Huaying Biotechnology Research Institute using the kits developed by Beijing Huaying Biotechnology Research Institute.

[0086] 5.4.3.5 Experimental results

[0087] The results of in vivo experiments are shown in Table 1 and Table 2. As can be seen from Table 1 and Table 2, in normal blood pressure rats, there were no significant differences in the concentrations of various substances between the fermented group and the unfermented group. In hypertensive rats, there were significant differences in the concentrations of the 8 substances detected between the fermented group and the unfermented group.

[0088] Table 1 Content of functional substances regulating blood pressure in rat serum

[0089]

[0090]

[0091] Note: In the normal blood pressure group, the fermented group was compared with the unfermented group; in the hypertensive group, the fermented group, the unfermented group and the captopril group were compared. Different superscript letters indicate significant differences.

[0092] Table 2 Content of functional substances regulating blood pressure in rat serum

[0093]

[0094] Note: In the normal blood pressure group, the fermented group was compared with the unfermented group; in the hypertensive group, the fermented group, the unfermented group and the captopril group were compared. Different superscript letters indicate significant differences.

[0095] The blood pressure results of rats at different times are shown in Table 3. It can be seen that in the experimental group of hypertensive rats, the blood pressure of rats in the fermented group was significantly lower than that in the unfermented group at the 2nd week and the 3rd week. There were no significant differences among the experimental groups in normal blood pressure rats.

[0096] Table 3 Blood pressure monitoring results of rats

[0097]

[0098] Note: In the normal blood pressure group, the fermented group was compared with the unfermented group; in the hypertensive group, the fermented group, the unfermented group and the captopril group were compared. Different superscript letters indicate significant differences.

[0099] Comparative Example 1

[0100] The difference from Example 2 is that the industrial hemp seed meal was replaced with rapeseed meal (purchased from Jiunong Technology Co., Ltd., Shijiazhuang, Hebei), and the rest was the same as in Example 2.

[0101] A non-fermented control group was set up, denoted as the unfermented rapeseed meal group: The difference from Example 2 is that the industrial hemp seed meal was replaced with rapeseed meal (purchased from Jiunong Technology Co., Ltd., Shijiazhuang, Hebei) and fermentation was not carried out in step (4), and the rest was the same as in Example 2.

[0102] Comparative Example 2

[0103] The difference from Example 2 is that industrial hemp seed meal is replaced with skim milk (Mengniu), and the fermentation medium is 150 mL of skim milk, and the rest is the same as in Example 2.

[0104] A control group without fermentation was set up, denoted as the unfermented milk group: The difference from Example 2 is that industrial hemp seed meal is replaced with skim milk (Mengniu) and fermentation is not carried out in step (4), and the rest is the same as in Example 2.

[0105] Comparative Example 3

[0106] The difference from Example 2 is that steps (3) and (4) are omitted, no strain is added, and after placing under the same conditions for the same time, the supernatant is taken by centrifuging at 4°C and 4000 rpm / min for 10 min to obtain industrial hemp protein, denoted as the unfermented industrial hemp seed meal group.

[0107] The in vitro ACE inhibitory rates of the products of Example 2 and Comparative Examples 1 to 3 were measured respectively. The specific measurement method was the same as in Example 4, and the results are shown in Table 4. It can be seen that the products prepared from different raw materials have different ACE inhibitory rates, and the ACE inhibitory rate of the raw materials can be improved after fermentation with the Lactobacillus plantarum of the present invention.

[0108] Table 4 In vitro ACE inhibitory rates after fermentation with different raw materials

[0109] Test substance ACE inhibition rate Unfermented group of industrial hemp seed meal 30.63％±2.32％ Fermented group of industrial hemp seed meal 79.44％±3.18％ Unfermented group of rapeseed meal 15.12％±2.44 Fermented group of rapeseed meal 37.87％±5.19％ Unfermented group of milk 0 Fermented group of milk 13.97％±3.73％

[0110] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A method for preparing a fermented product of industrial hemp seed meal with blood pressure lowering effect, characterized in that, It includes the following steps: obtaining industrial hemp protein by alkali dissolution and acid precipitation of industrial hemp seed meal, mixing the industrial hemp protein and water at a solid-liquid ratio of 1 g:(20-30) mL to obtain a fermentation medium; inoculating a Lactobacillus plantarum seed solution into the fermentation medium, fermenting at 35-40 °C for 65-72 h to obtain a fermentation broth, centrifuging to take the supernatant, and obtaining an industrial hemp seed meal fermentate.

2. The method according to claim 1, characterized in that The alkali dissolution and acid precipitation includes the following steps: mixing industrial hemp seed meal with water, adjusting the pH to 10, stirring for extraction, centrifuging to take the supernatant and adjusting the pH to 3, standing, centrifuging, washing the precipitate with water, and freeze-drying to obtain industrial hemp protein.

3. The method according to claim 2, wherein, The conditions for the stirring extraction are: extracting at 35-45 °C for 1.5-2.5 h.

4. The method according to claim 1, characterized in that The concentration of the Lactobacillus plantarum seed liquid is 1×10 5 ~1×10 7 CFU / mL.

5. The method according to claim 1, characterized in that The medium of the Lactobacillus plantarum seed solution is M17 medium.

6. The method according to claim 1, wherein The pH of the fermentation medium is 6.5-7.

5.

7. The method according to claim 1, wherein The volume ratio of the Lactobacillus plantarum seed solution to the fermentation medium is 4-6 mL:100 mL.

8. An industrial hemp seed meal ferment, characterized in that, Prepared by the method according to any one of claims 1-7.

9. Use of the method according to any one of claims 1-7 or the industrial hemp seed meal fermentate according to claim 8 in the preparation of a product for reducing blood pressure or helping to maintain a healthy blood pressure level.

10. A blood pressure lowering product, characterized in that, The raw materials of the product include the industrial hemp seed meal fermentate according to claim 8.

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