Kudzuvine root fermentation product with blood pressure lowering effect and preparation method thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicine fermentation, and particularly relates to a radix puerariae fermented product with a blood pressure reducing effect and a preparation method thereof. The preparation method comprises the following steps: (1) adding water into kudzuvine root powder, uniformly mixing, and carrying out ultrasonic treatment to obtain a mixed solution; (2) sterilizing the mixed solution to obtain a sterilized solution; and (3) adding a leavening agent into the sterilized solution, fermenting at 25-35 DEG C for 24-72 hours to obtain a fermentation solution, and performing solid-liquid separation on the fermentation solution to obtain a fermentation supernatant, namely a fermentation product. According to the method, the content of puerarin, genistein and total triterpenes in the radix puerariae fermentation liquor can be increased, and a very good blood pressure reducing effect is achieved.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine fermentation, and particularly relates to a kudzu root fermentation product with blood pressure lowering effect and a preparation method thereof. Background Art

[0002] Hypertension (also known as high blood pressure) is a phenomenon in which the pressure on the blood vessel wall caused by blood flow is continuously higher than normal. According to the "Chinese Hypertension Clinical Practice Guidelines", the diagnostic criteria for adult hypertension are systolic blood pressure ≥130mmHg and / or diastolic blood pressure ≥80mmHg. The causes of hypertension are mostly genetic factors and unhealthy lifestyles, such as high-salt diet, excessive drinking, long-term mental stress and lack of physical activity. Typical symptoms of hypertension include headache, fatigue or restlessness, arrhythmia, palpitations and tinnitus. However, many hypertensive patients may have other complications without any symptoms, such as stroke, blurred vision, loss of consciousness, amnesia, etc., so hypertension is called the "silent killer."

[0003] Pueraria root is the dried root of Pueraria lobata (Willd.) Ohwi, a plant of the Leguminosae family. It is a commonly used traditional Chinese medicine with the effects of relieving muscle and fever, clearing rashes, promoting body fluid and quenching thirst, and raising yang and stopping diarrhea. Modern studies have shown that Pueraria root has pharmacological effects such as sobering up, protecting the liver, lowering blood sugar, lowering blood pressure, anti-inflammatory, anti-oxidation, and relieving osteoporosis. Pueraria root contains rich nutrients and bioactive ingredients, such as flavonoids, coumarins, triterpenes, triterpenoid saponins, and Pueraria polysaccharides. Among them, flavonoid compounds are mostly isoflavones, including puerarin, daidzein, daidzein, genistein, etc.; triterpenes and triterpenoid saponins, including soyasaponin A, soyasaponin B, and soyasaponin I, etc.

[0004] Modern fermentation technology is widely used in traditional Chinese medicine. After fermentation, traditional Chinese medicine changes its original properties, enhances or produces new effects, and expands the variety of medicines to meet clinical drug needs. During the fermentation process, microorganisms can metabolize and produce different lignin-degrading enzymes and hydrolases to degrade fibers, causing structural damage to the cell wall and releasing active substances from it. Lactic acid bacteria (LAB) are an important part of the human intestinal microbiome, which can regulate the normal gastrointestinal flora of the body and maintain the balance of intestinal microecology. The use of fermentation technology to organically combine the efficacy of lactic acid bacteria and traditional Chinese medicines can increase the dissolution and conversion efficiency of the effective ingredients of traditional Chinese medicines and enhance the function of the medicine. Summary of the invention

[0005] To this end, the present invention provides a kudzu root fermentation product, which uses kudzu root Chinese medicine as a starting material and is fermented by ultrasound-assisted water extraction. The contents of puerarin, genistein and total triterpenes in the kudzu root fermentation product are improved, and the obtained kudzu root fermentation product has a good blood pressure lowering effect.

[0006] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0007] A method for preparing a Pueraria lobata fermented product having blood pressure lowering effect comprises the following steps:

[0008] (1) adding kudzu root powder to water and mixing evenly, and performing ultrasonic treatment to obtain a mixed solution;

[0009] (2) sterilizing the mixed solution to obtain a sterilized solution;

[0010] (3) Adding a fermentation agent to the sterilized liquid, fermenting at 25-35° C. for 24-72 hours to obtain a fermentation liquid, and the fermentation supernatant obtained by solid-liquid separation of the fermentation liquid is the fermentation product.

[0011] The fermentation agents include Bifidobacterium adolescentis and Lactobacillus mucilaginosus fermentation.

[0012] Preferably, the weight ratio of the kudzu root powder to the leavening agent is (500-1000):1.

[0013] Further preferably, the weight ratio of the kudzu root powder to the leavening agent is 800:1.

[0014] More preferably, the weight ratio of the Bifidobacterium adolescentis to the fermented Lactobacillus mucus is (3-7): (1-5).

[0015] More preferably, the weight ratio of Bifidobacterium adolescentis to Lactobacillus mucosa fermentation is 1:1.

[0016] More preferably, the viable bacterial count of the Bifidobacterium adolescentis is 100 billion to 400 billion CFU / g, and the viable bacterial count of the fermented Lactobacillus mucus is 100 billion to 400 billion CFU / g.

[0017] Preferably, the weight ratio of water to kudzu root powder in step (1) is (5-12):1.

[0018] Further preferably, the weight ratio of water to kudzu root powder is 8:1.

[0019] Preferably, the ultrasonic temperature in step (1) is 50-75°C, and the ultrasonic time is 30-60 min.

[0020] Preferably, in step (2), the sterilization temperature is 90-105° C. and the sterilization time is 50-100 min.

[0021] Preferably, the method further comprises the step of crushing the kudzu root into kudzu root powder.

[0022] The Pueraria lobata fermented product obtained according to the above preparation method is used in the preparation of products with blood pressure lowering effect.

[0023] The technical solution of the present invention has the following advantages:

[0024] The present invention provides a pueraria root fermented product with blood pressure lowering effect. The pueraria root is used as a starting material. After ultrasonic-assisted water extraction and fermentation with Bifidobacterium adolescentis and Lactobacillus mucosa, the effective ingredients in the pueraria root can be released. In the fermentation product obtained by the method, the contents of puerarin, genistein and total triterpenes are effectively improved, and the product has a good blood pressure lowering effect.

[0025] 2 The method for preparing the Pueraria lobata fermented product with blood pressure lowering effect provided by the present invention has few process steps and is suitable for industrial production. The fermentation product can be prepared into powder, etc. and added as a raw material to blood pressure lowering products. DETAILED DESCRIPTION

[0026] To make the purpose, technical solutions and advantages of the present invention clearer, the embodiments of the present invention are described in further detail below. Based on the embodiments disclosed in the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present disclosure.

[0027] If no specific experimental steps or conditions are specified in the examples, the experiments can be carried out according to the conventional steps or conditions described in the literature in the field. All reagents without manufacturer specified are conventional reagent products that can be obtained through commercial purchase.

[0028] In the following examples, the source information of some products is as follows:

[0029] Bifidobacterium adolescentis was purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd.; fermented mucinous Lactobacillus, plantarum Lactobacillus and Lactobacillus delbrueckii subsp. bulgaricus were purchased from Jiangsu Xinshenao Biotechnology Co., Ltd., including fermented mucinous Lactobacillus CQPC07, plantarum Lactobacillus CQPC03 and Lactobacillus delbrueckii subsp. bulgaricus.

[0030] Example 1

[0031] This embodiment provides a method for preparing a Pueraria lobata fermented product having the effect of lowering blood pressure, comprising the following steps:

[0032] (1) grinding Pueraria root into Pueraria root powder, and passing through a 200-mesh sieve to obtain Pueraria root powder;

[0033] (2) 500 g of kudzu root powder was added with 4000 mL of water and mixed evenly. After ultrasonic extraction at 60° C. for 45 min, the pH was adjusted to 6.5 with sodium bicarbonate to obtain a mixed solution;

[0034] (3) sterilizing the mixed solution at 98°C for 70 min and then cooling the temperature to 30°C to obtain a sterilized solution;

[0035] (4) Add 0.3125 g of Bifidobacterium adolescentis and 0.3125 g of Lactobacillus mucilaginosus to the sterilized liquid, and ferment at 30° C. for 72 h to obtain a fermentation liquid. The fermentation supernatant obtained by solid-liquid separation of the fermentation liquid is the fermentation product.

[0036] The number of viable bacteria of Bifidobacterium adolescentis is 300 billion CFU / g, and the number of viable bacteria of Lactobacillus mucus fermentation is 300 billion CFU / g.

[0037] Example 2

[0038] This embodiment provides a method for preparing a Pueraria lobata fermented product having the effect of lowering blood pressure, comprising the following steps:

[0039] (1) grinding Pueraria root into Pueraria root powder, and passing through a 200-mesh sieve to obtain Pueraria root powder;

[0040] (2) 440 g of kudzu root powder was added to 4800 mL of water and mixed evenly. After ultrasonic extraction at 75° C. for 30 min, the pH was adjusted to 6 with sodium bicarbonate to obtain a mixed solution;

[0041] (3) sterilizing the mixed solution at 105° C. for 50 min and then cooling the temperature to 35° C. to obtain a sterilized solution;

[0042] (4) Add 0.50 g of Bifidobacterium adolescentis and 0.23 g of Lactobacillus mucilaginosus to the sterilized liquid, and ferment at 35° C. for 24 h to obtain a fermentation liquid. The fermentation supernatant obtained by solid-liquid separation of the fermentation liquid is the fermentation product.

[0043] The number of viable bacteria of Bifidobacterium adolescentis is 300 billion CFU / g, and the number of viable bacteria of Lactobacillus mucus fermentation is 300 billion CFU / g.

[0044] Example 3

[0045] This embodiment provides a method for preparing a Pueraria lobata fermented product having the effect of lowering blood pressure, comprising the following steps:

[0046] (1) grinding Pueraria root into Pueraria root powder, and passing through a 200-mesh sieve to obtain Pueraria root powder;

[0047] (2) 400 g of kudzu root powder was added with 4800 mL of water and mixed evenly. After ultrasonic extraction at 50° C. for 60 min, the pH was adjusted to 7 with sodium bicarbonate to obtain a mixed solution;

[0048] (3) sterilizing the mixed solution at 90°C for 100 min and then cooling the temperature to 25°C to obtain a sterilized solution;

[0049] (4) Add 0.30 g of Bifidobacterium adolescentis and 0.10 g of Lactobacillus mucilaginosus to the sterilized liquid, and ferment at 25° C. for 48 h to obtain a fermentation liquid. The fermentation supernatant obtained by solid-liquid separation of the fermentation liquid is the fermentation product.

[0050] The number of viable bacteria of Bifidobacterium adolescentis is 300 billion CFU / g, and the number of viable bacteria of Lactobacillus mucus fermentation is 300 billion CFU / g.

[0051] Example 4

[0052] This embodiment provides a method for preparing a Pueraria lobata fermented product having the effect of lowering blood pressure, comprising the following steps:

[0053] (1) grinding Pueraria root into Pueraria root powder, and passing through a 200-mesh sieve to obtain Pueraria root powder;

[0054] (2) 600 g of kudzu root powder was added with 3000 mL of water and mixed evenly. After ultrasonic extraction at 65° C. for 50 min, the pH was adjusted to 6.8 with sodium bicarbonate to obtain a mixed solution;

[0055] (3) sterilizing the mixed solution at 95° C. for 80 min and then cooling the temperature to 32° C. to obtain a sterilized solution;

[0056] (4) Add 0.70 g of Bifidobacterium adolescentis and 0.50 g of Lactobacillus mucilaginosus to the sterilized liquid, and ferment at 32° C. for 60 h to obtain a fermentation liquid. The fermentation supernatant obtained by solid-liquid separation of the fermentation liquid is the fermentation product.

[0057] The number of viable bacteria of Bifidobacterium adolescentis is 300 billion CFU / g, and the number of viable bacteria of Lactobacillus mucus fermentation is 300 billion CFU / g.

[0058] Comparative Example 1

[0059] This comparative example provides a method for preparing a Pueraria lobata fermented product having the effect of lowering blood pressure, comprising the following steps:

[0060] (1) grinding Pueraria root into Pueraria root powder, and passing through a 200-mesh sieve to obtain Pueraria root powder;

[0061] (2) 500 g of kudzu root powder was added to 4000 mL of water and mixed evenly, and the pH was adjusted to 6.5 with sodium bicarbonate to obtain a mixed solution;

[0062] (3) sterilizing the mixed solution at 98°C for 70 min and then cooling the temperature to 30°C to obtain a sterilized solution;

[0063] (4) Add 0.3125 g of Bifidobacterium adolescentis and 0.3125 g of Lactobacillus mucilaginosus to the sterilized liquid, and ferment at 30° C. for 72 h to obtain a fermentation liquid. The fermentation supernatant obtained by solid-liquid separation of the fermentation liquid is the fermentation product.

[0064] The number of viable bacteria of Bifidobacterium adolescentis is 300 billion CFU / g, and the number of viable bacteria of Lactobacillus mucus fermentation is 300 billion CFU / g.

[0065] The difference between this comparative example and Example 1 is that no ultrasonic treatment is performed.

[0066] Comparative Example 2

[0067] The difference between this comparative example and Example 1 is only the type of starter. In this comparative example, the starter consists of 0.625 g of fermented Lactobacillus mucilaginosus, and the rest is the same as Example 1.

[0068] Comparative Example 3

[0069] The difference between this comparative example and Example 1 is only in the type of starter, in which the starter is composed of 0.625 g of Bifidobacterium adolescentis. The rest is the same as Example 1.

[0070] Comparative Example 4

[0071] The difference between this comparative example and Example 1 is that the type of fermentation agent is different, and a fermentation strain Lactobacillus bulgaricus is added. In this comparative example, the fermentation agent is composed of 0.208g Bifidobacterium adolescentis, 0.208g fermentation Lactobacillus mucosa and 0.209g Lactobacillus delbrueckii subsp. bulgaricus. The viable count of Lactobacillus delbrueckii subsp. bulgaricus is 300 billion CFU / g, and the rest is the same as Example 1.

[0072] Comparative Example 5

[0073] The difference between this comparative example and Example 1 is that the type of fermentation agent is different, and Lactobacillus plantarum is used to replace Lactobacillus mucilaginosus fermentation. In this comparative example, the fermentation agent is composed of 0.3125g Bifidobacterium adolescentis and 0.3125g Lactobacillus plantarum. The viable count of Lactobacillus plantarum is 300 billion CFU / g, and the rest is the same as Example 1.

[0074] Comparative Example 6

[0075] The present comparative example provides a method for preparing a Pueraria root fermented product having the effect of lowering blood pressure, and the preparation method is as follows: (1) grinding Pueraria root into Pueraria root powder, and passing through a 200-mesh sieve to obtain Pueraria root powder; (2) taking 500 g of Pueraria root powder and adding 4000 mL of water to mix evenly, stirring and extracting at 30° C. for 72 h to obtain a mixed solution; (3) separating the mixed solution into a solid-liquid separation to obtain a supernatant, which is the product.

[0076] The difference between this comparative example and Example 1 is that no ultrasonic and fermentation treatments were performed.

[0077] Experimental Example 1

[0078] This experiment was used to determine the contents of puerarin, daidzein, genistein and total triterpenes in Pueraria lobata fermentation product.

[0079] 1.1 Determination of puerarin, daidzein and genistein by HPLC

[0080] Preparation of standard solution: Accurately weigh 10 mg of puerarin, daidzein, and genistein standards, add them to a 25 mL volumetric flask, dissolve them with methanol and dilute to the mark to prepare three standard stock solutions. Accurately measure 0.10, 0.20, 0.40, 1.00, 2.00, 3.00, and 4.00 mL of each standard stock solution, add them to seven 10 mL volumetric flasks, dilute to the mark with 30% ethanol, and filter through a 0.22 μm organic membrane for later use.

[0081] Sample processing:

[0082] The supernatants of Example 1-4 and Comparative Example 1-6 were concentrated and spray-dried into powders. 0.5 g of sample was weighed, 25 mL of 30% ethanol was added, and the sample was weighed. After ultrasonic extraction for 1 h, the sample was weighed again, and the weight difference was supplemented with 30% ethanol. The sample was filtered, and the filtrate was filtered through a 0.22 μm organic membrane for later use.

[0083] Puerarin, daidzein and genistein were purchased from Shanghai Yuanye Biotechnology Co., Ltd. with the product numbers of puerarin B20446 (analytical standard, HPLC≥98%), daidzein B20227 (analytical standard, HPLC≥98%) and genistein B21039 (analytical standard, HPLC≥98%), respectively.

[0084] Chromatographic analysis conditions:

[0085] Detection instrument: Aglient HP 1100 high performance liquid chromatograph coupled with DAD detector (Agilent Technologies, USA); chromatographic column is Hypersil BDS C18 column (250mm×4.6mm, 5μm); mobile phase is methanol-water; gradient elution: 0-10min, 20% methanol; 10-15min, 25% methanol; 15-18min, 40% methanol; 18-22min, 46% methanol; 22-25min, 55 methanol; 25-30min, 65% methanol; 30-35min, 72% methanol; 35-40min, 20% methanol; flow rate is 1.0mL / min; detection wavelength is 250nm; column temperature is 35℃; injection volume is 10μL.

[0086] The standard curve was drawn with the solution mass concentration X (μg / mL) as the abscissa and the peak area Y as the ordinate to obtain the regression equation. The standard curve regression equations of the three standards are shown in Table 1.

[0087] Table 1 Linear regression equations of three isoflavones

[0088] Standard product name Regression equation <![CDATA[R 2 ]]> Linear range (μg / mL) Puerarin Y=13138X+37591 0.9992 0.55～85.45 Daidzein Y=28875X-17334 0.9993 0.29～15.65 Genistein Y=21508X-18868 0.9991 0.24～17.58

[0089] 1.2 Determination of total triterpenoids by perchloric acid-vanillin colorimetric method

[0090] Preparation of standard solution: Take oleanolic acid as the standard, accurately weigh 20 mg of oleanolic acid into a 25 mL volumetric flask, and add methanol solution to dissolve and make up to volume. Accurately pipette 0.5, 1.0, 1.5, 2.0, and 2.5 mL of the reference solution into 25 mL volumetric flasks, accurately add 0.5 mL of 5% vanillin-glacial acetic acid, then add 1.4 mL of perchloric acid, place in a 60°C water bath for 25 min, cool to room temperature with running water, then add 5.0 mL of glacial acetic acid, and add methanol to make up to 25 mL. After standing for 40 min, measure the absorbance at 550 nm.

[0091] The standard curve was drawn with the concentration of the reference solution (g / L) as the abscissa and the absorbance value (A) as the ordinate. The equation of the standard curve was: Y = 12.4352X - 0.0413, R 2 =0.999.

[0092] Oleanolic acid was purchased from Shanghai Yuanye Biotechnology Co., Ltd., product number B20954 (analytical standard, HPLC ≥ 98%)

[0093] Sample processing:

[0094] The supernatants of Example 1-4 and Comparative Example 1-6 were concentrated and spray-dried into powders. 100 mg of the powder was dissolved in 8 mL of isopropanol and extracted for 24 h before centrifugation to obtain the supernatant, which was then fixed to 10 mL for later use.

[0095] 1.3 Analysis of the contents of three isoflavones and total triterpenes in Pueraria lobata fermentation products

[0096] The contents of three isoflavones and total triterpenes in the kudzu root fermentation products of Example 1-4 and Comparative Example 1-6 are shown in Table 2. The experimental data were statistically analyzed using Excel and SPSS27 software, and the results were recorded as "mean ± standard deviation". Variance analysis was used for statistical analysis, and P < 0.05 was considered to be significantly different.

[0097] Table 2 Contents of puerarin, daidzein, genistein and total triterpenes in Pueraria lobata fermentation products (n=3)

[0098]

[0099] Note: Different lowercase letters in the same column indicate significant differences (P<0.05).

[0100] Result analysis: As shown in the results in Table 2, the contents of puerarin, genistein and total triterpenes in Example 1-4 groups were significantly higher than those in Comparative Example 1-6 groups, indicating that ultrasonic treatment and composite strain fermentation can increase the contents of puerarin, genistein and total triterpenes in Pueraria fermentation broth.

[0101] The difference between Comparative Example 1 and Example 1 is that no ultrasonic pretreatment was performed. As shown in Table 2, the contents of each substance in Comparative Example 1 are lower than those in Example 1, and the differences are significant, indicating that post-ultrasonic fermentation is more conducive to the dissolution of puerarin, daidzein, genistein and total triterpenes.

[0102] The difference between Comparative Example 6 and Example 1 is that no ultrasonic fermentation treatment was performed, only a simple water extraction was performed. From the results in Table 2, it can be seen that the content of puerarin, daidzein, genistein, and total triterpenes in Example 1 is significantly higher than that in Comparative Example 6, indicating that ultrasonic pretreatment combined with fermentation by a starter can greatly increase the content of puerarin, daidzein, genistein, and total triterpenes, thereby improving the medicinal value of Pueraria lobata fermentation product.

[0103] The difference between Comparative Examples 2 and 3 and Example 1 is that only a single strain is included. From the results in Table 2, it can be seen that the contents in Comparative Examples 2 and 3 are lower than those in Example 1, and there are significant differences, indicating that the fermentation of the composite strain is more conducive to the dissolution of puerarin, daidzein, genistein and total triterpenes.

[0104] The difference between Comparative Example 4 and Example 1 is that Lactobacillus bulgaricus is added. As shown in Table 2, the puerarin content in Comparative Example 4 is significantly lower than that in Example 1, and the daidzein content is increased compared with that in Example 1, indicating that the addition of the new strain promotes the further decomposition and conversion of puerarin into daidzein.

[0105] The difference between Comparative Example 5 and Example 1 is that fermented mucus Lactobacillus is replaced by plant lactobacillus. From the results in Table 2, it can be seen that the puerarin content in Comparative Example 5 is reduced compared with that in Example 1, and the daidzein content is increased compared with that in Example 1, indicating that the combination of plant lactobacillus and adolescent bifidobacterium can promote the further decomposition of puerarin.

[0106] Shi Chenxu et al. Research progress on chemical components and pharmacological effects of Pueraria lobata [J]. Modern Chinese Medicine. 2021. (23) 12: 2177-2195 disclosed that puerarin, as the main component of Pueraria lobata isoflavones, has received widespread attention. WU W, YANG S, LIUP, et al. Systems pharmacology based strategy to investigate pharmacological mechanisms of Radix puerariae for treatment of hypertension [J]. Front Pharmacol, 2020, 11: 345 disclosed that through pharmacological studies, puerarin is one of the main compounds in Pueraria lobata that has a blood pressure lowering effect. In addition, genistein and daidzin have also been shown to be effective in treating hypertension. The puerarin in Pueraria lobata can be converted into daidzein, daidzin can be converted into daidzein, and genistein can be converted into genistein. Based on the antihypertensive efficacy components and the conversion relationship, the applicant used puerarin content and genistein content as the main reference indicators when selecting strains. As can be seen from the results in Table 2, the present invention can effectively ensure that the fermentation product has a high content of puerarin and genistein by first ultrasound and then fermentation by Bifidobacterium adolescentis and fermentation Lactobacillus mucilaginosus. The applicant speculates that it may be due to the difference in strains and the different secondary metabolites produced, which leads to the fact that puerarin is not easily converted, while genistein can be converted into genistein. In addition, the applicant also found that after ultrasound and fermentation, the total triterpenoid content increased, and lupeol and lupenone in the triterpenoid compounds have the ability to inhibit the activity of protein tyrosine phosphatase 1B (PTP1B), which can be used in products for the treatment of diabetes.

[0107] Experimental Example 2: Blood Pressure Lowering Effect

[0108] 2.1 Experimental groups and drug administration method

[0109] 88 male spontaneously hypertensive (SHR) rats were randomly divided into 11 groups, 8 rats in each group, namely, a blank control group, Example 1-4 groups, and Comparative Example 1-6 groups. Except for the blank control group, which was gavaged with normal saline, the other groups were gavaged with 10 mL / kg of the corresponding Pueraria fermentation clear liquid, and each group of SHR rats was gavaged for 4 weeks.

[0110] Confirmation of dosage: The daily oral dosage of rats was calculated using the kilogram body weight conversion coefficient method. According to the calculation that an adult weighing 60 kg consumes 100 mL of oral solution per day, the crossover conversion coefficient is 6.25, and the daily oral dosage of rats should be (100 mL / 60 kg) × 6.25 = 10.42 mL / kg, which is convenient for calculation. In subsequent experiments, each group was gavaged at a dose of 10 mL / kg.

[0111] 2.2 Index detection method

[0112] The systolic and diastolic blood pressures of SHR rats were measured every other week. After the 4-week drug administration experiment, the SHR rats in each group were anesthetized, and the orbits were removed to collect blood. The blood was centrifuged at 4°C and 5000r / min for 10 minutes to obtain SHR rat serum. The levels of renin (Renin), angiotensin Ⅱ (AngⅡ) and nitric oxide (NO) in the serum of SHR rats were measured according to the instructions.

[0113] 2.3 Analysis of blood pressure results

[0114] The diastolic and systolic blood pressure results of SHR rats in each group are shown in Tables 3 and 4. Since there were significant differences in blood pressure among the rats in the fourth week, the data table only shows the blood pressure values ​​before administration and four weeks after administration.

[0115] From the results, there was no significant difference in diastolic and systolic blood pressure between the rats in each group before administration. Four weeks after administration, the systolic and diastolic blood pressure of the blank control group were significantly higher than those of Examples 1-4 and Comparative Examples 1-6, indicating that the pueraria fermented product has the effect of lowering blood pressure; the diastolic and systolic blood pressure of the rats in Example 1 were significantly lower than those in Comparative Examples 1-6, indicating that after the pueraria is treated with ultrasound and fermentation, the blood pressure lowering effect of the pueraria can be enhanced, giving the pueraria more medicinal value.

[0116] Table 3 Effects of Pueraria lobata fermentation on diastolic blood pressure in SHR rats (X±S)

[0117]

[0118]

[0119] Note: * represents P<0.05 compared with the blank control group, ** represents P<0.01 compared with the blank control group; # represents P<0.05 compared with Example 1, ## represents P<0.01 compared with Example 1.

[0120] Table 4 Effect of Pueraria lobata fermentation on systolic blood pressure of SHR rats (X±S)

[0121]

[0122]

[0123] Note: * represents P<0.05 compared with the blank control group, ** represents P<0.01 compared with the blank control group; # represents P<0.05 compared with Example 1, ## represents P<0.01 compared with Example 1.

[0124] 2.4 Analysis of serum Renin, AngⅡ and NO results

[0125] The renin-angiotensin-aldosterone system (RAAS) belongs to the endocrine system and is an important regulator of blood pressure and water-salt metabolism. Renin in RAAS is mainly secreted by the juxtaglomerular cells of the renal juxtaglomerular arterioles, and can catalyze the conversion of angiotensinogen (AGT) from the liver into angiotensin I (Ang I). Ang I is degraded into Ang II (i.e., the RAAS system is activated) by angiotensin converting enzyme (ACE) from pulmonary epithelial cells in the pulmonary circulation. Ang II has a strong vasoconstrictive effect and can also cause vascular endothelial cell dysfunction through various pathways (inflammation, coagulation mechanism, etc.), promote the formation of vascular endothelial cell inflammation, promote the occurrence of atherosclerosis, and thus aggravate the progression of hypertension.

[0126] The concentrations of Renin and AngⅡ in the serum of SHR rats in each group are shown in Table 5. From the results, compared with the blank control group, the concentrations of Renin and AngⅡ in the serum of Examples 1-4 and Comparative Examples 1-6 were significantly lower than those of the blank control group, indicating that the kudzu root fermented product can reduce the concentrations of Renin and AngⅡ in rat serum, inhibit the activation of the RAAS system, and thus reduce the blood pressure of SHR rats. Compared with Example 1, the concentrations of Renin and AngⅡ in the serum of Comparative Examples 1-6 were significantly higher than those in Example 1, and there was no significant difference between Examples 1-4, indicating that the type and quantity of the fermentation agent strains selected in the examples and the ultrasonic treatment are the best choices, which can enhance the blood pressure lowering effect of kudzu root.

[0127] NO is an active substance secreted by endothelial cells that relaxes blood vessels and can effectively regulate vascular tension. The decrease in its content is one of the important factors that cause hypertension. The results of NO concentration in each group are shown in Table 5. Compared with the blank control group, the NO concentrations of Examples 1-4 and Comparative Examples 1-6 are significantly higher than those of the blank control group; compared with Example 1, there is no significant difference between Examples 1-4, while the NO concentration of Comparative Examples 1-6 is significantly lower than that of Example 1, indicating that the ultrasonic treatment of the examples, the types and quantities of the fermentation strains, increase the NO concentration in the serum of rats, and reduce the blood pressure of rats.

[0128] Table 5 Results of serum Renin, AngⅡ and NO after four weeks of continuous intervention (X±S)

[0129]

[0130]

[0131] Note: * represents P<0.05 compared with the blank control group, ** represents P<0.01 compared with the blank control group; # represents P<0.05 compared with Example 1, ## represents P<0.01 compared with Example 1.

[0132] Through ultrasound and fermentation, Pueraria root inhibits the RAAS system by reducing the concentrations of Renin and AngⅡ in rat serum, and on the other hand, increases the concentration of NO in rat serum, thereby improving the antihypertensive effect in SHR rats.

[0133] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A method for preparing a Pueraria lobata fermented product having blood pressure lowering effect, characterized in that: The steps include: (1) adding kudzu root powder to water and mixing evenly, and performing ultrasonic treatment to obtain a mixed solution; (2) sterilizing the mixed solution to obtain a sterilized solution; (3) adding a fermentation agent to the sterilized liquid, fermenting at 25-35° C. for 24-72 hours to obtain a fermentation liquid, and the fermentation supernatant obtained by solid-liquid separation of the fermentation liquid is the fermentation product; The fermentation agents include Bifidobacterium adolescentis and Lactobacillus mucilaginosus fermentation.

2. The preparation method according to claim 1, characterized in that: The weight ratio of the kudzu root powder to the leavening agent is (500-1000):

1.

3. The preparation method according to claim 1, characterized in that: The weight ratio of the Bifidobacterium adolescentis to the Lactobacillus mucosa fermentation is (3-7): (1-5).

4. The preparation method according to claim 3, characterized in that: The weight ratio of the Bifidobacterium adolescentis to the Lactobacillus mucilaginosus fermentation is 1:

1.

5. The preparation method according to claim 1, characterized in that: In the step (1), the weight ratio of water to kudzu root powder is (5-12):

1.

6. The preparation method according to claim 1, characterized in that: In the step (1), the weight ratio of water to kudzu root powder is 8:

1.

7. The preparation method according to claim 1, characterized in that: The ultrasonic temperature in step (1) is 50-75°C, and the ultrasonic time is 30-60 min.

8. The preparation method according to claim 1, characterized in that: In the step (2), the sterilization temperature is 90-105° C. and the sterilization time is 50-100 min.

9. The preparation method according to claim 1, characterized in that: The method also includes the step of crushing the kudzu root into kudzu root powder.

10. Use of a fermentation product obtained by the preparation method according to any one of claims 1 to 9 in preparing a product having blood pressure lowering effect.

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