CN115747092A - Two lactobacillus plantarum SR37-3 and SR61-2 with significant blood pressure lowering function and application thereof - Google Patents

Abstract

The invention discloses two lactobacillus plantarum SR37-3 and SR61-2 with obvious blood pressure reducing function and application thereof. Lactobacillus plantarum (Lactobacillus plantarum) SR37-3 with the deposit number: GDMCC No:62390. lactobacillus plantarum (Lactobacillus plantarum) SR61-2 with the deposit number: GDMCC No:62389. the two Lactobacillus plantarum strains with the function of reducing blood pressure are obtained by screening, and are named as Lactobacillus plantarum SR37-3 and Lactobacillus plantarum SR61-2. In vitro tests prove that the two lactobacillus plantarum strains have strong angiotensin converting enzyme inhibition rates and no hemolysis and drug resistance; in vivo experiments prove that the two strains have good blood pressure lowering effect, can relieve kidney injury caused by hypertension, can improve cardiovascular health and the like.

Description

Two lactobacillus plantarum SR37-3 and SR61-2 with significant blood pressure lowering function and application thereof

Technical Field

The invention belongs to the technical field of microorganisms, and particularly relates to two lactobacillus plantarum SR37-3 and SR61-2 with obvious blood pressure reducing functions and application thereof.

Background

Cardiovascular disease has been a major public health problem in today's society, and hypertension is one of the major health risk factors leading to cardiovascular disease. The data show that the prevalence rate of adult hypertension in China reaches 27.9 percent, namely, 1 hypertension patient exists in every 4 adults, and the health of people is seriously threatened.

Patients with hypertension have long often required long-term dependence on drugs to control disease progression. The currently commonly used blood pressure lowering drugs include diuretics (thiazines), angiotensin Converting Enzyme Inhibitors (ACEIs), beta-blockers, calcium channel blockers, and Angiotensin II receptor blockers. Among the currently used drugs, the ACEI drugs represented by captopril and enalapril have better clinical curative effects. The ACEI medicine can inhibit ACE activity to reduce angiotensin II production, and inhibit bradykinin degradation for dilating blood vessel to achieve effects of dilating blood vessel and lowering blood pressure. The ACEI medicine has good antihypertensive effect and fewer side effects, and clinical tests prove that the medicine has good safety for patients with hypertension complicated with coronary heart disease, heart failure, cardiac hypertrophy, cardiomyopathy, diabetes, mild proteinuria, mild renal insufficiency and the like. However, the manufacturing of the ACEI drugs at present mainly depends on chemical synthesis, while the synthesized drugs have some adverse reactions, mainly including slight cough, hypotension, renal function reduction, hyperkalemia, angioneurotic edema and the like, and the application of the ACEI drugs is greatly limited due to the toxic and side effects.

Therefore, it is a research focus of people to develop new therapies and find safer and less-side-effect blood pressure lowering products. In recent years, sequencing technology has been revolutionized, and microbial community has been vigorously developed. This hot tide of the study has led us to re-evaluate the human gut for trillions of microorganisms. The intestinal microbiota has many protective and metabolic functions for the health of the host, including the function of food fermentation degradation, the function of digesting polysaccharides that are difficult to digest by the host, the function of synthesizing vitamins, and the like. Especially, intensive research on probiotics has led to the understanding that people no longer only have knowledge on fermented foods such as pickles and yoghourt. The probiotics in the fields of health-care food, special food and the like are well developed, and products such as probiotic granules, capsules and the like appear in succession. Research shows that some probiotics can produce bioactive components with preventing and regulating effect on cardiovascular diseases after fermenting food. The bioactive factors not only have higher safety and smaller side effect, but also can promote the proliferation of beneficial microorganisms in the intestinal tract, regulate the intestinal microecology and further continuously and deeply influence the health of a host.

The application of blood pressure lowering probiotics in food is mainly focused on dairy products. It is well recognized that probiotics such as lactic acid bacteria can hydrolyze proteins in dairy products to produce short peptides, which have the effect of lowering blood pressure. However, the research on the antihypertensive fermented milk at home and abroad is not much. In 2002, seppo et al studied fermented milk produced by Evolus for lowering blood pressure. The blood pressure decreased more significantly during the test period in the Evolus group compared to the control group, but the blood pressure almost returned to the level before the test after two weeks of discontinuation of the intake of the Evolus fermented milk. In 2005, jauhiainen et al, in a randomized, double-blind placebo-controlled study, 94 hypertensive patients ingested Lactobacillus helveticus fermented milk (containing high concentrations of VPP/IPP peptide) twice daily for 10 weeks with a decrease in blood pressure compared to the control group (SBP/DBP difference between the control group and the dry group was-4.1 + -0.9/-1.8 + -0.7 mm Hg). The study of Ahren et al in 2015 found that after the lactobacillus fermented milk product was consumed, the blood pressure of the hypertensive patients was significantly reduced without any side effects. At present, the countries such as Finland, japan and the like have already appeared the blood pressure lowering products, but the domestic blank of the blood pressure lowering probiotic products still exists. The screening performance is excellent, the blood pressure reducing effect is obvious, and the probiotic can be used for producing functional foods, preventing and treating hypertension, regulating intestinal microecology and promoting cardiovascular health, and has great significance for implementing 'healthy Chinese' actions, promoting human health and improving human life quality.

Disclosure of Invention

The invention aims to provide two lactobacillus plantarum strains with the functions of reducing blood pressure, improving hypertension complications and promoting cardiovascular health and application thereof.

In order to achieve the purpose, the invention adopts the technical scheme that: the Lactobacillus plantarum has the functions of reducing blood pressure and promoting cardiovascular health and is named as Lactobacillus plantarum SR37-3 and Lactobacillus plantarum SR61-2.

Lactobacillus plantarum SR37-3 was deposited at 18.4.2022 in Guangdong province collection of microorganisms (GDMCC) with the following addresses: building 5 of first furnance, large yard, 100, building 59, guangdong province, guangzhou, china, zip code: 510070, accession number: GDMCC No:62390.

lactobacillus plantarum SR61-2 was deposited at 18.4.2022 in Guangdong province collection of microorganisms (GDMCC) with the following addresses: building 5 of first furnance, large yard, 100, building 59, guangdong province, guangzhou, china, zip code: 510070, accession number: GDMCC No:62389.

the two Lactobacillus plantarum strains with the function of reducing blood pressure are obtained by screening, and are named as Lactobacillus plantarum SR37-3 and Lactobacillus plantarum SR61-2. In vitro tests prove that the two lactobacillus plantarum strains have strong angiotensin converting enzyme inhibition rates and no hemolysis and drug resistance; in vivo experiments prove that the two strains have good blood pressure lowering effect, can relieve kidney injury caused by hypertension, can improve cardiovascular health and the like.

The invention also claims application of the lactobacillus plantarum SR37-3 or lactobacillus plantarum SR61-2 in preparation of products for reducing blood pressure, relieving serum inflammatory substances and improving kidney injury and promoting cardiovascular health.

The invention also provides a product for reducing blood pressure, relieving serum inflammatory substances and improving kidney injury and promoting cardiovascular health, which contains the lactobacillus plantarum SR37-3 or lactobacillus plantarum SR61-2 as an active ingredient.

Preferably, the product may be a pharmaceutical or functional food.

In a preferred embodiment of the present invention, the functional food further comprises an auxiliary material, such as inulin, resistant starch, fructo-oligosaccharide, or galacto-oligosaccharide.

As a preferred embodiment of the present invention, the functional food is a fermented food product.

The invention also provides a culture method of the lactobacillus plantarum SR37-3 or the lactobacillus plantarum SR61-2, and a proper amount of the lactobacillus plantarum SR37-3 or the lactobacillus plantarum SR61-2 is inoculated into a culture medium for culture.

As a preferred embodiment of the present invention, the medium is an MRS medium.

The invention also claims a specific nucleotide sequence of the lactobacillus plantarum SR37-3 or SR61-2, wherein the specific nucleotide sequence of the lactobacillus plantarum SR37-3 is shown as SEQ ID NO.1, and the specific nucleotide sequence of the lactobacillus plantarum SR61-2 is shown as SEQ ID NO. 2.

Furthermore, the invention also claims a corresponding PCR primer group, wherein the primer group is designed according to the nucleotide sequences shown as SEQ ID NO.1 and SEQ ID NO. 2.

A nucleotide sequence is detected by designing corresponding primer sets for PCR based on the corresponding nucleotide sequences, each primer set including a forward primer and a reverse primer.

As a preferred embodiment of the invention, the primer group for detecting the lactobacillus plantarum SR37-3 comprises nucleotide sequences shown as SEQ ID No.3 and 4, and the nucleotide sequence for detecting the lactobacillus plantarum SR61-2 is shown as SEQ ID No.5 and 6.

The invention also provides a detection method for detecting the lactobacillus plantarum SR37-3 or the lactobacillus plantarum SR61-2, which comprises the following steps:

s1: performing PCR amplification using the primer set;

s2: carrying out gel electrophoresis to detect the amplification product;

s3: observing whether the amplification product is in accordance with the expectation.

As a preferred embodiment of the invention, the PCR amplification system in S1 comprises a PCR Mix, a template DNA, a primer set and sterilized double distilled water.

As a preferred embodiment of the invention, the PCR amplification system is 2X San Taq PCR Mix of 10. Mu.L, template DNA of 1.0. Mu.L, primers of 1. Mu.L each, and sterilized double distilled water to make up the volume to 20. Mu.L.

In a preferred embodiment of the present invention, the PCR amplification procedures in S1 are respectively: pre-denaturation at 94 ℃ for 4min; denaturation at 94 ℃ for 30s; annealing at 70 ℃ for 30s; extension at 72 ℃ for 60s; 35 cycles of denaturation, annealing and extension are carried out; finally, extension is carried out for 5min at 72 ℃. Pre-denaturation at 94 ℃ for 4min; denaturation at 94 ℃ for 30s; annealing at 70 ℃ for 30s; extension at 72 ℃ for 25s; carrying out denaturation, annealing and extension for 37 cycles; finally, extension is carried out for 5min at 72 ℃.

The invention has the following beneficial effects:

1. the probiotics in the invention are excellent strains separated from Xinjiang fermented food.

2. Compared with the traditional blood pressure lowering chemical drugs, the invention has the advantages of no toxic or side effect on the ecological environment, no residual risk, safety to human body and economy.

3. The invention has good effects of reducing blood pressure and promoting cardiovascular health, which is embodied as follows:

a. the lactobacillus plantarum fermented skim milk provided by the invention has higher angiotensin converting enzyme inhibition rate.

b. The strain shows better safety in vivo and in vitro safety experiments.

c. Has certain regulating effect on aspects such as hypertension representation, enzymes, hormones and inflammatory factors in serum and the like in vivo tests, and has the effects of protecting renal injury caused by hypertension and improving cardiovascular health.

4. The invention has proved from the multi-angle of genome and phenotype detection, animal experiment, etc. that there is no potential harm to human body.

5. The invention has proved its effectiveness and safety through animal experiment. The strain resource library is enriched, and meanwhile, the method has great application potential and value in the aspect of developing functional foods for preventing and treating hypertension.

6. The material basis and metabolic pathways of the functions are primarily mined through a metabonomics means.

Lactobacillus plantarum SR37-3 was deposited in 18 months 4 of 2022 at GDMCC in the collection of microorganisms and cell cultures of guangdong province (GDMCC) address: building 5 of first furios middle way 100 large yard 59, guangdong province, guangzhou, zip code: 510070, accession number: GDMCC No:62390.

lactobacillus plantarum SR61-2 was deposited at 18.4.2022 in Guangdong province collection of microorganisms (GDMCC) with the following addresses: building 5 of first furnance, large yard, 100, building 59, guangdong province, guangzhou, china, zip code: 510070, accession number: GDMCC No:62389.

drawings

FIG. 1 shows the in vivo blood pressure lowering effect of two strains of Lactobacillus plantarum, and the data are expressed using Mean. + -. SE.

FIG. 2 shows the result of ELISA detection of serum in vivo experiment.

FIG. 3 is a graph of a multivariate statistical analysis of UPLC-MS/MS data, LC-MS analysis of a sample of serum from hypertensive rats (a-f); PCA score distribution for the W, LN + SR37-3 and SR61-2 groups in positive model (a) and negative model (b); alignment test pattern of OPLS-DA model between metabolomic datasets W and LN, positive mode (R2 = (0,0.989); Q2= (0, -0.00648)) (c) and negative mode ((R2 = (0,0.876); Q2= (0, -0.458)) (d); OPLS-DA score plot of metabolomic dataset, positive mode (R2Y =0.997 Q2Y = 0.575) (e), negative mode (R2Y =0.981 Q2Y = 0.732) (f); LC-MS analysis of hypertensive rat stool (g-l) samples, PCA score plot distribution of W, LN + SR37-3 and SR61-2 groups in positive mode (g) and negative mode (h); arrangement of OPLS-DA model between metabolomic datasets W and LN, positive mode (R2 = (3245 zxft); Q2= (990, -0.52)) = (i), negative mode (R2 = (32) zxft 3562)) (g-0.945; FIG. Q2Y =0, Q2= (0, Q2Y = 3); FIG. 0, Q2Y = 3).

FIG. 4 is a differential metabolite pathway enrichment, a and e are hierarchical aggregation heatmaps between the W and LN groups constructed from important metabolites from serum and cecal samples, respectively, showing relative increase/decrease in metabolite content and similarity between individual samples; columns correspond to different groups and rows to altered metabolites; color indicates metabolite expression value; green: lowest, red: highest; b. c and d are metabolomic views of serum samples in pathway analysis using metabolic analysis between W and LN, LN and LN + SR37-3, and LN + SR61-2, respectively, while f, g, and h are metabolomic views of cecal content samples.

FIG. 5 is a graph showing the results of histomorphometry of rat kidney in example 3, wherein a is W group (control group), b is LN group (model group), c is LN + SR37-3 group, and d is LN + SR61-2 group.

FIG. 6 is a diagram showing the hemolysis experiment of the strain, wherein a is Lactobacillus plantarum SR37-3,b and Lactobacillus plantarum SR61-2,c are positive controls

FIG. 7 is a standard curve, a standard curve graph (a) and an original curve graph (b) of the fluorescent quantitative PCR detection of the high-efficiency pressure-reducing strain SR 37-3.

FIG. 8 is a standard curve, a standard curve graph (a) and an original curve graph (b) of the fluorescent quantitative PCR detection of the high-efficiency pressure-reducing strain SR61-2.

Detailed Description

In order that the invention may be more readily understood, reference will now be made to the following description taken in conjunction with the accompanying drawings.

Example 1 isolation and characterization of Lactobacillus plantarum SR37-3 and SR61-2

1.1 isolation of Probiotics and seed stock preservation

Collecting Sri Lanka fermented food as sample, adding 0.1g salted fish sample into 10ml MRS liquid culture medium under aseptic environment, shaking, mixing, performing enrichment culture at 37 deg.C under anaerobic condition for 24 hr, and sucking 0.5ml bacteriaThe solution is diluted in a gradient way. Adding physiological saline to obtain 10 ^-1 To 10 ^-5 Diluting the gradiental bacteria suspension, selecting 10 ^-3 、10 ^-4 、10 ^-5 And (3) sucking 100 mu l of three gradient bacterium suspensions to MRS agar culture medium respectively, smearing the three gradient bacterium suspensions uniformly by using a coating rod, and then culturing the three gradient bacterium suspensions for 48 hours under the anaerobic condition at 37 ℃. Selecting typical colony on the plate to MRS agar culture medium, streaking and purifying, selecting single colony after purification, inoculating into MRS liquid culture medium, anaerobically culturing at 37 deg.C for 48h, and storing 30% glycerol in-80 deg.C ultra-low temperature refrigerator.

1.2 identification of Lactobacillus

Bacterial DNA extraction was performed using a bacterial DNA extraction kit (Mabio, CHINA), followed by PCR amplification using 2 × PCR mix (Dongshengbio, CHINA). The PCR amplification primer adopts a 16S rRNA gene universal primer, and the sequence of an upstream primer is 27F:5'-AGA GTT TGA TCC TGG CTC AG-3'; the sequence of the downstream primer is 1492R:5'-CTAC GGC TAC CTT GTTACGA-3'. The PCR reaction conditions are as follows: pre-denaturation at 95 ℃ for 5min; 35 cycles of 95 ℃ 30s,56 ℃ 30s and 72 ℃ 1min 30s, and annealing and extending for 10min at 72 ℃. The PCR product was recovered by cutting and then subjected to one-generation sequencing (performed by Sovia Jin Weizhi Biotech, inc.). The obtained 16S rRNA gene sequences were compared with NCBI database (https:// blast. NCBI. Nlm. Nih. Gov), and for the strains whose Identity and Coverage were more than 99% similar to known Lactobacillus plantarum in the comparison results, it was determined that the strains were Lactobacillus plantarum.

Wherein the gene sequence of the 16S rRNA of the strain claimed by the patent is shown as SEQ ID No. 7-8. The sequences were compared with NCBI database (https:// blast.ncbi.nlm.nih.gov), and the results suggest that they have the highest homology with Lactobacillus plantarum, and they were named Lactobacillus plantarum (Lactobacillus fermentum) SR37-3 (16S rRNA corresponds to SEQ ID No. 7) and SR61-2 (16S rRNA corresponds to SEQ ID No. 8).

Example 2 Whole genome sequencing of two strains of Lactobacillus plantarum

a determination of genomic DNA concentration

1) Extracting strain DNA by using a bacterial DNA extraction kit, and extracting strain DNA

For dsDNAHS regen

The dsDNAHS Buffer was diluted 200-fold and the final volume of assay solution per tube was 200. Mu.L.

2) Adding 190 μ L of the working solution into a standard substance measuring tube, adding 10 μ L of the standard substance, and mixing for 2-3s. 199 microlitres of the working solution is added into each sample measuring tube, 1 microlitres of the sample is added, and after the preparation is finished, the tube wall is flicked by fingers for two to three seconds to be uniformly mixed. The Qubit samples were measured after 2min of storage in the dark.

b fragmentation of genomic DNA

100ng of DNA was added to the sample tube and adjusted to 79. Mu.L with sterile water. DNA was disrupted using Covaris M220 with a size of 500bp

c end repair and purification of fragmented DNA

The sample was transferred from the sample tube to a 1.5mL centrifuge tube and added as per Table 1

TABLE 1 DNAREPIRAGENTS

Incubate at room temperature for 20min and add 180. Mu.L of magnetic beads to the tube. And (3) blowing and beating for 5 times by using a pipette gun to fully and uniformly mix the DNA and the magnetic beads, centrifuging for a short time, and incubating for 5min at room temperature. The sample was placed on a magnetic stand until the solution became clear, and the supernatant was carefully aspirated off with a pipette gun to avoid damaging the magnetic bead layer. Add 500. Mu.L of freshly prepared 70% ethanol to the centrifuge tube, incubate for 30s, rotate the centrifuge tube 2-3 cycles in place. After the solution became clear, the supernatant was carefully aspirated off with a pipette gun to avoid damaging the magnetic bead layer. This step was repeated once. And taking the sample out of the magnetic frame, centrifuging for a short time, putting the sample back on the magnetic frame, and absorbing and discarding residual ethanol to avoid damaging the magnetic bead layer. And opening a tube cover on the magnetic frame to naturally dry the magnetic bead layer at room temperature, wherein the operation process is not more than 5min. The sample was removed from the magnetic stand, 25. Mu.L of sterile water was added, 5 times by pipetting with a pipette, and vortexed for 10 seconds to thoroughly mix the sample. After brief centrifugation, the magnetic frame was returned until the solution became clear, and the supernatant was collected and transferred to a new 0.2mL PCR tube.

d-linker ligation, gap repair and library purification

The corresponding reagents were added to the above sample tubes as in table 2 and centrifuged briefly. The sample tube was then placed in a PCR instrument, programmed as shown in Table 3, and after the reaction was complete the sample was transferred to a new 1.5mL centrifuge tube in preparation for further purification.

TABLE 2 PCR reaction System

TABLE 3 PCR procedure

Add 100. Mu.L of magnetic beads to the tube. And blowing and beating the DNA by a pipette gun to fully and uniformly mix the DNA with the magnetic beads, centrifuging for a short time, and incubating for 5min at room temperature. And (3) placing the sample on a magnetic frame, standing for 3min or carefully absorbing and discarding the supernatant by using a pipette after the solution becomes clear so as to avoid damaging the magnetic bead layer. Add 500. Mu.L of freshly prepared 70% ethanol directly to the centrifuge tube, incubate for 30s, rotate the centrifuge tube 2-3 cycles in situ. After the solution became clear, the supernatant was discarded by pipetting to avoid damaging the magnetic bead layer. And taking the sample out of the magnetic frame, centrifuging for a short time, putting the sample back on the magnetic frame, and absorbing and discarding residual ethanol by using a pipette gun to avoid damaging the magnetic bead layer. And opening a tube cover of the centrifugal tube on the magnetic frame, and naturally airing the magnetic bead layer for 5min at room temperature. The sample was removed from the magnetic stand, 20 μ L of sterile water was added, blown down 5 times with a pipette, vortexed and shaken for 10s to mix well for a short period of time, then placed back on the magnetic stand until the solution became clear and the supernatant collected and transferred to a new 0.2ml PCR tube.

e size screening of linker-containing libraries

Handle

SizeSelect ^TM 2% two combs on the agar Gel were pulled off and inserted into the iBase from left to right ^TM In (1). mu.L of the prepared DNA sample was added to the well (upper row), 10. Mu.L of 50bp DNA Ladder was added to the well M, 25. Mu.L of sterile Water was added to the well (lower row), and 10. Mu.L of nucleic-free Water was added to the well M. Cover the transilluminator, plug the instrument power supply, select SizeSelect in mode ^TM 2% program, setting time 20min. When the indicator band on Ladder, which corresponds to the size of the target recovery band, runs to the reference line, the instrument is halted, about 10. Mu.L of nucleic-free Water is added to the well, the instrument is started, the corresponding band in Ladder is waited to enter the well, the instrument is stopped, and the sample is carefully pipetted into a new 0.2mL PCR tube using a pipette.

f library amplification and purification

The corresponding reagents were added to the sample tubes as shown in table 4. After being fully mixed, the sample tube is put into a PCR instrument

TABLE 4 PCR reaction procedure

The procedure was set up according to the conditions of table 5. After the reaction was complete, transfer the PCR product to a new 1.5mL centrifuge tube.

TABLE 5 PCR reaction procedure

Add 130. Mu.L of magnetic beads to the tube. The DNA was mixed well with the beads, centrifuged briefly and incubated at room temperature for 5min. The sample was placed on a magnetic stand until the solution became clear and the supernatant was discarded by aspiration. Add 500. Mu.L of freshly prepared 70% ethanol to the centrifuge tube, incubate for 30s, rotate the centrifuge tube 2-3 cycles in place. And after the solution becomes clear, sucking and discarding the supernatant, taking the sample out of the magnetic frame, centrifuging for a short time, putting the sample back on the magnetic frame, sucking and discarding residual ethanol by using a 20 mu L pipette gun to avoid damaging the magnetic bead layer, opening a tube cover of a centrifugal tube, naturally airing the magnetic bead layer for 5min at room temperature, taking the sample out of the magnetic frame, adding 20 mu L sterile water, and performing vortex oscillation for 10s to thoroughly mix the sample. After brief centrifugation, the magnetic stand was returned until the solution became clear, and the supernatant was collected and transferred to a new 0.2ml PCR tube.

g determination of library concentration after amplification

Determination of library concentration after amplification and determination of genomic DNA concentration

h carrying out on-machine test

i, sequencing to obtain the complete genome sequences of the lactobacillus plantarum SR37-3 and SR61-2. And comparing VFDB (viral Factor Data base), ARG-anti (inflammatory Resistance Gene-ANNOTAT), CARD (the Comprehensive antigenic Research Database) and Resfinder databases by using Abricite software, and annotating Virulence genes or drug Resistance genes of the databases, wherein the results are shown in tables 6-7. Both strains do not contain virulence genes, and the existence of the genes is determined only if the coverage rate of the drug-resistant genes is more than or equal to 95 percent and the identification rate is more than or equal to 90 percent. The safety of the strain is fully proved by carrying out drug resistance and virulence gene analysis on the strain from the genome level.

TABLE 6 Whole genome virulence and resistance gene analysis of Strain SR37-3 ("/" indicates none)

TABLE 7 Whole genome virulence and resistance genes of Strain SR61-2 ("/" indicates none)

EXAMPLE 3 determination of angiotensin-converting enzyme inhibitory Rate

Inoculating the activated strain into 11% (w/v) skim milk sterilized at 105 deg.C for 15min at an inoculation amount of 4% by volume, and culturing at 37 deg.C until the skim milk coagulates. Centrifuging the prepared fermented milk at 4 ℃ at 7000 Xg for 10min, taking the supernatant, adjusting the pH value of the supernatant to 7.5,4 ℃ by 5mol/L NaOH solution, centrifuging at 11000 Xg for 3min, taking the supernatant, filtering by a 0.45-micron filter membrane, and storing the obtained whey at 4 ℃ for later use. 10 μ L of 0.25U ACE,10 μ L whey samples were added to 4 wells of a 96-well plate, respectively; 10 μ L whey sample, 10 μ L50 mmol/L Tris-HCl;10 μ L of 0.25U ACE,10 μ L of 50mmol/L Tris-HCl;20 μ L50 mmol/L Tris-HCl. 150 μ L of 0.88mmol/L FAPGG in 37 ℃ water bath for 15min was added at the fastest rate. Immediately after the sample addition, the initial absorbance of each sample well was measured at 37 ℃ at 340nm with a microplate reader after shaking for 30s, and each absorbance was designated as a ₁ ，b ₁ ，c ₁ ，d ₁ After incubation at 37 ℃ for 15min, the absorbance of each well was measured again under the same conditions, and each well was designated as a ₂ ，b ₂ ，c ₂ ，d ₂ The absorbance decrease values of the respective sample wells were A = a ₁ -a ₂ 、B＝b ₁ -b ₂ 、C＝c ₁ -c ₂ 、D＝d ₁ -d ₂ . And calculating the ACE inhibition ratio of the fermented milk according to a formula. And calculating to obtain the strain with higher ACE inhibition rate.

ACE inhibition (%) = [ (C-D) - (A-B) ]/(C-D) × 100%

The ACE inhibition rate of the two strains is determined, and the experimental result shows that the inhibition rates of fermented milk supernatants of the lactobacillus plantarum SR37-3 and SR61-2 are 70.5% and 68.9% respectively.

Example 4 in vivo hypotensive Effect of two Lactobacillus plantarum strains and related mechanisms

S1 Experimental design

Wistar rats (8 weeks old) were randomized 7 days after acclimation and were treated as follows:

1. normal group (W): the stomach is irrigated with 10ml/Kg of normal saline every day, and water is freely drunk. 2. Model set (LN): adding 400mg/LN into drinking water ^G -nitro-L-arginine methyl ester hydrochloride (L-NAME), gavage 10ml/Kg of normal saline daily. SR37-3 intervention group (LN + SR 37-3): adding 400mg/L L-NAME into drinking water, and performing intragastric administration 10ml/Kg per daySR37-3 fermented milk (preparation method same as example 2). 4.3.SR61-2 intervention group (LN + SR 61-2): 400mg/L L-NAME is added to the drinking water, and 10ml/Kg SR61-2 fermented milk is gavaged every day (the preparation method is the same as that of example 3). The experiment was approved by the ethical committee and approved as a gavage cycle of 4 weeks, initial systolic and diastolic pressures were measured, and thereafter systolic and diastolic pressures were measured every two weeks, with the blood pressure results shown in fig. 1. The interference of the strains SR37-3 and SR61-2 fermented milk inhibits the increase of the blood pressure. At the end of 4 weeks, long-term administration of probiotic fermented milk inhibited SBP (170.22 + -8.40 and 133.28 + -6.09 for LN + SR37-3, respectively, p + SR37-3 compared to the model group<0.01 P and DBP (LN and LN + SR37-3 are 133.83 + -5.91 and 103.00 + -6.41 respectively, LN + SR37-3 compare with the model group<0.01 Increase in). Two weeks after intake of L-NAME in drinking water, blood pressure (SBP/DBP) increased significantly from 125.67 + -4.26/89.83 + -6.01 mmHg to 157.89 + -6.69/114.22 + -2.55 mmHg. Our results are superior to TSAI et al, which reduces SBP by 19mmHg after gavage of fermented whey into SHR rats. In general, the research results of the people have certain advantages, and different models are adopted for verification in the later period.

Detection of related substances in S2 serum

Animals were fasted for 16h and blood was collected, and serum was separated and then tested using the kit for the following criteria. The results of the detection are shown in FIG. 2. The detection indexes are as follows:

1. relating to the blood pressure regulation mechanism (NE, ET-1, ang I, ang II, ALD)

2. Associated with oxidative stress and inflammation (IFN-beta, TNF-alpha, IL-6, IL-1 beta)

The results show that: SR37-3 and SR61-2 fermented milks can reduce factors related to hypertension mechanism regulation, such as NE, ET-1, ang I, ang II and ALD, and simultaneously can reduce inflammatory factors IFN-beta, TNF-alpha, IL-6 and IL-1 beta in the serum of hypertensive rats. This suggests that SR61-2 and SR37-3 may exert hypotensive effects through the angiotensin system and reduction of inflammatory responses.

S3 serum and stool non-targeted metabolomics

Serum sample pretreatment

Adding 200ul of serum into a mixed solution of 1ml of methanol and acetonitrile (methanol: acetonitrile =1: 1v/v), uniformly mixing, performing ultrasonic treatment in ice water for 20min, then freezing at-20 ℃ for 1h, centrifuging to obtain a supernatant (4 ℃,13000r/min,15 min), concentrating by using a vacuum drying oven, adding 50% acetonitrile for redissolution, and centrifuging to obtain 150ul of serum for loading. And 10ul of each tube was mixed as a QC sample.

Fecal sample pretreatment

Weigh (50 ± 5) mg of solid fecal sample, add pre-cooled methanol: acetonitrile: water (4: water (1, v).

LC-MS on-machine analysis

The chromatographic column is ACQUITY UPLC HSS T3 (2.1X 100,1.8 um). Elution was performed in a gradient with a flow rate of 0.3ml/min and a column temperature of 40 ℃. A heating electrospray ion source (HESI) is adopted, the positive electrode voltage of the spray voltage is 3500V, and the negative electrode voltage is 2000V. The capillary temperature was 320 ℃. The sheath gas flow rate was 45arb, and the auxiliary gas flow rate was 8arb (-) or 10arb (+ or-). A positive and negative ion switching acquisition mode is adopted, and the scanning mode is Full Scan/dd-MS2; full MS resolution was set at MS Full Scan 70000FWHM and MS/MS17500FWHM.

And LC-MS data processing, namely importing the acquired LC-MS/MS data raw file into Compound discover 2.0 software for data preprocessing.

Multivariate statistical analysis: the data obtained above were imported into Simca 14.1 software for analysis. The changes in metabolic profiles of the normal and model groups were explored using orthogonal partial least squares discriminant analysis (OPLS-DA) and differential metabolites were found by combining VIP >1 (serum) and VIP >2 (stool) in the graph and t-test (p < 0.05). The R2 and Q2 values are used to describe the reliability of the data model. Statistical analysis: data are expressed in mean + -SD, sigmaplot14.0 statistical software is used for analysis, independent sample t test is adopted for comparison between two groups, one-factor variance analysis is adopted for comparison between multiple groups, and p <0.05 is considered to have statistical difference. Metabolic profiling analysis: the complex data obtained from the four groups of serum and fecal samples were subjected to dimensionality reduction using PCA-X and OPLS-DA.

From the PCA plot scores, we can observe that in positive ion mode, the W, LN + SR37-3 and LN +61-2 groups are in different positions (FIG. 3a, g), however, in negative ion mode, there is no significant separation between the groups (FIG. 3b, h). The OPLS-DA scores in positive and negative ion mode were significantly separated in both W and LN groups, whether serum or cecal content, as shown in figures 3e, f, k and l, respectively. From the distribution trend, we can further confirm that the hypertension model was successfully established using L-NAME. Chemical identification of these characteristic metabolites is achieved from mass fragment patterns and isotope peak ratios by local databases as well as online databases, comprising HMDB (www.HMDB.ca). Biomarker analysis: serum sample results showed that 30 metabolites between W and LN, 21 metabolites between LN and LN + SR37-3, and 28 metabolites between LN and LN + SR61-2 were identified, as shown in Table 8, including amino acids, fatty acids, amide derivatives, and choline derivatives, and that the results of samples of caecal contents showed that 27 metabolites between W and LN, 31 metabolites between LN and LN + SR37-3, and 24 metabolites between LN and LN + SR61-2, including amino acids, fatty acids, amide derivatives, bile derivatives, pyrimidines, and choline derivatives, as shown in Table 9, were followed by a pathway analysis of characteristic metabolites using Metabolyst 4.0 (www.MetaboAnalyst.ca) in order to more fully understand the mechanism of hypertension. Metabolic pathway analysis: differential metabolites were introduced into MetPA (Metabo Analyst 3.0, http:// www.Metaboanalyst.ca) for pathway enrichment analysis. In serum samples, 7 metabolic pathways significantly affected by hypertension were found due to a significant change in 31 metabolites between W and LN (fig. 4 b), of which 3 are amino acid metabolisms, similar to the results in samples of caecal contents, of which the first five are amino acid metabolisms (fig. 4 f). Based on 22 significantly altered metabolites between LN and LN + SR37-3, 5 metabolic pathways significantly affected by Lactobacillus plantarum SR37-3 fermented milk were found (FIG. 4 c), of which 3 are amino acid metabolisms and mainly histidine and pyrimidine metabolisms were involved in the stool samples (FIG. 4 g). Considering that there are 29 kinds between LN and LN + SR61-2The metabolites were significantly altered and 5 metabolic pathways significantly affected by lactobacillus plantarum strain SR61-2 fermented milk were found (fig. 4 d), mainly involved in fatty acid and amino acid metabolism, but in fecal samples, mainly associated with primary bile acid biosynthesis (fig. 4 h). The heat map shows that there was a clear separation between the LN and W groups in the serum (fig. 4 a) and cecal content (fig. 4 e) samples. However, most of the serum and caecal content metabolites had the same trend in the control and model groups. Metabolites are divided into two large clusters (top and bottom). The metabolites of the upward clusters are mainly enriched in the control group, while the metabolites of the downward clusters are mainly enriched in the disease group. In particular, we have found that L-phenylalanine, L- (-) -methionine and L-valine are three different metabolites that are involved in both intestinal and blood metabolism. Phenylalanine, tyrosine and tryptophan biosynthesis and phenylalanine metabolism are common metabolic pathways of serum and cecal contents, and may be involved in the pathogenesis of the disease. In a blood metabolome, SR37-3 fermented milk can increase the content of nicotinamide in vivo, and the nicotinamide has a series of functions of resisting aging, improving hypercholesterolemia, protecting endothelial cells of patients with coronary heart disease, reducing ET-1 secretion and release and the like. SR61-2 fermented milk can increase the content of betaine and arachidonic acid in vivo. Betaine has certain curative effect on coronary heart disease, liver disease, cardiovascular disease and nervous disease, and has the function of regulating and controlling metabolism balance and physiological balance of human body, and the mechanism is that betaine participates in the reaction of methionine and homocysteine. Arachidonic acid has effects of reducing blood glucose, blood lipid and cholesterol. Research shows that arachidonic acid can obviously reduce low-density lipoprotein (LDL-C) and very low-density lipoprotein (VLDL-C) in blood, and increase high-density lipoprotein (HDL-C), which is of great significance to cardiovascular health. Arachidonic acid also has a series of biological effects of relaxing blood vessels, participating in immune regulation, regulating neuroendocrine, promoting cell division and the like.

TABLE 8 serum differential metabolite assignment table based on UPLC-MS/MS technology blank group and model group

TABLE 9 identification table of cecal content differential metabolites based on UPLC-MS/MS technology blank group and model group

R.T.: retention time (min), W: animals not treated with L-NAME received standard chow, LN L-NAME treated rats received standard chow, LN + SR37-3:L-NAME treated rats received Lactobacillus plantarum strain SR37-3 fermented milk gavage, LN + SR61-2:L-NAME treated rats received Lactobacillus plantarum strain SR61-2 fermented milk gavage. HMDB: human metabolome databases.

S4 Kidney morphological Observation

After the rat in example 3 is dissected, the kidney tissue is taken out and subjected to HE staining to observe the tissue morphology and pathological changes, and the specific operation steps are carried out according to the steps of taking materials, fixing, trimming, dehydrating, transparentizing, waxing, embedding, slicing and copying. The results are shown in FIG. 5.

As can be seen from FIG. 5, the kidney tissue of the control group shows that the glomeruli in the cortex are uniformly distributed, the number of cells and matrixes in the glomeruli are uniform, the tubular epithelial cells are round and full, the brush border is regularly arranged, and the medulla is not obviously abnormal; the connective tissue between urinary tubules is the renal interstitium without obvious hyperplasia of the interstitium; no significant inflammatory cell infiltration was seen. In the model group, renal tissue showed cavitary degeneration of arteriolar smooth muscle cells and microscopic vacuoles in the cytoplasm (black arrows) compared to the control group. While no obvious abnormality was seen in the SR37-3 intervention group, a small amount of tubular nucleus enlargement and vacuolation (black arrows) was seen in the SR61-2 intervention group, and no other obvious abnormality was seen. The long-term hypertension can cause chronic damage to the kidney of the rat, and the probiotic dry disease can have a certain relieving effect.

Example 5 evaluation of safety of two strains of Lactobacillus plantarum in vitro

(1) Antibiotic sensitivity test

6 common antibiotics are selected for drug sensitivity test. The selected antibiotics are as follows: chloramphenicol, tetracycline, erythromycin, gentamicin, amoxicillin, and ampicillin. Centrifuging the prepared bacterial liquid at 4000 Xg for 10min, washing the bacterial liquid for 3 times by using a sterile PBS solution, diluting the bacterial liquid, dipping the diluted bacterial liquid by using a sterile cotton swab, and uniformly coating the surface of the whole flat plate. After the drug sensitive paper is pasted, the drug sensitive paper is lightly pressed by using sterile forceps to be firmly pasted with the surface of the culture medium. Each flat plate with the diameter of 90mm is stuck with 4 drug sensitive paper sheets, and the distance from the center of each paper sheet to the center is not less than 24mm. Marking the name of the antibiotic, measuring the diameter of the inhibition zone from the back of the plate by a caliper after culturing for 18h at 37 ℃, and recording the result. Staphylococcus aureus ATCC25923 is used as a quality control strain, and a patent drug-resistant strain 833-1 is used as a control. The results obtained are shown in Table 10 below (note: R is drug resistant, I is intermediate, and S is sensitive).

Resistance of probiotics to antibiotics can limit the use of probiotics. The bacterial resistance results can be judged as three forms of resistance (R), intermediate (I) and sensitivity (S) according to CLSI 2015 antimicrobial susceptibility test criteria. As can be seen from Table 10, SR37-3 and SR61-2 both showed sensitivity to various antibiotics, well demonstrating the safety profile of both strains.

TABLE 10 antibiotic susceptibility of two strains of Lactobacillus plantarum

(2) Hemolysis test

Under aseptic conditions, the activated bacterial liquid is streaked on a blood plate by using a disposable aseptic inoculating loop, cultured for 48 hours at 37 ℃, and the hemolysis phenomenon is observed. Hemolysis can develop three features on blood plates: (1) alpha hemolysis: grass green hemolysis rings appear around colonies, and this bacterium is generally opportunistic. (2) beta hemolysis: a broad, transparent hemolytic ring appears around the colony, and is generally highly pathogenic. (3) gamma hemolysis: no hemolytic rings appeared around the colonies, and the strains were generally nonpathogenic.

As shown in FIG. 6, no hemolytic ring appeared around each colony, so that both Lactobacillus plantarum strains (SR 37-3 and SR 61-2) were not hemolytic and had good safety.

Example 6 evaluation of in vivo safety of two strains of Lactobacillus plantarum

S1 Experimental design

Wistar male and female rats (190-210 g) were acclimatized for 7 days and subjected to acute toxicity test by the maximum tolerated dose method. Fasting was performed for 16h before the experiment, without restriction of drinking water. Normal group: free to eat and drink without treatment. The treatment group used a maximum gavage. The grouping is as follows:

normal group 1: the males did not take food and drink water freely without treatment.

Normal group 2: the female mice had free access to food and water without treatment.

Treatment group 3: by 1X 10 ¹⁵ Live bacteria (SR 37-3) in CFU/Kg.

Treatment group 4: by 1X 10 ¹⁵ Live CFU/Kg. Bw (SR 37-3) was gavaged once in female mice.

Treatment group 5: by 1X 10 ¹⁵ Live bacteria (SR 61-2) in CFU/Kg. Bw are perfused into the male mouse once.

Treatment group 6: with 110 ¹⁵ Live CFU/Kg. Bw (SR 61-2) was gavaged once in female mice.

After gavage, the patients were continuously observed for 14 days, and the onset and end body weights were measured after recording the signs of intoxication and death. At the end of the experiment, weighing the spleen after the blood sampling and the death, and calculating the liver body ratio and the spleen body ratio.

Organ index, data were analyzed using sigmaplot14.0 software, and mean ± SD values were used to represent each group of values, which were examined for significance using One wayaanova. The mark is used when there is a significant difference between each sex-treated group and the normal sex group. The experimental results (table 11) showed no significant difference in organ index between the administered group and the normal group.

TABLE 11 liver and spleen organ coefficients for each group of animals

Routine and biochemical detection of blood after S2 toxicity test

The routine items for detecting blood by a full-automatic blood analyzer for taking whole blood comprise: 22 items in total are white blood cell count, basophil count, neutrophil count, eosinophil count, lymphocyte count, monocyte count, basophil proportion, neutrophil proportion, eosinophil proportion, lymphocyte proportion, monocyte proportion, erythrocyte count, hemoglobin content, mean erythrocyte volume, mean erythrocyte hemoglobin content, mean erythrocyte hemoglobin concentration, erythrocyte distribution width variation coefficient, erythrocyte distribution width standard deviation, hematocrit, platelet count, mean platelet volume, platelet distribution width, and platelet volume.

And taking serum and detecting blood biochemical projects by using a full-automatic biochemical analyzer. The method comprises the following steps: alanine Aminotransferase (ALT), total Protein (TP), albumin (ALB), globulin (GLOB), bilirubin (TBIL), alkaline phosphatase (ALP), glutamyl transpeptidase (γ -GT), urea Nitrogen (BUN), blood Creatinine (CRE), uric Acid (UA), blood Glucose (GLU), total Cholesterol (TC), triglyceride (TG), high-Density lipoprotein Cholesterol (LDL-C), low-Density lipoprotein Cholesterol (LDL-C) 15 items in total. The results of the above two tests are shown in the attached tables 12, 13, 14 and 16. In the table, SR37-3 indicates that the strain SR37-3 was subjected to the gavage treatment, and SR61-2 indicates that the strain SR61-2 was subjected to the gavage treatment. Data were analyzed using sigmaplot14.0 software and groups of values were expressed as mean ± SD values and tested for significance using One wayaanova. The mark is used when there is a significant difference between each sex-treated group and the normal sex group. The experimental results show that there is no significant difference in organ index between each administration group and the normal group.

TABLE 12 blood routine analysis of individual female groups

TABLE 13 blood routine analysis of various male groups

TABLE 14 Biochemical analysis of blood of animals of each group of females

TABLE 15 Biochemical analysis of blood of various male animals

Example 7 specific Gene fragment of Lactobacillus plantarum SR61-2

(1) Excavation of specific molecular targets of different species of lactobacillus plantarum SR61-2

Performing bioinformatics analysis according to a GenBank database and a whole genome DNA sequence self-tested by the team; screening to obtain specific gene segments of the lactobacillus plantarum SR37-3 and SR61-2, wherein the nucleotide sequences of the gene segments are shown in SEQ ID NO. 1-2. Wherein the sequence SEQ ID NO.1 is a SR37-3 strain specific gene fragment, and the sequence SEQ ID NO.2 is a SR61-2 strain specific gene fragment

(2) Primer validity detection

Specific PCR amplification primer sets (comprising a forward primer and a reverse primer) are designed according to the sequence SEQ ID NO.1 in (1), and the sequences of the primer sets are shown in the following table 16.

TABLE 16 specific PCR detection primer set

S1 DNA template preparation: respectively culturing SR37-3 and SR61-2 in MRS liquid culture medium for enrichment, and respectively extracting DNA by using bacterial genome DNA extraction kit to be used as template to be detected;

s2 PCR amplification:

the PCR detection system is as follows:

primer set 1 PCR amplification procedure:

primer set 2 PCR amplification procedure:

s3: and (3) carrying out gel electrophoresis on the PCR amplification product, and observing whether a band exists at the target position at the position of the size of the corresponding product of each primer group. If no band appears at the desired position for other strain templates, the corresponding target is a strain-specific molecular target.

PCR detection was carried out according to the method of example 6. Wherein, the S1 DNA template is prepared by respectively extracting the genome DNA of each bacterium; and in S2 PCR amplification, the primers used are the primers in the primer group. A blank was set, the template of which was an aqueous solution without genome. The strains of the respective bacteria used and the results of the detection are shown in tables 17 to 18, in which "-" in the columns of the results of the detection indicates negativity.

TABLE 17 evaluation test results of the detection specificity of Lactobacillus plantarum SR37-3 of the present invention

TABLE 18 evaluation test results of detection specificity of Lactobacillus plantarum SR61-2 of the present invention

As can be seen from the above two tables, only the target strain will have an amplified band at the target fragment position and the brightness is high, which indicates that only the target strain in the method contains the specific molecular target.

Example 7 establishment of method for fluorescent quantitative PCR detection of Lactobacillus plantarum SR37-3 and SR61-2

Bacterial DNA was extracted using a genomic DNA extraction kit (magenta Biotech, china) and its concentration was measured. The concentration is converted into copy number, and the conversion formula between the DNA concentration and the copy number is as follows:

wherein A-number of copies per μ LDNA; C-DNA mass concentration, in ng/. Mu.L; l-the length of the DNA fragment, in bp. The DNA was diluted in a gradient so that the copy number was 10 ² ～10 ¹⁰ In the meantime. And performing fluorescent quantitative PCR by using the product after gradient dilution as a template. Fluorescent quantitative PCR system (20 μ L): 2 XSYBR PCR premix 10. Mu.L, forward primer 0.6. Mu.L, reverse primer 0.6. Mu.L, template 1. Mu.L, 50 XRO × Reference Dye 0.4. Mu.L, ddH ₂ O7.4. Mu.L. PCR procedure 1: pre-denaturation at 95 ℃ for 15min, denaturation at 95 ℃ for 10s, annealing at 60 ℃ for 20s, extension at 72 ℃ for 30s, and circulating for 40 times in 2-4 steps. A standard curve was prepared with copy number and Ct value of the corresponding sample. As shown in FIGS. 7 to 8, the Ct value is linear with the lg copy number, and the SR37-3 standard curve is Y =44.089-2.8593X. X represents lg copy number and Y represents Ct value. R2= 0.991. The R61-2 standard curve is Y =45.7602-3.5494X. X represents lg copy number and Y represents Ct value. R2=0.9956.

SEQ ID NO.1

ATGACAACAACATTTGATGACTTGATGGCGTCAATTGACAAAGCAGGTAAAGACGTTCAACGAGATCGAGGCAATATGTTTGAAGATGTGGTTAAGTCCTATCTGACCAATGAGCCTACATACAAACGGGAATTTGACCAAGTCTGGTTATTGTGA

SEQ ID NO.2

ATGGATAATAACTCTAATAATATTACTTTACATATCTTTCCAGATACTGGTGCCATGTGGGAAGGATTAGGCGGAAATTTTGATAATTTTACGGAAATTTTGAACGAATTTATTGATAATGCTATTTCTAATTTACTAAATGTGGAGAATCCTATTAAAAATATTTTAGTTTCTGTTGAACAATCAGTTGATTGTTCAACAGAATATAGGATAACTATAGAAGATACAGGATCAGGAATTCAAAATCTTGAGTCAGCTTTCACTATTGGTAATAAAAGTGGACAAGAATCTCCACTTAATGAACATGGTTATGGTATGAAGCACGCCTTGGCAGCCGCAAATAAAGCTAATGATTCTTGGAAAATATATTCTAAAAAGCAAGAAGATAGCTTTTATTCACTTGTTGCTGCTCCATATTCTTTAAAGAATCAACAAGTTAAAAGAATTAGCAAAAATTTACCTGGACAAATATGTAATACGCATGGCACGATAATAAGTTTTAATATATCTTTTGAATGGCTAAAAACTATTACTAAGGGATTGAGAGGCAATTATACCAAGTTATCTAAATTCATGAATATTTTACAAGAAGATTTAGGGTACACCTATGGTCCATTTTTTTCAGAAAGTGGAATAACTTTAACATTAAGATATAAGGACAATGAATCTAATAGAACTCAAACCCTTAACGTAGAGGAAGTTAAACCTTCAGAAGTTAGAACTTTAAACCCAGGTTCCGGCACAACTACTTATGATTTAAGTAGGGGGCCTATATCCATTGATTATAGGTTTCTTCAAATAAAAAAATCTGAAAAATATGAAAAACATTATTTAGCGAATATGAGTACTTCTGGTGTTGAAATACGTATCAACGGACGTTTGCTAGCTGACAATATTTTCTCAGATATTTGGGGAATTGATAGACACAATTCATATAATTATCTATTGATTAAAATAAATTTAAAATCGACTGACTTGAAAAGATTACCCAAAACCACTACTAATAAGACATCATTAAAGCAAGATGACCCATGCTTGGATAAATTATATTCTTGGATTCGATCAAAAATGCCTACTCCCAAAAAAGAGGTTTCACTTGCAGATGATGAAATAGACCTCTTTTCCGAATTAGAAAAACAAAAAAAGTCATCTTATAAAACAATAGATCCATCGGCCTTAATTACAACCGAACAAAATACATTTTACACATTAGGAGAAAAAATACGTATAGATTTGTATCAATCGGTATTAAATAAGATTGTAATCTACGAAGGAAAGAAAGAAGAAACCCACCCAAAAGATGTCTATCAGCTACTAATGTATTGGGACGGTCTAGTTTACGATGGGGTTCCTGTTGCTGAGGGGATTTTAATTGCCGCTAAACATCCAAACAGTGTTAGAAAAATTGTTGCTCTAAAGAACAATTCTAAAGATGATAACGGAAATATGTATAAAATAACTTTAGAAACATGGAAAGATGAAAATATTGATTATCCTGAATAG

SEQ ID NO.3

GTTCAACGAGATCGAGGCAAT

SEQ ID NO.4

ACCAGACTTGGTCAAATTCCCG

SEQ ID NO.5

CAACGGACGTTTGCTAGCTG

SEQ ID NO.6

GCAACAGGAACCCCATCGTA

SEQ ID NO.7

TACCCCACCGACTTTGGGTGTTACAAACTCTCATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCGACTTCATGTAGGCGAGTTGCAGCCTACAATCCGAACTGAGAATGGCTTTAAGAGATTAGCTTACTCTCGCGAGTTCGCAACTCGTTGTACCATCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCACCAGAGTGCCCAACTTAATGCTGGCAACTGATAATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTATCCATGTCCCCGAAGGGAACGTCTAATCTCTTAGATTTGCATAGTATGTCAAGACCTGGTAAGGTTCTTCGCGTAGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAGCCTTGCGGCCGTACTCCCCAGGCGGAATGCTTAATGCGTTAGCTGCAGCACTGAAGGGCGGAAACCCTCCAACACTTAGCATTCATCGTTTACGGTATGGACTACCAGGGTATCTAATCCTGTTTGCTACCCATACTTTCGAGCCTCAGCGTCAGTTACAGACCAGACAGCCGCCTTCGCCACTGGTGTTCTTCCATATATCTACGCATTTCACCGCTACACATGGAGTTCCACTGTCCTCTTCTGCACTCAAGTTTCCCAGTTTCCGATGCACTTCTTCGGTTGAGCCGAAGGCTTTCACATCAGACTTAAAAAACCGCCTGCGCTCGCTTTACGCCCAATAAATCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAAATACCGTCAATACCTGAACAGTTACTCTCAGATATGTTCTTCTTTAACAACAGAGTTTTACGAGCCGAAACCCTTCTTCACTCACGCGGCGTTGCTCCATCAGACTTTCGTCCATTGTGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTTTGGGCCGTGTCTCAGTCCCAATGTGGCCGATTACCCTCTCAGGTCGGCTACGTATCATTGCCATGGTGAGCCGTTACCCCACCATCTAGCTAATACGCCGCGGGACCATCCAAAAGTGATAGCCGAAGCCATCTTTCAAGCTCGGACCATGCGGTCCAAGTTGTTATGCGGTATTAGCATCTGTTTCCAGGTGTTATCCCCCGCTTCTGGGCAGGTTTCCCACGTGTTACTCACCAGTTCGCCACTCACTCAAATGTAAATCATGATGCAAGC

SEQ ID NO.8

TACCCCACCGACTTTGGGTGTTACAAACTCTCATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCGACTTCATGTAGGCGAGTTGCAGCCTACAATCCGAACTGAGAATGGCTTTAAGAGATTAGCTTACTCTCGCGAGTTCGCAACTCGTTGTACCATCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCACCAGAGTGCCCAACTTAATGCTGGCAACTGATAATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTATCCATGTCCCCGAAGGGAACGTCTAATCTCTTAGATTTGCATAGTATGTCAAGACCTGGTAAGGTTCTTCGCGTAGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAGCCTTGCGGCCGTACTCCCCAGGCGGAATGCTTAATGCGTTAGCTGCAGCACTGAAGGGCGGAAACCCTCCAACACTTAGCATTCATCGTTTACGGTATGGACTACCAGGGTATCTAATCCTGTTTGCTACCCATACTTTCGAGCCTCAGCGTCAGTTACAGACCAGACAGCCGCCTTCGCCACTGGTGTTCTTCCATATATCTACGCATTTCACCGCTACACATGGAGTTCCACTGTCCTCTTCTGCACTCAAGTTTCCCAGTTTCCGATGCACTTCTTCGGTTGAGCCGAAGGCTTTCACATCAGACTTAAAAAACCGCCTGCGCTCGCTTTACGCCCAATAAATCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAAATACCGTCAATACCTGAACAGTTACTCTCAGATATGTTCTTCTTTAACAACAGAGTTTTACGAGCCGAAACCCTTCTTCACTCACGCGGCGTTGCTCCATCAGACTTTCGTCCATTGTGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTTTGGGCCGTGTCTCAGTCCCAATGTGGCCGATTACCCTCTCAGGTCGGCTACGTATCATTGCCATGGTGAGCCGTTACCCCACCATCTAGCTAATACGCCGCGGGACCATCCAAAAGTGATAGCCGAAGCCATCTTTCAAGCTCGGACCATGCGGTCCAAGTTGTTATGCGGTATTAGCATCTGTTTCCAGGTGTTATCCCCCGCTTCTGGGCAGGTTTCCCACGTGTTACTCACCAGTTCGCCACTCACTCAAATGTAAATCATGATGCAAGCACCAATCAATACCAGAGTTCGTTCGACTGC。

Claims (10)

1. Lactobacillus plantarum (Lactobacillus plantarum) SR37-3 with the deposit number: GDMCCNo:62390.

2. lactobacillus plantarum (Lactobacillus plantarum) SR61-2, deposit number is: GDMCCNo:62389.

3. use of lactobacillus plantarum SR37-3 according to claim 1 or lactobacillus plantarum SR61-2 according to claim 2 for the preparation of a product for lowering blood pressure, reducing serum production of inflammatory substances, improving kidney damage and promoting cardiovascular health.

4. Use according to claim 3, wherein the product is a pharmaceutical or functional food.

5. The use according to claim 4, wherein the functional food further comprises an adjuvant, inulin, resistant starch, fructo-oligosaccharide or galacto-oligosaccharide.

6. Use according to claim 4, wherein the functional food is a fermented food product.

7. A product for lowering blood pressure, alleviating serum inflammatory substances, improving kidney damage and promoting cardiovascular health, which comprises the Lactobacillus plantarum SR37-3 of claim 1 or the Lactobacillus plantarum SR61-2 of claim 2 as an active ingredient.

8. The lactobacillus plantarum SR37-3 according to claim 1 or the lactobacillus plantarum SR61-2 according to claim 2, wherein the specific nucleotide sequence of the lactobacillus plantarum SR37-3 is represented by SEQ ID No.1, and the specific nucleotide sequence of the lactobacillus plantarum SR61-2 is represented by SEQ ID No. 2.

9. The primer group for detecting lactobacillus plantarum SR37-3 of claim 1 or lactobacillus plantarum SR61-2 of claim 2, wherein the primer group for detecting lactobacillus plantarum SR37-3 comprises nucleotide sequences shown as SEQ ID nos. 3 and 4, and the nucleotide sequence for detecting lactobacillus plantarum SR61-2 is shown as SEQ ID nos. 5 and 6.

10. A method for detecting Lactobacillus plantarum SR37-3 according to claim 1 or Lactobacillus plantarum SR61-2 according to claim 2, comprising the steps of:

s1: performing PCR amplification using the primer set of claim 9;

s2: carrying out gel electrophoresis to detect the amplification product;

s3: observing whether the amplification product is in accordance with the expectation.

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