Application of pediococcus acidilactici ZJUIDS13 in preparation of medicine for improving alcoholic cardiomyopathy

Abstract

The invention relates to the technical field of food microorganisms, in particular to application of pediococcus acidilactici ZJUIDS13 in improvement of alcoholic cardiomyopathy, and discloses a preparation method of the pediococcus acidilactici ZJUIDS13. The pediococcus acidilactici ZJUIDS13 obviously reduces the plasma CK-MB content in an alcoholic cardiomyopathy model, recovers the cardiac ejection capacity in the alcoholic cardiomyopathy, relieves myocardial hypertrophy, and obviously reduces myocardial damage caused by the alcoholic cardiomyopathy model.

Description

Technical Field

[0001] The invention relates to the technical field of food microorganisms, and in particular to application of Pediococcus acidilactici ZJUIDS13 in improving alcoholic cardiomyopathy. Background Art

[0002] Alcoholic cardiomyopathy (ACM) is a serious heart disease caused by long-term, heavy drinking. Its pathological features mainly include damaged myocardial cells, cardiac enlargement, and decreased cardiac pumping function. Studies have shown that heavy drinking can induce oxidative stress, inflammatory response, and myocardial cell death, which in turn affects cardiac function. Although quitting drinking is a key measure to treat this disease, many patients may experience discomfort after quitting drinking, incomplete recovery of heart function, and multiple risks such as recurrence of the disease. Therefore, it is of great significance to study safe and effective treatments to improve alcoholic cardiomyopathy.

[0003] In recent years, the application value of probiotics in disease prevention and treatment has increased significantly.

[0004] The invention of CN109182225A, "A strain of Pediococcus acidilactici and its application in anti-atherosclerosis", informs that: Pediococcus acidilactici AS185 has the function of improving the serum lipid level of rats with atherosclerosis model, and can significantly reduce the level of adhesion molecules in the serum of rats with atherosclerosis model, thereby preventing the occurrence of atherosclerosis; specifically: reducing total cholesterol (TC) and total triglycerides (TG), increasing high-density lipoprotein cholesterol (HDL-C); reducing low-density lipoprotein cholesterol (LDL-C), reducing intercellular adhesion molecule-1 (ICAM-1), vascular endothelial cell adhesion molecule-1 (VCAM-1). Pediococcus acidilactici AS185 has an inhibitory effect on inflammatory factors such as interleukin (IL-6), tumor necrosis factor α (TNF-α), and gamma interferon (IFN-γ) in the pathological changes of atherosclerosis, and has a promoting effect on the anti-inflammatory factor interleukin 10 (IL-10), thereby inhibiting or slowing the formation of atherosclerosis. Pediococcus acidilactici AS185 can inhibit the upregulation of p38MAPK and pERK1 / 2MAPK signaling pathways caused by a high-fat diet, indicating that Pediococcus acidilactici AS185 may prevent the formation of atherosclerosis in rats by inhibiting the p38MAPK and ERK MAPK signaling pathways.

[0005] As a type of lactic acid bacteria, Pediococcus acidilactici ZJUIDS13 has become one of the research hotspots due to its excellent probiotic properties and wide range of health benefits. The invention of 202211133365.2, "Pediococcus acidilactici ZJUIDS13 with the effect of relieving ulcerative colitis and its application", informs that Pediococcus acidilactici ZJUIDS13 (CGMCC NO.25529) has the effect of relieving ulcerative colitis. Summary of the invention

[0006] The technical problem to be solved by the present invention is to provide application of Pediococcus acidilactici ZJUIDS13 in improving alcoholic cardiomyopathy.

[0007] In order to solve the above technical problems, the present invention provides the use of Pediococcus acidilactici ZJUIDS13 (CGMCC NO.25529) in the preparation of a drug for improving alcoholic cardiomyopathy.

[0008] As an improvement of the application of the present invention: protecting alcohol-induced myocardial damage.

[0009] As a further improvement of the application of the present invention, at least any of the following is included:

[0010] Reduce the level of creatine kinase isoenzyme (CK-MB) in serum;

[0011] Restore myocardial capacity (myocardial ejection capacity);

[0012] Avoid alcohol-induced myocardial hypertrophy.

[0013] Alleviate myocardial damage (including increasing Bcl2 mRNA and decreasing Bax mRNA).

[0014] As a further improvement of the application of the present invention: comprising Pediococcus acidilactici ZJUIDS13 and inactivated Pediococcus acidilactici ZJUIDS13.

[0015] As a further improvement of the application of the present invention, it includes a suspension prepared by using the bacterial powder of Pediococcus acidilactici ZJUIDS13 and an inactivated ZJUIDS13 bacterial liquid.

[0016] The beneficial effects of the present invention are mainly reflected in: the lactic acid Pediococcus ZJUIDS13 of the present invention significantly reduces the plasma CK-MB content in the alcoholic cardiomyopathy model, restores the cardiac ejection capacity in alcoholic cardiomyopathy and reduces myocardial hypertrophy, and significantly reduces the myocardial damage caused by the alcoholic cardiomyopathy model. At present, the clinical manifestations of alcoholic cardiomyopathy are often accompanied by cardiac enlargement and congestive heart failure, among which left ventricular failure is the main one. Lactic acid Pediococcus ZJUIDS13 can effectively restore the increase in the thickness of the posterior wall of the left ventricle and myocardial damage, so lactic acid Pediococcus ZJUIDS13 effectively restores the heart damage of drinking mice. In addition, lactic acid Pediococcus ZJUIDS13 restores the Bcl2 mRNA level in the heart of drinking mice and reduces the Bax mRNA level, which reflects that the resistance of myocardial cells to apoptosis is enhanced, and lactic acid Pediococcus ZJUIDS13 can effectively reduce myocardial damage.

[0017] As a complex heart disease, alcoholic cardiomyopathy urgently needs innovative treatment strategies. This paper deeply studies the potential effects of Pediococcus acidilactici ZJUIDS13 on alcoholic cardiomyopathy, and is expected to provide a more novel, effective and economical treatment option for this intractable disease, thereby improving patient prognosis and reducing the burden of the disease. The dose required for humans can be calculated according to the corresponding dose for mice. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The specific implementation modes of the present invention are further described in detail below with reference to the accompanying drawings.

[0019] Figure 1 The present invention shows the influence of Pediococcus acidilactici ZJUIDS13 on creatine kinase isoenzyme (CK-MB) in mouse plasma.

[0020] Figure 2 The present invention discloses the Pediococcus acidilactici ZJUIDS13 and its effect on the left ventricular ejection fraction of mice.

[0021] Figure 3 The present invention discloses the Pediococcus acidilactici ZJUIDS13 and its effect on the short axis shortening rate of the left ventricle in mice.

[0022] Figure 4 The present invention shows the influence of Pediococcus acidilactici ZJUIDS13 on the thickness of the left ventricular posterior wall in the diastolic period of the mouse heart.

[0023] Figure 5 The invention discloses the effect of Pediococcus acidilactici ZJUIDS13 on the thickness of the posterior wall of the left ventricle during the systolic period in the mouse heart.

[0024] Figure 6 The present invention shows the influence of Pediococcus acidilactici ZJUIDS13 on the heart weight and the ratio of heart to body weight of mice.

[0025] Figure 7 The present invention shows the effect of Pediococcus acidilactici ZJUIDS13 on Bcl2 mRNA in mouse heart.

[0026] Figure 8 The present invention shows the influence of Pediococcus acidilactici ZJUIDS13 on Bax mRNA in mouse heart.

[0027] Fig. 9 The present invention shows the influence of the inactivated Pediococcus acidilactici ZJUIDS13 on creatine kinase isoenzyme (CK-MB) in mouse plasma.

[0028] Fig.10 The invention discloses the inactivated Pediococcus acidilactici ZJUIDS13 and its effect on the left ventricular ejection fraction of mice.

[0029] Fig.11 The invention discloses the inactivated Pediococcus acidilactici ZJUIDS13 and its effect on the short axis shortening rate of the left ventricle in mice.

[0030] Fig.12 The invention discloses the effect of the inactivated Pediococcus acidilactici ZJUIDS13 on the thickness of the left ventricular posterior wall in the diastolic period of the mouse heart.

[0031] Fig.13 The invention discloses the inactivated Pediococcus acidilactici ZJUIDS13 and its effect on the thickness of the posterior wall of the left ventricle during the systolic period in the mouse heart.

[0032] Fig.14 The invention discloses the effects of the inactivated Pediococcus acidilactici ZJUIDS13 on the heart weight and the ratio of heart to body weight of mice.

[0033] Fig.15 The figure shows the effect of the inactivated Pediococcus acidilactici ZJUIDS13 of the present invention on Bcl2 mRNA in mouse heart.

[0034] Fig.16 The figure shows the effect of the inactivated Pediococcus acidilactici ZJUIDS13 of the present invention on Bax mRNA in mouse heart. DETAILED DESCRIPTION

[0035] The present invention is further described below in conjunction with specific embodiments, but the protection scope of the present invention is not limited thereto:

[0036] The following case uses Pediococcus acidilactici ZJUIDS13 (CGMCCNO.25529).

[0037] Example 1, preparation of Pediococcus acidilactici ZJUIDS13 bacterial powder (Pediococcus acidilactici ZJUIDS13 freeze-dried bacterial powder):

[0038] Prepared according to Example 9 of the invention

[0038] "Pediococcus acidilactici ZJUIDS13 with the efficacy of alleviating ulcerative colitis and its application" with the patent number 202211133365.2, and stored at -20°C.

[0039] Example 2. Preparation of heat-inactivated Pediococcus acidilactici ZJUIDS13 bacterial liquid

[0040] Pick a single colony of Pediococcus acidilactici ZJUIDS13 and inoculate it into 50 mL of MRS liquid medium, and place it in an incubator at 37°C for 18 h. Then, activate it again in 250 mL of MRS liquid medium at an inoculation amount of 5% (v / v), and place it in an incubator at 37°C for 24 h. Finally, the activated Pediococcus acidilactici ZJUIDS13 is anaerobically cultured at a high density in a 10 L fermenter at an inoculation amount of 5%, and anaerobically cultured at 37°C and pH 6.8 for 18 h. The bacterial liquid density reaches 10 9 ~10 10 CFU / mL or so, serving as the bacterial liquid (live bacterial liquid).

[0041] Method for preparing heat-inactivated Pediococcus acidilactici ZJUIDS13 bacterial liquid: Heat-inactivate the above-mentioned bacterial liquid of ZJUIDS13 at 121°C under high temperature and high pressure for 30 min to obtain heat-inactivated ZJUIDS13 bacterial liquid.

[0042] Experiment 1. Alleviating effect of Pediococcus acidilactici ZJUIDS13 on alcoholic cardiomyopathy

[0043] 1. Experimental materials:

[0044] Experimental animals: 15 male C57BL / 6 mice, purchased from Shanghai Slack Experimental Animal Co., Ltd., company license number: SCXK (Shanghai) 2017-0005, and raised in the Experimental Animal Center of Zhejiang Chinese Medical University under SPF conditions.

[0045] Reagents: Pediococcus acidilactici ZJUIDS13; choline, vitamins (Nantong Trofi Feed Technology Co., Ltd.); absolute ethanol (CAS-NO: 64-17-5, lichrosolv); creatine kinase isoenzyme (Lot: N28018610, CUSABIO).

[0046] Lieber-DeCarli alcohol liquid model feed (product code TP4030B, Nantong Trofi Feed Technology Co., Ltd.); Lieber-DeCarli control liquid feed (product code TP4030BC, Nantong Trofi Feed Technology Co., Ltd.).

[0047] Animal experiments: Before oral administration, freeze-dried powder of Pediococcus acidilactici ZJUIDS13 was suspended in PBS buffer (0.01 M, pH = 7.2). The number of viable cells in the suspension of Pediococcus acidilactici ZJUIDS13 for oral administration was about 10 9 CFU / mL. The intragastric volume of each mouse was 200 μL. The number of viable bacteria per gram of freeze-dried bacterial powder was counted and corrected on MRS agar plates every week during the experimental period.

[0048] 2. Methods

[0049] Breeding environment: room temperature 20-25℃, light and dark alternation every 12 hours, relative humidity about 50%, feed, drinking water, etc. are uniformly matched, keep the indoor environment well ventilated, clean every day, and maintain a good sanitary environment in the cage.

[0050] After one week of adaptive feeding, C57BL / 6 mice were randomly divided into three groups, namely, pair-fed (PF), alcohol-fed (AF), and alcohol-fed + Pediococcus acidilactici ZJUIDS13 (AF+ZJUIDS13). The mice were fed with Lieber-DeCarli liquid alcohol feed for 4 weeks to establish the mouse model of alcoholic liver disease. In the first three days, mice in each group were adaptively fed with non-alcoholic Lieber-DeCarli control liquid feed, and the daily liquid feed amount for each mouse was 15g. After the adaptive feeding, the AF group and the AF+ZJUIDS13 group were fed with alcohol-containing Lieber-DeCarli liquid feed for 4 weeks, and the PF group continued to be fed with Lieber-DeCarli control liquid feed for 4 weeks. During this period, the control liquid feed amount of the PF group mice was adjusted according to the daily alcohol liquid feed intake of the AF group mice. Since the first day of feeding with liquid feed, the PF and AF groups were gavaged with 200 μL PBS every day, and the AF+ZJUIDS13 group was gavaged with 200 μL ZJUIDS13 suspension.

[0051] The liquid feed consumed by mice was replaced with newly prepared feed at 17:00 every day. The weight of mice was recorded every week to detect changes. After feeding, 10% chloral hydrate was injected intraperitoneally for anesthesia, blood was taken from the apex of the heart to measure creatine kinase isoenzyme (CK-MB), and part of the mouse heart tissue was taken for the measurement of relevant indicators.

[0052] 3. Index determination

[0053] Follow the instructions for use of the corresponding kits, and the components used are provided by the corresponding kits.

[0054] The kits used were as follows: Evo M-MLV reverse transcription premix kit (Cat. No.: AG11728, Aikerui Biotechnology Co., Ltd.), live cell Caspase-3 activity and Annexin V apoptosis detection kit (Cat. No.: C1077S, Beyotime Biotechnology), and creatine kinase isoenzyme kit (Cat. No.: N28018610, CUSABIO).

[0055] The specific operations are as follows:

[0056] 3.1 Plasma CK-MB detection

[0057] Take mouse plasma samples and collect whole blood into tubes containing anticoagulants. EDTA sodium salt is recommended as the anticoagulant. Centrifuge at 1000×g for 15 minutes within 30 minutes after sample collection and take the supernatant. 100 μL of standard working solution and test sample were added to each reaction well. The standard needed to be repeated. After sealing the plate, incubate at 37°C for 90 min. Discard the liquid, spin dry, add 100 μL of biotin-labeled creatine kinase isozyme MB antibody working solution to each reaction well, seal the plate and incubate at 37°C for 60 min. Wash, repeat 4 times. Add 100 μL of HRP-labeled streptavidin working solution to each reaction well, seal the plate and incubate at 37°C for 30 min. Wash, repeat 4 times. Add 90 μL of color developer (protected from light) to each reaction well, seal the plate and develop color at 37°C for about 15 min. Add 50 μL of stop solution to each reaction well, and immediately measure the OD value at 450 nm using an enzyme reader. Calculate the average OD value of the standard and sample. Use the "four-parameter logistics model" to draw a standard curve with the concentration of the standard as the horizontal axis and the absorbance OD value as the vertical axis.

[0058] 3.2 Left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (FS) testing

[0059] Mice were initially anesthetized in a chamber (2% isoflurane, 95% oxygen, 0.8 L / min) and maintained anesthesia (1.5% isoflurane) in supine position on a 37°C heating pad. Echocardiographic assessments were performed using a Vevo 1100 ultrasound device (visualsonic) and a real-time micro-visualization transducer (MS400, 30 MHz) applied to the shaved chest wall.

[0060] Images were acquired in parasternal long axis, parasternal short axis, and short axis M-mode. Switching to M-mode, the M-mode axis was positioned in the middle layer of the left ventricle. Images of cardiac wall thickness, chamber dimensions, and foreshortening were captured for subsequent measurements. Using the “heart package,” SAX (short axis) parameters were selected and measured, including ventricular septum end diastolic (IVSd), ventricular septum end systolic (IVSs), left ventricular end diastolic value (LVDd), left ventricular end systolic value (LVDs), left ventricular posterior wall end diastolic value (LVPWd), left ventricular posterior wall end systolic value (LVPWs). SAMM images were acquired by placing the M-mode cursor vertically on the parasternal short axis view. Views and data were exported for offline calculations using the dedicated Vevo 1100 quantification software. LV Vol-d (μL; left ventricular diastolic volume), LV Vol-s (μL; left ventricular systolic volume), LV EF (%; left ventricular ejection fraction), FS (%; left ventricular fractional shortening) were measured using SAMM images. All procedures were in accordance with the guidelines for mouse cardiac physiological measurements.

[0061] 3.3 Real-time PCR

[0062] (1) Take out the tissue from the -80°C freezer and place it in an ice box. Use an electronic balance to weigh about 0.02 g of tissue into an EP tube and precool the centrifuge at 4°C.

[0063] (2) Add 1 mL of Trizol and 3 steel beads to the EP tube containing the tissue block, grind with a grinder, remove the liquid, pour out the steel beads, and react at room temperature for 10 min;

[0064] (3) Add 200 μL of chloroform to the EP tube, shake vigorously for 30 seconds, and place on ice for 5-10 minutes;

[0065] (4) After standing, place the centrifuge tube in a 4°C centrifuge at 12,000 rpm for 15 min;

[0066] (5) Pipette the aqueous phase (supernatant) of the centrifuged sample into a new 1.5 mL centrifuge tube;

[0067] (6) Add an equal volume of isopropanol solution to the centrifuged solution, gently invert to mix, and let stand at -20°C for 20 min;

[0068] (7) Take out the sample from -20°C and centrifuge it at 12000 rpm and 4°C for 15 min;

[0069] (8) After centrifugation, discard the supernatant to obtain a white (or colorless and transparent) precipitate. Add 100-300 μL of pre-cooled 75% ethanol prepared with DEPC water along the inner wall of the centrifuge tube and wash 2-3 times.

[0070] (9) After discarding the liquid, air-dry at room temperature for about 15 min. Add 20-50 μL of pre-cooled DEPC water to the centrifuge tube to dissolve the air-dried precipitate (RNA) at the bottom, and store in a -20°C refrigerator until use.

[0071] (10) Use an ultra-micro-volume UV spectrophotometer to measure the RNA concentration, record the results, and calculate the sample loading amount for each group.

[0072] (11) Add the corresponding reaction solution in the reverse transcription kit to premix, add the sample for reverse transcription, shake and centrifuge, place in a PCR instrument, and set the corresponding PCR reaction conditions according to the reverse transcription kit. Store the cDNA sample reverse transcribed by the reverse transcription instrument in a -20°C refrigerator for later use.

[0073] (12) Use a 0.2 mL fluorescent quantitative PCR eight-tube concatenation, add the reactants, use cDNA as a template, use a fluorescent quantitative PCR amplification instrument to amplify the experimental target gene, and perform fluorescent quantitative PCR determination.

[0074] (13) The mixed reactants were shaken and mixed, and after centrifugation, the eight-tube strip was placed in a qRT PCR reactor. The PCR reaction program was set as follows: pre-denaturation: 94°C, 5 min; denaturation: 94°C, 30 s, 60°C, 30 s, 72°C, 30 s, 40 cycles; extension: 72°C, 5 min, and storage at 4°C. The hot cover temperature of the PCR instrument was set to 105°C.

[0075] (14) The expression changes of the target genes were calculated using the 2–ΔΔCT method.

[0076] 3.4 Apoptosis detection at mRNA level

[0077] The live cell Caspase-3 activity and Annexin V apoptosis detection kit (Biyuntian Biotechnology Product No.: C1077S) were used.

[0078] Accurately weigh the heart tissue, add lysate at a ratio of 100 μL lysate per 3-10 mg tissue, homogenize with a glass homogenizer on an ice bath; transfer the homogenate to a 1.5 mL centrifuge tube and lyse again on ice for 5 minutes; centrifuge at 16,000-20,000g for 10-15 minutes at 4°C; transfer the supernatant to a centrifuge tube pre-cooled on ice; set up the reaction system, blank control: 40 μL detection buffer, 50 μL lysate, 10 μL Ac-DEVD-pNA (2 mM), sample: 40 μL detection buffer, 50 μL sample, 10 μL Ac-DEVD-pNA (2 mM). Add the detection buffer first, then the sample to be tested, mix appropriately, and be careful to avoid bubbles during mixing. Then add Ac-DEVD-pNA (2 mM); incubate at 37°C for 60-120 minutes. Measure A405 when the color change is obvious. The pNA produced catalytically in the sample was calculated.

[0079] 4. Experimental Results

[0080] like Figure 1 The results showed that the content of creatine kinase isoenzyme (CK-MB) in the serum of mice in the alcohol group (AF group) was significantly higher than that in the control group, and the alcohol group (AF group) had obvious myocardial damage, which means that long-term drinking can cause myocardial damage, and Pediococcus acidilactici ZJUIDS13 can significantly reduce the content of CK-MB and effectively alleviate myocardial damage. Therefore, Pediococcus acidilactici ZJUIDS13 can significantly reduce the increase of CK-MB caused by alcohol intake.

[0081] like Figure 2 and Figure 3 As shown, it can be seen that the left ventricular ejection fraction and left ventricular short-axis shortening rate of the mice in the alcohol group (AF) were significantly lower than those in the control group (PF), while the left ventricular ejection fraction and left ventricular short-axis shortening rate of the mice in the AF+ZJUIDS13 group were significantly increased compared with those in the AF group. Therefore, it can be judged that the myocardial capacity of the mice in the AF group is impaired and the cardiac ejection capacity is reduced, and ZJUIDS13 can effectively restore the myocardial ejection capacity of mice.

[0082] like Figure 4 and Figure 5 As shown, it is the thickness of the left ventricular posterior wall during diastole and systole of mice. It can be seen that the thickness of the left ventricular posterior wall in the hearts of mice in the alcohol group (AF) was significantly increased compared with that in the control group (PF), and there was no significant difference in the thickness of the left ventricular posterior wall in the hearts of mice in the AF+ZJUIDS13 group and the control group (PF). Therefore, it is judged that ZJUIDS13 can effectively reduce the increase in left ventricular wall thickness caused by alcohol in mice and avoid the symptoms of myocardial hypertrophy caused by long-term alcohol intake.

[0083] like Figure 6As shown, it is the heart weight and the ratio of heart to volume of mice. It can be seen that the heart weight of mice in the alcohol group (AF) increased, and the ratio of heart to body weight was higher. The heart weight and heart-to-body ratio of the AF+ZJUIDS13 group were significantly reduced, alleviating the symptoms of myocardial hypertrophy caused by drinking.

[0084] like Figure 7 and Figure 8 As shown, it is the level of Bcl2 mRNA and Bax mRNA in mouse cardiomyocytes. It can be seen that the Bcl2 mRNA in the alcohol group (AF) group was significantly lower than that in the control group (PF), and the Bcl2 mRNA level in the AF+ZJUIDS13 group was significantly restored compared with the alcohol group (AF). In addition, the Bax mRNA in the alcohol group (AF) group was significantly increased compared with the control group (PF), and the Bax mRNA level in the AF+ZJUIDS13 group was significantly reduced compared with the alcohol group (AF). By analyzing the effects of long-term drinking on the levels of Bcl2 mRNA and Bax mRNA, it is shown that ZJUIDS13 can effectively reduce myocardial damage.

[0085] Comparative experiment 1, the lactic acid Pediococcus AS185 was experimented according to the method described in the above experiment 1, and the results obtained were not significantly different from the AF group, so it was shown that lactic acid Pediococcus AS185 could not significantly reduce the increase of CK-MB caused by alcohol intake and alleviate myocardial damage. Lactic acid Pediococcus AS185 failed to improve the decrease of left ventricular ejection fraction and left ventricular short axis shortening rate in the mouse heart caused by alcohol intake, and did not restore the myocardial ejection capacity of drinking mice. Lactic acid Pediococcus AS185 could not effectively reduce the increase of left ventricular wall thickness in mice caused by alcohol, nor could it reduce the heart weight and heart to volume ratio of mice, so lactic acid Pediococcus AS185 could not reduce the symptoms of myocardial hypertrophy caused by drinking. Lactic acid Pediococcus AS185 could not significantly increase Bcl2mRNA and reduce Bax mRNA levels. Therefore, it was proved that lactic acid Pediococcus AS185 could not effectively improve alcoholic cardiomyopathy.

[0086] The results corresponding to the strains of the present invention are: Lactic acid Pediococcus ZJUIDS13 can significantly reduce the increase in CK-MB caused by alcohol intake and effectively reduce myocardial damage. Lactic acid Pediococcus ZJUIDS13 can effectively improve the reduction in left ventricular ejection fraction and left ventricular short axis shortening rate in mice caused by alcohol intake, and effectively restore the myocardial ejection capacity of mice. Lactic acid Pediococcus ZJUIDS13 can effectively reduce the increase in left ventricular wall thickness caused by alcohol in mice, significantly reduce heart weight and heart-body ratio, so the strains of the present invention can improve the symptoms of myocardial hypertrophy caused by alcohol intake. Lactic acid Pediococcus ZJUIDS13 can significantly increase Bcl2 mRNA and reduce Bax mRNA levels, indicating that lactic acid Pediococcus ZJUIDS13 can effectively reduce myocardial damage.

[0087] Experiment 2: The alleviating effect of inactivated Pediococcus acidilactici ZJUIDS13 on alcoholic cardiomyopathy

[0088] In Experiment 1, the 9 CFU / mL ZJUIDS13 suspension" was changed to the "10 CFU / mL ZJUIDS13 suspension" prepared in Example 2 9 CFU / mL of inactivated ZJUIDS13 bacterial liquid", thus forming the alcohol feed feeding group + inactivated Pediococcus acidilactici ZJUIDS13 (AF+ZJUIDS13 (heat-killed)) group.

[0089] That is, the PF and AF groups were intragastrically administered with 200 μL PBS every day, and the (AF+ZJUIDS13 (heat-killed) group) group was intragastrically administered with 200 μL inactivated Pediococcus acidilactici ZJUIDS13 bacterial solution.

[0090] The rest is the same as Experiment 1.

[0091] Experimental results:

[0092] like Fig. 9 The results showed that the serum creatine kinase isoenzyme (CK-MB) content of mice drinking alcohol for a long time was significantly higher than that of mice in the control group, and the alcohol group (AF group) had obvious myocardial damage, which means that long-term drinking can cause myocardial damage. Inactivated Pediococcus acidilactici ZJUIDS13 can significantly reduce the content of CK-MB and effectively alleviate myocardial damage. Therefore, inactivated Pediococcus acidilactici ZJUIDS13 can significantly reduce the increase of CK-MB caused by alcohol intake.

[0093] like Fig.10 and Fig.11 As shown, it can be seen that the left ventricular ejection fraction and left ventricular short-axis shortening rate of the mice in the alcohol group (AF) were significantly lower than those in the control group (PF), while the left ventricular ejection fraction and left ventricular short-axis shortening rate of the mice in the AF+ZJUIDS13 (heat-killed) group were significantly increased compared with those in the AF group. Therefore, it can be judged that the myocardial capacity of the mice in the AF group was impaired and the cardiac ejection capacity was reduced, and ZJUIDS13 can effectively restore the myocardial ejection capacity of the mice.

[0094] like Fig.12 and Fig.13As shown, it is the thickness of the left ventricular posterior wall during diastole and systole of mice. It can be seen that the thickness of the left ventricular posterior wall of the mice in the alcohol group (AF) was significantly increased compared with the control group (PF), and there was no significant difference in the thickness of the left ventricular posterior wall of the mice in the AF+ZJUIDS13 (heat-killed) group and the control group (PF). Therefore, it is judged that the inactivated ZJUIDS13 can effectively reduce the increase in left ventricular wall thickness caused by alcohol in mice and avoid the symptoms of myocardial hypertrophy caused by long-term alcohol intake.

[0095] like Fig.14 As shown, it is the heart weight and heart-to-body ratio of mice. It can be seen that the heart weight of mice in the alcohol group (AF) increased, and the heart-to-body ratio was higher. The heart weight and heart-to-body ratio of the AF+ZJUIDS13 (heat-killed) group were significantly reduced, and the symptoms of myocardial hypertrophy were restored.

[0096] like Fig.15 and Fig.16 As shown, it is the level of Bcl2 mRNA and Bax mRNA in mouse cardiomyocytes. It can be seen that the Bcl2 mRNA in the alcohol group (AF) group was significantly lower than that in the control group (PF), and the Bcl2 mRNA level in the AF+ZJUIDS13 (heat-killed) group was significantly restored compared with the alcohol group (AF). In addition, the Bax mRNA in the alcohol group (AF) group was significantly increased compared with the control group (PF), and the Bax mRNA level in the AF+ZJUIDS13 (heat-killed) group was significantly reduced compared with the alcohol group (AF). By analyzing the effects of long-term drinking on the levels of Bcl2 mRNA and Bax mRNA, it is shown that ZJUIDS13 can effectively reduce myocardial damage.

[0097] Finally, it should be noted that the above examples are only some specific embodiments of the present invention. Obviously, the present invention is not limited to the above embodiments, and there are many variations. All variations that can be directly derived or associated with the content disclosed by a person skilled in the art should be considered as the protection scope of the present invention.

Claims

1. Application of Pediococcus acidilactici ZJUIDS13 in the preparation of drugs for improving alcoholic cardiomyopathy.

2. The use according to claim 1, characterized in that: Protects against alcohol-induced myocardial damage.

3. The use according to claim 1 or 2, characterized in that At least one of the following: Reduce the level of creatine kinase isoenzyme (CK-MB) in serum, Restore myocardial capacity; Avoid alcohol-induced myocardial hypertrophy. Alleviate myocardial injury, including increasing Bcl2 mRNA and decreasing Bax mRNA.

4. The use according to any one of claims 1 to 3, characterized in that: Including Pediococcus acidilactici ZJUIDS13 and inactivated Pediococcus acidilactici ZJUIDS13.

5. The use according to claim 4, characterized in that: It includes a suspension prepared by using the bacterial powder of Pediococcus acidilactici ZJUIDS13 and an inactivated ZJUIDS13 bacterial liquid.

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