Application of post-exercise groin white fat-derived extracellular vesicles in preparation of medicine for preventing and / or treating myocardial ischemia-reperfusion injury

Abstract

The invention relates to application of extracellular vesicles derived from post-exercise groin white fat in preparation of a medicine for preventing and / or treating myocardial ischemia-reperfusion injury. The extracellular vesicles are derived from autokinetic white adipose tissues of groins, after long-term aerobic exercise (such as swimming or running), the extracellular vesicles in the white adipose tissues of groins are extracted, and it is found that the extracellular vesicles can remarkably reduce the myocardial infarction area caused by acute myocardial ischemia reperfusion, and the myocardial infarction effect is remarkably improved. And cardiac function decline and cardiac fibrosis caused by long-term myocardial ischemia reperfusion are inhibited. Experiments show that the exercised extracellular vesicles can inhibit apoptosis of myocardial cells. The invention provides a new treatment strategy for prevention and treatment of myocardial ischemia reperfusion injury.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the use of extracellular vesicles derived from post-exercise inguinal white fat in the preparation of a drug for preventing and / or treating myocardial ischemia-reperfusion injury. Background Art

[0002] Acute myocardial infarction is an extremely common cardiovascular disease with a high mortality rate. In clinical practice, the common treatment for acute myocardial infarction is to promptly reperfuse the ischemic area. However, when myocardial cells are in an ischemic state for a certain period of time, blood supply is restored. This process will damage myocardial cells, leading to adverse ventricular remodeling and decreased cardiac function. This damage is called myocardial ischemia / reperfusion (I / R) injury. Until now, there is still a lack of effective drugs for the prevention and treatment of myocardial ischemia / reperfusion injury. Summary of the invention

[0003] The purpose of the present invention is to provide the use of extracellular vesicles derived from post-exercise inguinal white fat in the preparation of a drug for preventing and / or treating myocardial ischemia-reperfusion injury. The extracellular vesicles of the present invention can effectively prevent and treat myocardial ischemia / reperfusion injury.

[0004] The present invention provides the use of extracellular vesicles derived from inguinal white fat after exercise in preparing a drug for preventing and / or treating myocardial ischemia-reperfusion injury.

[0005] Preferably, the exercise includes long-term regular aerobic exercise;

[0006] The long-term regularity refers to more than 4 consecutive weeks; the aerobic exercise includes swimming and / or running.

[0007] Preferably, the frequency of swimming and / or running is 1 to 2 times a day, each time for 10 to 90 minutes.

[0008] Preferably, the method for obtaining extracellular vesicles comprises the following steps:

[0009] The post-exercise inguinal white adipose tissue is digested and the extracellular vesicles are obtained by ultracentrifugation.

[0010] Preferably, the medicament comprises a liquid formulation.

[0011] Preferably, the dosage of the extracellular vesicles in the liquid preparation is 40 μg / 25 μL.

[0012] Preferably, obtaining the liquid preparation comprises dissolving the extracellular vesicles in PBS and / or physiological saline.

[0013] Preferably, the myocardial ischemia-reperfusion injury includes acute myocardial ischemia-reperfusion injury and / or chronic myocardial ischemia-reperfusion injury.

[0014] Preferably, the myocardial ischemia-reperfusion injury includes one or more of myocardial cell apoptosis, increase in myocardial infarction area, decreased cardiac function and cardiac fibrosis.

[0015] Preferably, the myocardial ischemia-reperfusion injury includes myocardial ischemia-reperfusion injury induced by oxygen-glucose deprivation and recovery.

[0016] The present invention provides the use of extracellular vesicles derived from inguinal white fat after exercise in the preparation of drugs for preventing and / or treating myocardial ischemia-reperfusion injury. The extracellular vesicles described in the present invention are derived from inguinal white adipose tissue after long-term exercise, and can effectively prevent and treat myocardial ischemia / reperfusion injury, including inhibiting myocardial cell apoptosis, reducing myocardial infarction area, inhibiting cardiac function decline and inhibiting cardiac fibrosis. Not only does it protect myocardial tissue at the cellular level and reduce the death of myocardial cells, it can also significantly reduce the area of ​​myocardial infarction caused by acute myocardial ischemia-reperfusion, effectively reduce the degree of damage to the myocardium caused by acute myocardial ischemia-reperfusion, and also have a significant inhibitory effect on the changes in cardiac structure and function caused by long-term myocardial ischemia-reperfusion injury, which helps to maintain the normal function of the heart and reduce the risk of progression of heart disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0018] Figure 1 The characteristic correlation diagram of the inguinal white fat-derived extracellular vesicles provided by the present invention; wherein A is a particle size analysis diagram, B is a transmission electron microscopy morphology diagram, and C is a protein immunoblot diagram;

[0019] Figure 2 This is an analysis diagram of the anti-cardiomyocyte apoptosis effect of mouse inguinal white fat-derived extracellular vesicles (Swim-iWAT-EVs) provided by the present invention; wherein A is a Tunel staining analysis diagram, B is a cell apoptosis statistical diagram, and C is a specific value of each group corresponding to Figure B; n=6, "*" represents p<0.05, and "**" represents p<0.01;

[0020] Figure 3This is an analysis diagram of the anti-cardiomyocyte apoptosis effect of epididymal white fat-derived extracellular vesicles provided by the present invention; wherein A is a Tunel staining analysis diagram, B is a cell apoptosis statistical diagram, and C is a specific value of each group corresponding to Figure B; n=6, "**" represents p<0.01, and "ns" represents no significant difference;

[0021] Figure 4 This is an analysis diagram of the anti-cardiomyocyte apoptosis effect of extracellular vesicles derived from inguinal white fat of running mice (Run-iWAT-EVs) provided by the present invention; wherein A is a Tunel staining analysis diagram, B is a cell apoptosis statistical diagram, and C is the specific value of each group corresponding to Figure B; n=6, "*" represents p<0.05, and "**" represents p<0.01;

[0022] Figure 5 The myocardial infarction area analysis diagram 24 hours after myocardial I / R surgery provided by the present invention; wherein A is a representative diagram of TTC staining, B is a statistical diagram of ischemic area, C is a statistical diagram of infarction area, and D is a specific value of each group corresponding to diagrams B and C; n=7-10, "*" represents p<0.05, and "**" represents p<0.01;

[0023] Figure 6 The cardiac function and cardiac fibrosis analysis diagrams 2 weeks after myocardial I / R surgery provided by the present invention; wherein A is an echocardiogram, B is an ejection fraction (EF) statistical diagram, C is a short-axis shortening rate (FS) statistical diagram, D is a Masson staining analysis diagram, E is a cardiac fibrosis statistical diagram, and F is a specific numerical value corresponding to each group of B, C and E diagrams; n=7-9, "*" represents p<0.05, and "**" represents p<0.01. DETAILED DESCRIPTION

[0024] The present invention provides the use of extracellular vesicles derived from inguinal white fat after exercise in the preparation of drugs for preventing and / or treating myocardial ischemia-reperfusion injury. In a specific embodiment, the extracellular vesicles derived from inguinal white fat after exercise can effectively prevent and treat myocardial ischemia / reperfusion injury, including inhibiting myocardial cell apoptosis, reducing myocardial infarction area, inhibiting cardiac function decline and inhibiting cardiac fibrosis. In a specific embodiment, the extracellular vesicles derived from inguinal white fat after exercise not only protect myocardial tissue at the cellular level and reduce the death of myocardial cells, but also can significantly reduce the myocardial infarction area caused by acute myocardial ischemia-reperfusion, and effectively reduce the degree of damage to the myocardium caused by acute myocardial ischemia-reperfusion. In a specific embodiment, the extracellular vesicles derived from inguinal white fat after exercise also have a significant inhibitory effect on the changes in cardiac structure and function caused by long-term myocardial ischemia-reperfusion injury, which helps to maintain the normal function of the heart and reduce the risk of progression of heart disease.

[0025] In a specific embodiment, the exercise includes long-term regular aerobic exercise; the long-term regularity refers to more than 4 consecutive weeks; the aerobic exercise includes swimming and / or running. In a specific embodiment, the long-term regular aerobic exercise can also be aerobic exercise for more than 12 consecutive weeks. In a specific embodiment, the frequency of swimming and / or running can be 1 to 2 times a day, and each time can last for 10 to 90 minutes. In a specific embodiment, when the exercise is running, the running speed includes 5m / min to 15m / min.

[0026] In a specific embodiment, the swimming can be gradually increased from 10 minutes per day, and further, it can be started from 10 minutes per day, and the swimming time can be increased by 10 minutes each day based on the previous day, and gradually increased to 90 minutes per day. In a specific embodiment, the running speed can be started from 5m / min and gradually increased to 15m / min, and further, it can be started from running at a speed of 5m / min for 10 minutes, and the speed can be increased by 2m / min and the time can be increased by 10 minutes every day based on the previous day, and gradually increased to running at a speed of 15m / min for 1 hour every day.

[0027] In a specific embodiment, the method for obtaining extracellular vesicles may include the following steps:

[0028] The post-exercise inguinal white adipose tissue is digested and the extracellular vesicles are obtained by ultracentrifugation.

[0029] The present invention performs digestion treatment on the post-exercise inguinal white adipose tissue. In a specific embodiment, the digestion treatment may be to treat the post-exercise inguinal white adipose tissue with a digestive enzyme. In a specific embodiment, the digestive enzyme may be collagenase, and may further be a collagenase solution with a mass ratio of 1%. In a specific embodiment, the digestion treatment may be to add collagenase and then digest for 40 minutes at 37°C using a shaker. In a specific embodiment, the digestion treatment may terminate the digestion by adding an equal volume of 2μM EGTA.

[0030] After digestion of the post-exercise inguinal white adipose tissue, the present invention obtains the extracellular vesicles by ultracentrifugation. In a specific embodiment, the ultracentrifugation can be performed multiple times at 4°C, more specifically: 300g centrifugation for 10 minutes, taking the supernatant and repeating the centrifugation once; 2000g centrifugation for 20 minutes, taking the supernatant and repeating the centrifugation once; 12000g centrifugation for 30 minutes, taking the supernatant and repeating the centrifugation once; finally 100000g centrifugation for 70 minutes, and collecting the precipitate (i.e., the post-exercise inguinal white adipose tissue extracellular vesicles). In a specific embodiment, after obtaining the precipitate, PBS can be added to resuspend the precipitate to make an extracellular vesicle suspension for subsequent experiments.

[0031] In certain embodiments, the drug may include a liquid formulation.

[0032] In a specific embodiment, the dosage of the extracellular vesicles in the liquid preparation is 40 μg / 25 μL.

[0033] In a specific embodiment, obtaining the liquid preparation comprises dissolving the extracellular vesicles in PBS and / or physiological saline.

[0034] In a specific embodiment, the myocardial ischemia-reperfusion injury includes acute myocardial ischemia-reperfusion injury and / or chronic myocardial ischemia-reperfusion injury.

[0035] In a specific embodiment, the myocardial ischemia-reperfusion injury includes one or more of myocardial cell apoptosis, increase in myocardial infarction area, decreased cardiac function and cardiac fibrosis.

[0036] In a specific embodiment, the myocardial ischemia-reperfusion injury includes myocardial ischemia-reperfusion injury induced by oxygen-glucose deprivation and recovery.

[0037] To further illustrate the present invention, the application of the extracellular vesicles derived from post-exercise inguinal white fat provided by the present invention in the preparation of drugs for preventing and / or treating myocardial ischemia-reperfusion injury is described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0038] Example 1

[0039] Characterization of extracellular vesicles derived from inguinal white fat in swimming and sedentary mice.

[0040] 1 Construction of swimming exercise mouse model.

[0041] Eight-week-old male C57BL / 6 mice were subjected to 4 weeks of swimming exercise. They started with 10 minutes of swimming exercise each time, and then the swimming time was increased by 10 minutes each day based on the previous day until each swimming time reached 90 minutes. They swam twice a day, with an interval of 4 to 6 hours in between. Mice of the same age were sedentary as controls. After 28 days, the inguinal white fat of the swimming and sedentary mice was taken for extraction of extracellular vesicles.

[0042] 2 Extraction of extracellular vesicles from inguinal white fat.

[0043] Cut the inguinal white fat into small pieces, add 1% collagenase, and digest on a shaker at 37°C for 40 minutes. Then add an equal volume of 2μM EGTA to terminate the digestion. Centrifuge several times at 4°C, first centrifuge at 300g for 10 minutes, aspirate the supernatant into a new centrifuge tube, repeat the centrifugation once; centrifuge at 2000g for 20 minutes, aspirate the supernatant into a new centrifuge tube; centrifuge at 12000g for 30 minutes, aspirate the supernatant into a new centrifuge tube; centrifuge at 100000g for 70 minutes. The resulting precipitate is the extracellular vesicles. Discard the supernatant, resuspend in PBS, and use for subsequent experiments.

[0044] 3 Characterization and identification of extracellular vesicles.

[0045] Take 1 μl of the isolated and purified extracellular vesicles, dilute them 1000 to 25000 times with PBS, and use ZetaView instrument to analyze the particle size distribution. The results are as follows: Figure 1 As shown in A, the diameter of the obtained extracellular vesicles is about 100 nm.

[0046] Take 20 μl of separated and purified extracellular vesicles and drop them on the copper mesh. Then add 20 μl of phosphotungstic acid dye solution on the copper mesh and negatively stain for 1 minute. Then use a transmission electron microscope to observe the structure of the extracellular vesicles. The results are as follows: Figure 1 As shown in B, the obtained extracellular vesicles are spherical with a depression in the middle and have typical morphological characteristics of extracellular vesicles.

[0047] Take the isolated and purified extracellular vesicles, add protein lysis buffer to fully lyse them, then centrifuge at 12000rpm, 4℃ for 30min, add loading buffer to the obtained cell supernatant, and place it in a metal bath at 100℃ for 10min. The obtained sample was analyzed by protein immunoblotting for surface marker proteins of extracellular vesicles. The results are as follows Figure 1 As shown in C, the obtained extracellular vesicles expressed extracellular vesicle-specific proteins CD63, CD9, Alix and Tsg101, but did not express the negative control protein Calnexin.

[0048] Example 2

[0049] Swim-iWAT-EVs can significantly inhibit OGD / R-induced cardiomyocyte apoptosis.

[0050] 1Isolation and culture of neonatal rat primary cardiomyocytes (NRCM).

[0051] Take the heart of a newborn SD rat in the clean bench and rinse it in ADS buffer. Cut the heart into a homogenate, then transfer it to a sterilized glass bottle, add digestion solution (0.06% trypsin + 0.04% collagenase mixture), blow it evenly, and place it on a 37°C shaker for digestion for 20 minutes. Then add horse serum to stop digestion, centrifuge it at 1000rpm for 5 minutes, discard the supernatant, and add a culture medium containing 10% fetal bovine serum to resuspend the precipitate. Continue to add digestion solution and blow it several times, place it on a shaker for digestion, and digest it repeatedly until the tissue is completely digested. Add cardiomyocyte culture medium (DMEM + 1% penicillin and streptomycin + 10% fetal bovine serum + 5% horse serum) to the cell precipitate. Filter it with a 100μm pore size cell strainer, add the filtered cell suspension to the cell culture dish, and place it in a cell culture incubator for adherent culture. After one hour, collect the non-adherent cell suspension, centrifuge it at 1000rpm at room temperature for 5 minutes, and discard the supernatant. Resuspend the cell precipitate with ADS buffer and centrifuge it at 3000rpm Percoll density gradient for 30 minutes. The middle layer is the NRCM cardiomyocytes. The middle layer cells were resuspended in cardiomyocyte culture medium and used for subsequent experiments after 24 hours of attachment.

[0052] 2. Construction of oxygen glucose deprivation / recovery (OGD / R) model and extracellular vesicle treatment.

[0053] After the NRCM cells adhered to the wall, the culture medium was replaced with a culture medium that removed extracellular vesicles (fetal bovine serum and horse serum were removed by ultracentrifugation (4°C, 100,000g centrifugation for 90 minutes)). At the same time, extracellular vesicles (40 μg / ml) from swimming and sitting inguinal white fat were co-cultured for 40 hours, and then replaced with sugar-free culture medium, and the above extracellular vesicles were added again. The cell culture plate was placed in a negative oxygen box and incubated in a constant temperature box for 8 hours. After that, the cells were cultured with resugar and reoxygenation, and the cells were harvested after 12 hours for Tunel staining.

[0054] 3Tunel staining was used to detect cardiomyocyte apoptosis.

[0055] The apoptosis of NRCM cells was detected by Tunel staining kit (Novozyme, A112). The staining results were as follows: Figure 2 A in Figure 2 B and Figure 2As shown in Figure C, compared with the control group, the proportion of Tunel-positive cells in the OGD / R group increased significantly, indicating a significant increase in cell apoptosis. Swim-iWAT-EVs can significantly reduce the increase in Tunel-positive cells caused by OGD / R, indicating that Swim-iWAT-EVs can inhibit OGD / R-induced cardiomyocyte apoptosis, while Sed-iWAT-EVs do not have this effect. These results indicate that inguinal white fat-derived extracellular vesicles themselves do not have a cardioprotective effect, but long-term aerobic exercise causes extracellular vesicles from this source to produce a cardioprotective effect.

[0056] Example 3

[0057] Epididymal white adipose-derived extracellular vesicles cannot protect against OGD / R-induced cardiomyocyte apoptosis.

[0058] 1 Extraction of extracellular vesicles from epididymal white fat.

[0059] The epididymal white fat from the swimming and sitting mice in Example 1 was respectively used to extract extracellular vesicles. The extraction of the extracellular vesicles was the same as the extraction of extracellular vesicles from the inguinal white fat in Example 1.

[0060] 2 Effect of epididymal white fat-derived extracellular vesicles on cardiomyocyte apoptosis.

[0061] NRCM cells were treated with extracellular vesicles derived from epididymal white fat of swimming and sitting mice (Swim-eWAT-EVs and Sed-iWAT-EVs, respectively), and the rest of the operations (NRCM isolation and culture, OGD / R model construction and extracellular vesicle treatment, Tunel staining) were the same as in Example 2. The Tunel staining results are shown in Figure 3 A in Figure 3 B and Figure 3 As shown in Figure C, compared with the control group, the apoptosis of cells in the OGD / R group increased significantly, but Swim-eWAT-EVs and Sed-iWAT-EVs had no inhibitory effect on OGD / R-induced cardiomyocyte apoptosis. This result reflects the difference in the function of adipose tissue in different parts. Compared with epididymal white fat, the extracellular vesicles secreted by inguinal white fat after aerobic exercise have a cardioprotective effect.

[0062] Example 4

[0063] Run-iWAT-EVs can significantly inhibit OGD / R-induced cardiomyocyte apoptosis.

[0064] 1 Construction of running exercise mouse model.

[0065] Eight-week-old male C57BL / 6 mice were subjected to 12 weeks of running exercise. On the first day, the mice were placed on a treadmill to adapt for 10-15 minutes, then ran at a speed of 5m / min for 10 minutes, and then the speed increased by 2m / min and the time increased by 10min every day until the speed was 15m / min and the time was 1h on the 6th day. After that, they ran at a speed of 15m / min for 1h every day for 12 weeks. Mice of the same age were sedentary as controls.

[0066] 2 Extraction of extracellular vesicles from inguinal white fat of running mice.

[0067] After the above-mentioned running exercise, the inguinal white fat of the running and sitting mice was respectively taken to extract extracellular vesicles. The extraction of the extracellular vesicles was the same as the extraction of extracellular vesicles from inguinal white fat in Example 1.

[0068] 3 Effect of extracellular vesicles derived from inguinal white fat of running and sitting mice on cardiomyocyte apoptosis.

[0069] NRCM cells were treated with extracellular vesicles from inguinal white fat of running and sitting mice, respectively, and the rest of the operations (NRCM isolation and culture, OGD / R model construction and extracellular vesicle treatment, Tunel staining) were the same as in Example 2. The Tunel staining results are shown in Figure 4 A in Figure 4 B and Figure 4 As shown in Figure C, compared with the control group, cell apoptosis in the OGD / R group increased significantly, and Run-iWAT-EVs could significantly inhibit OGD / R-induced cardiomyocyte apoptosis, while Sed-iWAT-EVs did not have this effect. This result further indicates that inguinal white fat-derived extracellular vesicles themselves do not have a cardioprotective effect, but long-term aerobic exercise causes extracellular vesicles from this source to produce a cardioprotective effect.

[0070] Example 5

[0071] Swim-iWAT-EVs can reduce myocardial infarction area caused by acute myocardial I / R.

[0072] 1 Construction of acute myocardial I / R injury model in mice and treatment with extracellular vesicles.

[0073] The mouse chest was depilated. After the mouse was anesthetized, it was fixed on a 37℃ constant temperature pad with medical tape. A small incision was made in the neck to expose the trachea and intubate the trachea. Use a ventilator to assist breathing, maintain a respiratory rate of 120 times / min, and a tidal volume of 2mL. A straight incision of about 1cm was made about 0.5cm below the armpit, and the skin and pectoralis major muscle were cut layer by layer to expose the intercostal muscles. Blunt elbow ophthalmic forceps were inserted between the third and fourth ribs, and the intercostal muscles were bluntly separated to expose the heart. A low-elastic 7-0 suture was used to tie a slipknot at about 1 / 3 of the line connecting the left atrial appendage to the apex of the heart. The whitening below the ligature to the apex of the heart indicated that the left anterior descending branch of the coronary artery was successfully ligated. Clamp the wound skin with hemostatic forceps and temporarily close the chest cavity. After 30 minutes, the chest cavity was opened again and the ligature was untied. Suture the ribs, pectoralis major muscle and skin in sequence with 4-0 sutures. When the tail clamping reflex of the mouse was partially recovered, the endotracheal tube was removed, and the tracheal secretions were sucked away with sterile gauze, but the tracheal wound was not sutured. The tape that fixed the mouse was torn off, and the mouse was placed on a 37°C constant temperature pad and returned to the cage after it woke up. The sham operation group did not ligate the left anterior descending coronary artery, and the rest of the treatment was the same as the surgery group. After ligating the left anterior descending coronary artery, extracellular vesicles were injected into the ischemic part of the heart through the myocardium at multiple points, and each mouse was given 40μg of extracellular vesicles (dissolved in 25μL PBS). The experiment was terminated 24 hours after surgery. There were 7 mice in the sham operation group, and 10 mice in the Swim-iWAT-EVs treatment group and the Sed-iWAT-EVs treatment group.

[0074] 2TTC staining.

[0075] 24 hours after myocardial I / R surgery, the mice were anesthetized, the left anterior descending branch of the coronary artery was ligated again, and 1 mL of 1% Evans blue dye was injected into the left ventricle. The heart was then removed and myocardial infarction was detected by TTC staining. The samples were observed and photographed under a stereomicroscope. Image J software was used to count the overall area of ​​the front and back of the slices, the white infarct area, and the red ischemic area. The ischemic area / left ventricular area (AAR / LV) and the infarct area / ischemic area (INF / AAR) were calculated. The results are shown in the figure. Figure 5 A in Figure 5 B in Figure 5 C and Figure 5 As shown in D, compared with PBS treatment, Swim-iWAT-EVs can significantly reduce the myocardial infarction area caused by I / R, while Sed-iWAT-EVs do not have this effect. These results indicate that Swim-iWAT-EVs can significantly reduce acute myocardial I / R injury in vivo, while Sed-iWAT-EVs do not have this effect.

[0076] Example 6

[0077] Swim-iWAT-EVs can inhibit the decline in cardiac function and cardiac fibrosis caused by long-term myocardial I / R.

[0078] 1 Construction of long-term myocardial I / R injury model in mice and treatment with extracellular vesicles.

[0079] Myocardial I / R surgery and extracellular vesicle treatment were consistent with the aforementioned acute myocardial I / R injury model. The experiment was terminated 2 weeks after surgery for subsequent detection and analysis. There were 9 mice in the I / R 2w group, 7 mice in the sham operation group, the I / R 2w+Swim-iWAT-EVs group, and the I / R 2w+Sed-iWAT-EVs group.

[0080] 2. Echocardiography to detect heart function.

[0081] Two weeks after myocardial I / R surgery (I / R2w), the mice were examined by cardiac echocardiography using the VisualSonics Vevo 2100 system to evaluate the improvement of their cardiac function. Figure 6 As shown in A to C, sham represents the sham operation group with normal cardiac function. Compared with the sham group, the cardiac function of I / R 2w mice was significantly deteriorated. Swim-iWAT-EVs can significantly inhibit the decline of cardiac function in I / R2w mice, while Sed-iWAT-EVs do not have this effect.

[0082] 3 Masson staining was used to detect cardiac fibrosis.

[0083] Two weeks after myocardial I / R surgery, heart tissue was obtained and made into paraffin sections, and the fibrosis of heart tissue was detected by Masson staining. Figure 6 As shown in D to F in Figure 1, myocardial tissue cells are red, cell nuclei are blue-black, and collagen fibers are blue. Compared with the sham group, cardiac fibrosis in I / R 2w mice increased significantly, indicating that Swim-iWAT-EVs can significantly inhibit cardiac fibrosis in I / R 2w mice, while Sed-iWAT-EVs do not have this effect.

[0084] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creative work, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. Application of extracellular vesicles derived from inguinal white fat after exercise in the preparation of drugs for preventing and / or treating myocardial ischemia-reperfusion injury.

2. The use according to claim 1, characterized in that: The exercise includes long-term regular aerobic exercise; The long-term pattern is defined as more than 4 consecutive weeks; The aerobic exercise includes swimming and / or running.

3. The use according to claim 2, characterized in that: The frequency of swimming and / or running is 1 to 2 times a day, each time for 10 to 90 minutes.

4. The use according to claim 1, characterized in that: The method for obtaining extracellular vesicles comprises the following steps: The post-exercise inguinal white adipose tissue is digested and the extracellular vesicles are obtained by ultracentrifugation.

5. The use according to claim 1, characterized in that: The medicament includes a liquid preparation.

6. The use according to claim 5, characterized in that: The dosage of the extracellular vesicles in the liquid preparation is 40 μg / 25 μL.

7. The use according to claim 5, characterized in that: The obtaining of the liquid preparation comprises dissolving the extracellular vesicles in PBS and / or physiological saline.

8. The use according to claim 1, characterized in that: The myocardial ischemia-reperfusion injury includes acute myocardial ischemia-reperfusion injury and / or long-term myocardial ischemia-reperfusion injury.

9. The use according to claim 1, characterized in that: The myocardial ischemia-reperfusion injury includes one or more of the following: myocardial cell apoptosis, increase in myocardial infarction area, decreased cardiac function and cardiac fibrosis.

10. The use according to claim 1, characterized in that: The myocardial ischemia-reperfusion injury includes myocardial ischemia-reperfusion injury induced by oxygen-glucose deprivation and recovery.

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