He-tian summer apple beverage, and preparation method and application thereof

Abstract

This invention discloses a Hotan summer apple beverage, its preparation method, and its application, belonging to the field of pharmaceutical technology. The Hotan summer apple beverage of this invention comprises the following components, by weight: 500-1000 parts Hotan summer apple extract, 100-200 parts Hotan red grape extract, 1-2 parts Hotan rose, 0.1-0.5 parts saffron, and 20-60 parts flavoring agent. This Hotan summer apple beverage of this invention can significantly increase the proliferation level of H9C2 cells damaged by oxygen-glucose deprivation, and significantly decrease the expression of the serum myocardial injury marker lactate dehydrogenase (LDH), indicating that the beverage of this invention has a protective effect on cardiomyocytes. Compared with the commonly used inhibitor Acetylcysteine, this Hotan summer apple beverage of this invention is made from ingredients that are both food and medicine, with no side effects from long-term use, making it more suitable for long-term use by patients with chronic diseases. The materials in this beverage are Hotan summer apple, Hotan red grape, Hotan rose, and saffron, which are products of large-scale cultivation, with high yield and low cost. The preparation process is simple, reducing the cost of medication.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a Hotan summer apple beverage, its preparation method and application, and in particular, the application of the Hotan summer apple beverage in the preparation of drugs for treating myocardial injury. Background Technology

[0002] Myocardial injury refers to the phenomenon of damage or necrosis of myocardial cells. Diseases caused by myocardial injury include myocarditis, coronary atherosclerotic heart disease, myocardial ischemia, and acute myocardial infarction, which seriously endanger life.

[0003] Current treatments for myocardial injury primarily involve medication, interventional therapy, surgery, and rehabilitation. However, interventional and surgical treatments can also cause some damage to myocardial cells. Current medication treatments, depending on the underlying cause, mainly utilize nitrates (such as nitroglycerin), beta-blockers, antiplatelet drugs, statins, and myocardial nutrients; or cardiotonics (such as digitalis) and phosphates. However, these medications have certain side effects. Summary of the Invention

[0004] To address the aforementioned issues, this invention proposes a Hotan summer apple beverage, its preparation method, and its application. This beverage can regulate myocardial cell proliferation and repair myocardial damage, possessing the properties of both food and medicine, and has minimal side effects on the human body.

[0005] To achieve the above objectives, the present invention is implemented through the following technical solution: The first objective of this invention is to provide a Hotan summer apple beverage, which comprises the following components in parts by weight: 500-1000 parts of Hotan summer apple extract, 100-200 parts of Hotan red grape extract, 1-2 parts of Hotan rose, 0.1-0.5 parts of saffron, and 20-60 parts of flavoring agent.

[0006] Furthermore, the flavoring agent is mainly a sweetener, selected from white sugar, honey, xylose, maltitol, and lactose.

[0007] The second objective of this invention is to provide a method for preparing a Hotan summer apple beverage, specifically including the following steps: (1) Add water to the Hetian rose and heat and reflux to extract 2-3 times, 1-2 hours each time. Filter and combine the filtrates to form rose extract for later use. (2) Take saffron, add water, heat and reflux to extract 2-3 times, 1-2 hours each time, filter, and combine the filtrates to form saffron extract for later use; (3) Combine the extracts of Hetian summer apple, Hetian red grape, rose extract and saffron extract, add flavoring agent and mix thoroughly. Let stand at 0-4℃ for 12 hours, filter, bottle and sterilize.

[0008] Furthermore, in step (1), the solid-liquid ratio in the extraction of Hetian rose petals by adding water and heating and refluxing is 1:8-12.

[0009] Furthermore, in step (2), the solid-liquid ratio in the saffron extraction by heating and reflux with water is 1:20-30.

[0010] Furthermore, the Hotan summer apple extract in step (3) is selected from fresh Hotan summer apple juice or Hotan summer apple dried water extract; the Hotan red grape extract is selected from fresh Hotan red grape juice or Hotan red grape dried water extract.

[0011] Furthermore, the water extract of Hetian summer apple described in step (3) is obtained by adding water to Hetian summer apple and heating and refluxing 2-3 times, each time for 1-2 hours, filtering, and combining the filtrates. The water extract of Hetian red raisins described in step (3) is obtained by adding water to Hetian red raisins and heating and refluxing 2-3 times, each time for 1-2 hours, filtering, and combining the filtrates.

[0012] Furthermore, in the extraction of dried Hetian summer apples by heating and reflux with water, the solid-liquid ratio is 1:8-12.

[0013] Furthermore, in the extraction of Hetian red raisins by adding water and heating and refluxing, the solid-liquid ratio is 1:8-12.

[0014] Furthermore, the heating temperature in the heating reflux in step (1) is 90-100℃; the heating temperature in the heating reflux in step (2) is 90-100℃; the heating temperature in the heating reflux in step (3) is 90-100℃.

[0015] The third objective of this invention is to provide the application of Hotan summer apple beverage in the preparation of drugs for the prevention or treatment of myocardial injury.

[0016] Furthermore, the drug is an oral dosage form.

[0017] Furthermore, the drug also contains pharmaceutically acceptable excipients.

[0018] The beneficial effects of this invention, including the Hotan summer apple beverage, its preparation method, and its application, are as follows: (1) The summer apple beverage of the present invention can significantly increase the proliferation level of H9C2 cells with myocardial injury caused by oxygen and sugar deprivation, and significantly reduce the expression of serum myocardial injury marker lactate dehydrogenase (LDH), indicating that the beverage of the present invention has a protective effect on myocardial cells.

[0019] (2) Compared with the commonly used inhibitor Acetylcysteine, the Hetian Xia apple drink of the present invention is a food and medicine homology ingredient, which has no side effects when used for a long time and is more suitable for long-term use by patients with chronic diseases.

[0020] (3) The ingredients in the beverage of the present invention are Hotan summer apples, Hotan red grapes, Hotan roses and saffron, which are products grown on a large scale, with high yield and low cost. The preparation process is simple and reduces the cost of medication. Attached Figure Description

[0021] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0022] Figure 1 This is a bar chart of the effect of Hetian summer apple beverage (therapeutic drug A) on H9C2 cell proliferation in Example 1 of the present invention; Figure 2 This is a bar chart showing the effect of Hetian summer apple beverage (therapeutic drug B) on H9C2 cell proliferation in Example 2 of the present invention; Figure 3 This is a scatter plot of H9C2 cell survival rates in the control and model groups under an oxygen-glucose deprivation injury model. Figure 4 This is a bar chart showing the LDH activity in the supernatant of H9C2 cells in the control and model groups under an oxygen-glucose deprivation injury model. Figure 5 This is a scatter plot of cell survival rates in each group under the oxygen-glucose deprivation injury model. Figure 6 This is a bar chart of LDH activity in the cell supernatant of each group under the oxygen-glucose deprivation injury model. Detailed Implementation

[0023] The present invention will be further described in detail below with reference to specific embodiments and accompanying drawings.

[0024] Example 1 A method for preparing a Hotan summer apple beverage includes the following steps: (1) Take 20g of Hetian rose (Damask rose) petals, add 200ml of water and heat under reflux for 1 hour, filter, and set aside the filtrate. Add 100ml of water to the residue and heat under reflux again at 90-100℃ for 1 hour, filter, and combine the filtrate with the first filtrate to form rose extract, set aside. (2) Take 2g of saffron, heat and reflux twice with 50ml of water at 90-100℃, filter, and combine the filtrates to form saffron extract for later use; (3) Take 8-10 kg of fresh Hotan summer apples and juice them to obtain 6000 mL of fresh apple juice; take 1.5-2 kg of fresh Hotan red grapes and juice them to obtain 1200 mL of fresh red grape juice; mix the fresh apple juice, fresh red grape juice, rose extract obtained in step (1), and saffron extract obtained in step (2), add 500 g of white sugar as a flavoring agent and dissolve them completely, make up to 10000 ml with water, let stand at 0-4℃ for 12 hours, and filter. Bottle in pressure-resistant and heat-resistant glass bottles and sterilize at 121℃ for 15 minutes to obtain Hotan summer apple beverage.

[0025] Example 2 A method for preparing a Hotan summer apple beverage includes the following steps: (1) Take 20g of Hetian rose (Damask rose) petals, add 200ml of water and heat at 90-100℃ under reflux for 1 hour, filter, and set aside the filtrate. Add 100ml of water to the residue and heat under reflux again for 1 hour, filter, and combine the filtrate with the first filtrate to form rose extract, set aside. (2) Take 2g of saffron, heat and reflux twice with 50ml of water at 90-100℃, filter, and combine the filtrates to form saffron extract for later use; (3) Take 1 kg of dried summer apples from Hotan, add water at a solid-liquid ratio of 1:9, heat and reflux at 90-100℃ twice for one hour each time, filter, and combine the filtrates to obtain the water extract of dried summer apples from Hotan for later use. Take 200g of Hetian red raisins, add water at a solid-liquid ratio of 1:9, heat and reflux at 90-100℃ twice for one hour each time, filter, and combine the filtrates to obtain the water extract of Hetian red raisins for later use. Mix the dried water extracts of Hetian summer apples and Hetian red raisins with the rose extract obtained in step (1) and the saffron extract obtained in step (2), then add 500g of white sugar as a flavoring agent and dissolve completely. Make up the volume to 10000ml with water, let stand at 0-4℃ for 12 hours, and filter. Bottle in pressure-resistant and heat-resistant glass bottles and sterilize at 121℃ for 15 minutes to obtain the Hetian summer apple beverage.

[0026] Example 3 This invention verifies the effects and mechanisms of Hetian summer apple beverage on cardiomyocyte proliferation and apoptosis under oxygen-glucose deprivation injury. I. Experimental Materials 1.1 Experimental Cells H9C2 rat cardiomyocytes were derived from Pronosei Biotechnology. The cell culture conditions were DMEM (high glucose) medium + 10% FBS + 1% PS, 37℃, 5% CO2, and saturated humidity.

[0027] 1.1 Experimental Reagents and Consumables Table 1 Experimental Reagents

[0028] 1.2 Experimental Apparatus CO2 cell incubator, model Smart Cell HF-90; biosafety cabinet, model HF1200LC; benchtop low-speed centrifuge, model DK-80; constant temperature water bath, model TGL-16GB; -20℃ freezer, model BCD-249LCK; -80℃ freezer, model DW-HL388; manual single-channel pipette, model Research plus; liquid nitrogen tank, model YDS-50-125; fluorescence inverted microscope, model Eclipse TS100-F; benchtop centrifuge, model Neofuge; pipette, model Dragon lab; microplate reader, model xMarkTM.

[0029] II. Experimental Methods 2.1 Basic Cell Culture Operations 2.1.1 Cell resuscitation H9C2 rat cardiomyocytes were taken from the liquid nitrogen and immediately thawed in a 37°C water bath. The thawed rat cardiomyocytes were then quickly added to a 15mL centrifuge tube containing 9mL of complete culture medium and centrifuged at 1000r / min for 5min. The supernatant was discarded, and the rat cardiomyocytes were seeded into culture flasks and cultured in a 37°C, saturated humidity, 5% CO2 cell culture incubator.

[0030] 2.1.2 Cell passage Discard the culture medium in the bottle, add 3 mL of sterile PBS buffer and wash repeatedly, discard the PBS buffer, add 1 mL of trypsin to the bottle, spread the trypsin solution evenly on the cell layer, gently shake the cell culture flask, and digest in an incubator. After 30 seconds, add 1 mL of culture medium containing 10% FBS to stop the digestion of trypsin, and repeatedly rinse the bottom of the culture flask with a pipette tip to wash off any undetached cells. Add the cell suspension to a 15 mL centrifuge tube, centrifuge at 1000 r / min for 5 min, carefully aspirate and discard the liquid in the centrifuge tube, and resuspend the cells with 2 mL of complete culture medium. Observe the cells under an inverted microscope, adjust the appropriate cell density, seed the cells into a culture flask, and place it in an incubator for culture.

[0031] 2.1.3 Cell cryopreservation Prepare cryovials in advance, label the cryovials with cell information, and add 400 μL FBS and 100 μL DMSO to the cryovials to dilute the cryopreservation solution and mix thoroughly; observe the culture flask (25 cm). 2 When rat cardiomyocytes adhered and fused to the wall for 90% of the time, the cells were digested with trypsin, centrifuged and the supernatant was discarded. The cells were resuspended in 500 μL of complete culture medium and added to cryovials and mixed by inversion. After the cells in the cryovials were cooled to 4℃ (20 min), -20℃ (30 min), and -80℃ (overnight), they were transferred to liquid nitrogen for cryopreservation.

[0032] 2.2 Screening of therapeutic drug intervention concentrations H9C2 cells with good growth and a confluence rate of 90% were collected and prepared into 5×10⁶ cells using complete culture medium. 4 Single-cell suspensions of cells / mL were seeded into 96-well plates (100 μL / well, 5 replicates). After 24 h of cell adhesion, cells were treated with different concentrations of treatment drug A (likely referring to the drug in Example 1) (0, 10, 20, 40, 80, 160 mg / mL) and treatment drug B (likely referring to the drug in Example 2) (0, 1, 5, 10, 20, 40 mg / mL) for 24 h. After treatment, the culture medium in each well was aspirated, and 100 μL of prepared 10% CCK-8 solution was added to each well. The OD value at 450 nm was measured using a microplate reader after 1 h. All data are expressed as mean ± standard deviation. The data were statistically analyzed using SPSS 19.0 software, employing a one-way ANOVA method. A p-value < 0.05 indicated a significant difference. GraphPad Prism 5.0 was used for plotting. The results are shown in Tables 2 and 3. Figure 1 and Figure 2 As shown.

[0033] Table 2. Effects of drug A intervention on H9C2 cell proliferation ( (n=5)

[0034] Note: △ Compared with treatment drug A (0 mg / ml), P<0.05; ▲ Compared with treatment drug A (10 mg / ml), P<0.05; ▽ Compared with treatment drug A (20 mg / ml), P<0.05; ▼ Compared with treatment drug A (40 mg / ml), P<0.05; ☆ Compared with treatment drug A (80 mg / ml), P<0.05.

[0035] Table 3. Effects of therapeutic drug B intervention on H9C2 cell proliferation levels ( (n=5)

[0036] Note: △ Compared with treatment drug B (0 mg / ml), P<0.05; ▲ Compared with treatment drug B (1 mg / ml), P<0.05; ▽ Compared with treatment drug B (5 mg / ml), P<0.05; ▼ Compared with treatment drug B (10 mg / ml), P<0.05; ☆ Compared with treatment drug B (20 mg / ml), P<0.05.

[0037] 2.3 Mechanism of action of therapeutic drugs in promoting cell proliferation 2.3.1 OGD / R Model Construction H9C2 cells were transferred to a hypoxic incubator at 37°C (5% CO2, 94% N2, 1% O2) and cultured in glucose-free DMEM. After 6 hours, the DMEM (high glucose) medium was replaced, and the cells were transferred to a conventional incubator and cultured for another 12 hours.

[0038] 2.3.2 Model Validation: Cell Proliferation Detection H9C2 cells with good growth and a confluence rate of 90% were collected and prepared into 5×10⁶ cells using complete culture medium. 4 Single-cell suspensions of cells / mL were seeded into 96-well plates (100 μL / well, 5 replicates). After 24 h of cell adhesion, an OGD / R model was constructed, and control cells were prepared simultaneously.

[0039] After the intervention, the culture medium in each well was aspirated, and 100 μL of prepared 10% CCK-8 solution was added to each well. The OD value at 450 nm was measured using a microplate reader 1 h later. All data are expressed as mean ± standard deviation. The data were statistically analyzed using SPSS 19.0 software, employing a one-way ANOVA method. A p-value < 0.05 indicated a significant difference. GraphPad Prism 5.0 was used for plotting, and the results are shown in Table 4. Figure 3 As shown: Table 4. Cell viability analysis of each group ( )

[0040] Note: △ Compared with the control group, P<0.05.

[0041] 2.3.2 Model Validation: LDH Content Detection H9C2 cells with good growth and a confluence rate of 90% were collected and prepared into 5×10⁶ cells using complete culture medium. 4Single-cell suspensions of cells / mL were seeded into 96-well plates (100 μL / well, 3 replicates). After 24 h of cell adhesion, an OGD / R model was constructed, establishing the model group cells. Simultaneously, blank and control groups were prepared. The model group cells were treated with Tianxia apple beverage (therapeutic drug A) as described in Example 1 and Tianxia apple beverage (therapeutic drug B) as described in Example 2, respectively. After intervention, the cell supernatant was collected from each group, and the LDH content in the supernatant was detected using a kit.

[0042] The specific method for determining the LDH content is as follows: The experimental equipment is shown in Table A. Table A

[0043] reagent kit composition Reagent 1: Matrix buffer, 5ml x 1 bottle, store at 2~8℃ for 6 months; Reagent 2: Coenzyme I, powder × 1 vial, store at 2~8℃ for 6 months; Preparation of Reagent 2 solution: Place Reagent 2 on an ice box, dissolve one vial of powder in 1.3 mL of double-distilled water, mix well, and store at 2-8℃ for 15 days. If multiple tests are required, aliquot and store at -20℃ for 2 weeks. Reagent 3: Phenyzine, 5mL x 1 bottle, store at 2~8℃ protected from light for 6 months; Reagent 4: NaOH, 5mL x 1 bottle, store at 2~8℃ for 6 months; Preparation of NaOH working solution: Mix according to the volume ratio of reagent 4:double distilled water 1:9. Use immediately after preparation. It can be stored at 2~8℃ for 7 days. Standard: 1 ml vial of 2 μmol / ml pyruvate standard, store at 2-8℃ for 6 months. Preparation of 0.2 μmol / ml pyruvate standard: Dilute the 2 μmol / ml pyruvate standard solution 10 times with double-distilled water, prepare fresh before use.

[0044] The operation steps and group processing for each group are shown in Table B; Table B

[0045] After adding the reagent to each group of cells, mix them gently, incubate at 37°C for 15 min, measure the absorbance (OD) value using an ELISA reader at a wavelength of 440 nm, and calculate cell viability. Calculation formula: LDH activity (U / L) = [(Model group OD - Blank group OD) / (Control group OD - Blank group OD)] * Standard group OD * N * 1000.

[0046] Note: N: Dilution factor of the sample before testing; C Standard: Standard solution concentration, 0.2 μmol / mL.

[0047] All data are expressed as mean ± standard deviation. The data were statistically analyzed using SPSS 19.0 software, employing a one-way ANOVA method. A p-value < 0.05 indicated a significant difference. GraphPad Prism 5.0 was used for plotting, and the results are shown in Table 5. Figure 4 As shown; Table 5. LDH activity analysis in cell supernatant of each group ( )

[0048] Note: △ Compared with the control group, P<0.05.

[0049] 2.3.3 Treatment Intervention Groups (1) Control group: Normal cultured cells; (2) Model group: DMEM without sugar and serum was used, and the cells were transferred to a three-gas incubator at 37°C (5% CO2, 94% N2, 1% O2). After 6 hours, the medium was replaced with normal medium, and the cells were transferred to a conventional incubator and cultured for another 12 hours. (3) Treatment group A: 24 hours before model construction, 40 mg / ml of treatment drug A was added, followed by glucose-free and serum-free DMEM, and the cells were transferred to a 37°C tri-gas incubator (5% CO2, 94% N2, 1% O2). After 6 hours, the medium was replaced with normal medium, and the cells were transferred to a conventional incubator for 12 hours of culture. (4) Treatment group B: 24 hours before model construction, 10 mg / ml of treatment drug B was added, followed by glucose-free and serum-free DMEM, and the cells were transferred to a 37°C tri-gas incubator (5% CO2, 94% N2, 1% O2). After 6 hours, the medium was replaced with normal medium, and the cells were transferred to a conventional incubator for 12 hours of culture. (5) Positive drug group: 24 hours before model construction, 10mM NAC (Acetylcysteine) was added, followed by sugar-free and serum-free DMEM, and the cells were transferred to a 37°C tri-gas incubator (5% CO2, 94% N2, 1% O2). After 6 hours, the medium was replaced with normal medium, and the cells were transferred to a conventional incubator for 12 hours of culture.

[0050] 2.3.4 Cell proliferation detection H9C2 cells with good growth and a confluence rate of 90% were collected and prepared into 5×10⁶ cells using complete culture medium. 4Cell suspension was seeded at 100 μL / well, 5 replicates per well. After 24 h of cell adhesion, cells were treated according to the experimental groupings in 2.3.3. After treatment, the culture medium was discarded, and 100 μL of prepared 10% CCK-8 solution was added to each well. Cells were incubated for another 1 h, and the OD value at 450 nm was measured using a microplate reader. All data are expressed as mean ± standard deviation. The data were statistically analyzed using SPSS 19.0 software, employing a one-way ANOVA method. A p-value < 0.05 indicated a significant difference. GraphPad Prism 5.0 was used for plotting, and the results are shown in Table 6. Figure 5 As shown; Table 6. Cell viability analysis of each group ( )

[0051] Note: △ Compared with the control group, P<0.05; ▲ Compared with the model group, P<0.05; ▽ Compared with treatment group A, P<0.05; ▼ Compared with treatment group B, P<0.05.

[0052] 2.3.5 LDH Content Detection H9C2 cells with good growth and a confluence rate of 90% were collected and prepared into 5×10⁶ cells using complete culture medium. 4 Single-cell suspensions of cells / mL were seeded into 24-well plates (500 μL / well, 3 replicates). After 24 h of cell adhesion, the cells were subjected to intervention according to the experimental groups in 2.3.3. After the intervention, the cell supernatant of each group was collected, and the LDH content in the supernatant was detected by ELISA kit. The ELISA detection method is a standard technique in this field, and the detection steps will not be described again. All data are expressed as mean ± standard deviation. The data were statistically analyzed using SPSS 19.0 software, employing a one-way ANOVA method. A p-value < 0.05 indicated a significant difference. GraphPad Prism 5.0 was used for plotting; the results are shown in Table 7. Figure 6 As shown; Table 7. LDH activity analysis in cell supernatant of each group ( )

[0053] Note: △ Compared with the control group, P<0.05; ▲ Compared with the model group, P<0.05; ▽ Compared with treatment group A, P<0.05; ▼ Compared with treatment group B, P<0.05.

[0054] The technical solutions provided by the embodiments of the present invention have been described in detail above. Specific examples have been used to illustrate the principles and implementation methods of the embodiments of the present invention. The descriptions of the embodiments above are only for helping to understand the principles of the embodiments of the present invention. At the same time, for those skilled in the art, there will be changes in the specific implementation methods and application scope based on the embodiments of the present invention. Therefore, the content of this specification should not be construed as a limitation of the present invention.

Claims

1. A tianxia apple drink, characterized in that: It includes the following components, by weight: 500-1000 parts of Hotan summer apple extract, 100-200 parts of Hotan red grape extract, 1-2 parts of Hotan rose, 0.1-0.5 parts of saffron, and 20-60 parts of flavoring agent.

2. A method of preparing the Yatsumi apple drink according to claim 1, characterized by: Includes the following steps: (1) Add water to the Hetian rose and heat and reflux to extract 2-3 times, 1-2 hours each time. Filter and combine the filtrates to form rose extract for later use. (2) Take saffron, add water, heat and reflux to extract 2-3 times, 1-2 hours each time, filter, and combine the filtrates to form saffron extract for later use; (3) Combine the extracts of Hetian summer apple, Hetian red grape, rose extract and saffron extract, add flavoring agent and mix thoroughly. Let stand at 0-4℃ for 12 hours, filter, bottle and sterilize.

3. The method for preparing the Hotan summer apple beverage according to claim 2, characterized in that: The Hotan summer apple extract is selected from fresh Hotan summer apple juice or dried Hotan summer apple water extract; the Hotan red grape extract is selected from fresh Hotan red grape juice or dried Hotan red grape water extract.

4. The method for preparing the Hotan summer apple beverage according to claim 3, characterized in that: The aqueous extract of Hetian summer apple is obtained by adding water to Hetian summer apple and heating and refluxing 2-3 times, each time for 1-2 hours, filtering, and combining the filtrates. The aqueous extract of Hetian red raisins is obtained by adding water to Hetian red raisins and heating and refluxing 2-3 times, each time for 1-2 hours, filtering, and combining the filtrates.

5. The method for preparing the Hotan summer apple beverage according to claim 2, characterized in that: The heating temperature in the heating reflux in step (1) is 90-100℃; the heating temperature in the heating reflux in step (2) is 95-100℃; the heating temperature in the heating reflux in step (3) is 90-100℃.

6. The use of the Hotan summer apple beverage according to claim 1 in the preparation of a drug for the prevention or treatment of myocardial injury.

7. The application according to claim 6, characterized in that: The drug is an oral medication.

8. The application according to claim 6, characterized in that: The drug also contains pharmaceutically acceptable excipients.

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