Composition for improving myocardial cell function and application thereof

Abstract

The invention discloses a composition for improving functions of myocardial cells and application of the composition. The composition comprises the following components: procyanidine, inulin, an astragalus extract and a seville orange flower extract. The composition belongs to the technical field of medicine and health care, all the components in the composition have a synergistic effect, the antioxidant effect can be comprehensively achieved, meanwhile, the intestinal function is improved, the constipation treatment effect is enhanced, the myocardial infarction area is remarkably reduced, and the curative effect is good. The myocardial cell function is comprehensively improved from the aspects of tonifying qi and blood, inhibiting oxygen free radicals, improving constipation and the like, and the compound can be applied to preparation of drugs for preventing or treating cardiovascular diseases such as myocardial infarction.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical and health care technology, specifically to a composition for improving myocardial cell function and its application. Background Technology

[0002] Myocardial protection is a crucial technique in cardiovascular treatment. Myocardial ischemia refers to a pathological state in which reduced blood perfusion to the heart leads to decreased oxygen supply, abnormal myocardial energy metabolism, and an inability to support normal cardiac function. The main clinical manifestations of ischemic heart disease include angina pectoris, myocardial infarction, and other ischemic symptoms. Oxygen free radicals generated during myocardial ischemia-reperfusion are a significant cause of myocardial cell damage and apoptosis. Oxygen free radicals possess strong oxidizing properties and can damage myocardial cells through various mechanisms, including lipid peroxidation, protein oxidation, DNA damage, and mitochondrial dysfunction. These mechanisms interact to contribute to the occurrence and development of myocardial ischemia and reperfusion injury. For patients with myocardial ischemia, traditional Chinese medicine treatment often focuses on promoting blood circulation, removing blood stasis, and nourishing qi and blood. Simultaneously, antioxidant effects can protect myocardial cell membranes, proteins, and DNA, maintaining normal cell function. Furthermore, clinical statistics show that constipation is prevalent in 40%-60% of patients with acute myocardial infarction. Straining during bowel movements not only increases the incidence of acute heart failure, potentially leading to sudden death in severe cases, but also significantly impacts patients' quality of life, causing immense physical and psychological suffering. Therefore, effectively preventing constipation in patients with myocardial infarction is also one of the key aspects of protecting the myocardium. Summary of the Invention

[0003] To address the shortcomings of existing technologies, this invention provides a composition for improving cardiomyocyte function and its application, which comprehensively improves cardiomyocyte function from multiple aspects such as "both qi and blood replenishment", inhibition of oxygen free radicals and improvement of constipation.

[0004] To achieve the objectives of this invention, the invention is implemented through the following technical solutions:

[0005] A composition for improving cardiomyocyte function, comprising, by weight: 6-20 parts of proanthocyanidins, 5-15 parts of inulin, 3-10 parts of astragalus extract, and 3-10 parts of bitter orange extract.

[0006] Furthermore, a composition for improving cardiomyocyte function, by weight, comprises: 12 parts proanthocyanidins, 10 parts inulin, 6 parts astragalus extract, and 8 parts bitter orange extract.

[0007] Furthermore, the preparation method of the Astragalus extract includes the following steps:

[0008] S1: Grind the astragalus root into powder;

[0009] S2: Add 10-15 times the amount of water to the pulverized Astragalus raw material from step S1 and decoct 1-2 times, then filter to obtain filtrate and residue.

[0010] S3: Take the filter residue from step S2, add 2-4 times the weight of water to the filter residue, sterilize, and then add 0.1-0.3% of activated Lactobacillus plantarum liquid to ferment; after fermentation, sterilize and filter to obtain fermentation liquid;

[0011] S4: Combine the filtrate from step S2 and the fermentation broth from step S3, centrifuge them, take the supernatant and concentrate it under reduced pressure to obtain a thick paste with a relative density of 1.1 to 1.3. Dry the thick paste under reduced pressure, pulverize it, and sieve it to obtain Astragalus extract.

[0012] The preparation method of the bitter orange flower extract includes the following steps:

[0013] S1: Crush the bitter orange flowers;

[0014] S2: Add 10-15 times the amount of water to the crushed bitter orange flower raw material in step S1 and decoct 1-2 times, then filter to obtain filtrate and residue.

[0015] S3: Take the filter residue from step S2, add 2-4 times the weight of water to the filter residue, sterilize, and then add 0.1-0.3% of activated Lactobacillus plantarum liquid to ferment; after fermentation, sterilize and filter to obtain fermentation liquid;

[0016] S4: Combine the filtrate from step S2 and the fermentation broth from step S3, centrifuge, and concentrate the supernatant under reduced pressure to obtain a thick paste with a relative density of 1.1 to 1.3. Dry the thick paste under reduced pressure, pulverize it, and sieve it to obtain the bitter orange extract.

[0017] Furthermore, the proanthocyanidins are preferably one or two of apple proanthocyanidins and grape seed proanthocyanidins.

[0018] Furthermore, the fermentation time of the Lactobacillus plantarum liquid is 48-72 hours, and the fermentation temperature is 30-37℃.

[0019] Furthermore, the present invention provides an oral formulation comprising the composition, wherein the oral formulation is prepared by adding conventional excipients to the composition and following conventional processes, and the oral formulation includes any one of powder, tablet, granule, capsule, solution, emulsion, and suspension.

[0020] Furthermore, the present invention provides the use of the composition or the formulation in the preparation of a medicament for the prevention or treatment of myocardial injury.

[0021] Furthermore, the myocardial injury is myocardial infarction.

[0022] The effects of each component in this invention are as follows:

[0023] Proanthocyanidins: Proanthocyanidins are powerful antioxidants. When there is local ischemia in the myocardium, proanthocyanidins can remove oxygen free radicals generated in the mitochondrial electron transport chain, thereby reducing tissue damage and helping the heart maintain normal function.

[0024] Inulin: Inulin is a soluble dietary fiber with functions such as regulating gut microbiota, anti-oxidation, anti-inflammation, and improving metabolism. Inulin can indirectly improve myocardial function by regulating the composition and function of gut microbiota. The antioxidant and anti-inflammatory effects of inulin have a protective effect on cardiomyocytes.

[0025] Astragalus extract: Astragalus has the effects of tonifying qi and strengthening the exterior, promoting diuresis and reducing swelling, and promoting tissue regeneration. Astragalus extract can reduce the content of harmful substances in the serum and myocardial tissue of rats with myocardial ischemia, and increase the content of energy metabolism substances in myocardial tissue, thereby alleviating myocardial ischemia damage. The effective components of astragalus, such as astragalus polysaccharides and astragalus saponins, can help dilate blood vessels and increase coronary blood flow, thereby improving the problem of insufficient myocardial oxygen supply. Astragalus also has good antioxidant effects, which can scavenge oxygen free radicals and reduce the damage of myocardial cells to oxidative stress caused by ischemia.

[0026] Bitter orange flower extract: Bitter orange flower has the effects of soothing the liver and relieving depression, regulating qi and harmonizing the stomach, promoting blood circulation and removing blood stasis, promoting diuresis and reducing swelling, moistening the intestines and relieving constipation, and anti-inflammatory and analgesic effects; it has the effect of regulating the cardiovascular system, can dilate blood vessels, and improve myocardial blood supply. Bitter orange flower extract can regulate the energy metabolism of myocardial cells and enhance the antioxidant capacity and energy supply of cells.

[0027] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0028] The composition of this invention, through the combined use of Astragalus membranaceus extract and Citrus aurantium extract, achieves dual replenishment of Qi and blood, conforming to the traditional Chinese medicine principle of "activating blood circulation and removing blood stasis, replenishing Qi and nourishing blood" in the treatment of myocardial ischemia. Extraction of Astragalus membranaceus and Citrus aurantium through combined decoction and fermentation methods ensures full extraction of effective components and enhances efficacy. Simultaneously, the addition of proanthocyanidins further enhances the antioxidant effect, effectively inhibiting oxygen free radicals, while the addition of inulin further improves the protective effect on cardiomyocytes and enhances the therapeutic effect on constipation. Furthermore, inulin, Astragalus membranaceus extract, and Citrus aurantium extract all possess certain antioxidant properties, indirectly slowing down the degradation of proanthocyanidins and enhancing the overall stability of the composition. This invention improves cardiomyocyte function from multiple aspects, including replenishing Qi and blood, inhibiting oxygen free radicals, and improving constipation. The components exhibit synergistic effects, comprehensively exerting antioxidant effects while improving intestinal function, enhancing the therapeutic effect on constipation, and significantly reducing the area of ​​myocardial infarction. It can be applied in the preparation of drugs for the prevention or treatment of cardiovascular diseases such as myocardial infarction. Attached Figure Description

[0029] Figure 1 The effect on the infarct area in mice with myocardial ischemia. Note: *p<0.05 compared with the model group; #p<0.05 compared with Example 1. Detailed Implementation

[0030] The present invention will now be clearly and completely described with reference to specific embodiments, but the present invention is not limited thereto. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention. Unless otherwise specified, the methods described are conventional methods. Unless otherwise specified, the materials described are all available from publicly available commercial sources.

[0031] Preparation Example 1:

[0032] The preparation method of Astragalus extract includes the following steps:

[0033] S1: Grind the astragalus root into powder;

[0034] S2: Add 12 times the amount of water to the crushed Astragalus raw material from step S1 and decoct twice, then filter to obtain filtrate and residue.

[0035] S3: Take the filter residue from step S2, add water at 3 times the weight of the filter residue, sterilize, and then add 0.2% of activated Lactobacillus plantarum P-8 bacterial solution (by weight of the filter residue) and ferment at 37°C for 48 hours; after fermentation, sterilize, filter, and obtain fermentation broth;

[0036] S4: Combine the filtrate from step S2 and the fermentation broth from step S3, centrifuge them, take the supernatant and concentrate it under reduced pressure to obtain a thick paste with a relative density of 1.1. Dry the thick paste under reduced pressure, pulverize it, and sieve it to obtain Astragalus extract.

[0037] The preparation method of bitter orange flower extract includes the following steps:

[0038] S1: Crush the bitter orange flowers;

[0039] S2: Add 10 times the amount of water to the crushed bitter orange flower raw material in step S1 and decoct twice, then filter to obtain filtrate and residue.

[0040] S3: Take the filter residue from step S2, add water at 3 times the weight of the filter residue, sterilize, and then add 0.3% of activated Lactobacillus plantarum P-8 bacterial solution (by weight of the filter residue) and ferment at 37°C for 56 hours; after fermentation, sterilize, filter, and obtain fermentation broth;

[0041] S4: Combine the filtrate from step S2 and the fermentation broth from step S3, centrifuge them, take the supernatant and concentrate it under reduced pressure to obtain a thick paste with a relative density of 1.2. Dry the thick paste under reduced pressure, pulverize it, and sieve it to obtain the bitter orange flower extract.

[0042] Example 1: A composition for improving cardiomyocyte function, the composition comprising: 12g grape seed proanthocyanidins, 10g inulin, 6g astragalus extract, and 8g bitter orange extract. The astragalus extract and bitter orange extract were prepared by the method of Example 1.

[0043] The composition is prepared by the following steps: each raw material is added to a high-speed mixer in sequence and mixed evenly.

[0044] Example 2: A composition for improving cardiomyocyte function, the composition comprising: 6g of apple proanthocyanidins, 5g of inulin, 3g of astragalus extract, and 3g of bitter orange extract. The astragalus extract and bitter orange extract were prepared by the method of Example 1.

[0045] The composition is prepared by the following steps: each raw material is added to a high-speed mixer in sequence and mixed evenly.

[0046] Example 3: A composition for improving cardiomyocyte function, the composition comprising: 10g each of apple proanthocyanidins and grape seed proanthocyanidins, 15g of inulin, 10g of astragalus extract, and 10g of bitter orange extract. The astragalus extract and bitter orange extract were prepared by the method of Example 1.

[0047] The composition is prepared by the following steps: each raw material is added to a high-speed mixer in sequence and mixed evenly.

[0048] Comparative Example 1: Compared with Example 1, glutathione was used instead of grape seed proanthocyanidins, and the amount of glutathione used was 12g.

[0049] Comparative Example 2: Compared with Example 1, fructooligosaccharides were used instead of inulin, and the amount of fructooligosaccharides used was 10g.

[0050] Comparative Example 3: Compared with Example 1, the composition lacked Astragalus extract and the amount of Citrus aurantium extract was adjusted to 14g.

[0051] Comparative Example 4: Compared with Example 1, the composition lacked bitter orange extract, and the amount of astragalus extract was adjusted to 14g.

[0052] Comparative Example 5: Compared with Example 1, the preparation steps of Astragalus membranaceus extract and Citrus aurantium extract lacked the S3 fermentation step.

[0053] Experimental Example

[0054] 1. Scavenging effect on DPPH free radicals: Samples from Examples 1-3 and Comparative Examples 1-5 were prepared at 250 μg / mL and centrifuged at 8000 r / min for 10 min. The supernatant was used as the test solution. A DPPH solution was prepared by mixing 0.1 mol / L DPPH stock solution with 95% ethanol. After adding the sample and mixing well, the solution was allowed to stand at room temperature in the dark for 30 min. After the reaction was completed, the absorbance was measured at 517 nm. The formula for calculating the DPPH free radical scavenging rate is: DPPH free radical scavenging rate (%) = [A0 - (A - A1)] / A0 × 100, where A0, A1, and A correspond to the absorbance values ​​of the blank well, control well, and test well, respectively.

[0055] As shown in Table 1, the present application achieves a DPPH free radical scavenging rate of over 67.18% through the combination of proanthocyanidins, inulin, astragalus extract, and bitter orange extract. Specifically, Example 1 achieved a DPPH free radical scavenging rate of 70.32%, which is superior to Comparative Example 1 (using glutathione instead of proanthocyanidins, 55.63%), Comparative Example 2 (using fructooligosaccharides instead of inulin, 60.42%), Comparative Example 3 (using bitter orange extract instead of astragalus extract, 48.84%), Comparative Example 4 (using astragalus extract instead of bitter orange extract, 56.79%), and Comparative Example 5 (lacking the *Lactobacillus plantarum* fermentation step in the preparation of astragalus extract and bitter orange extract, 45.26%). The antioxidant effects of using other antioxidants (glutathione) or dietary fibers (fructooligosaccharides) in the composition are relatively poor. The antioxidant effects of using only bitter orange extract or astragalus extract are also relatively poor. Furthermore, the antioxidant effects of the active ingredients obtained from astragalus extract and bitter orange extract by decoction alone are also relatively poor. These results indicate that the composition of this application, using proanthocyanidins as the antioxidant base, combined with inulin, astragalus extract, and bitter orange extract, which have certain antioxidant capabilities, and by optimizing the extraction methods of astragalus extract and bitter orange extract, can achieve better synergistic effects among the raw materials, resulting in stronger antioxidant activity, better scavenging of oxygen free radicals, and thus better protection of cardiomyocytes.

[0056] Table 1 Comparison of DPPH free radical scavenging rates among groups

[0057]

[0058] 2. Efficacy in treating constipation: Eight-week-old SPF-grade male Kunming mice were randomly divided into groups of 10 each. After 7 days of acclimatization, the mice were administered medication by gavage. The control group was given physiological saline by gavage daily, while the model group, positive control group, Example 1 group, and Comparative Examples 1-5 groups were all administered loperamide hydrochloride (10 mg / kg bw) by gavage to induce constipation. One hour later, the control group and model group were given an equal volume of physiological saline by gavage, the positive control group was given 0.5% phenolphthalein solution by gavage, and Example 1 and Comparative Examples 1-5 were given 0.5 g / kg body weight of the sample by gavage for 14 consecutive days. On days 0, 7, and 14, immediately after gavage, the mice were isolated in single cages for 3 hours, and feces were collected and the number of fecal particles was recorded.

[0059] As shown in Table 2, compared with the model group, the combination of proanthocyanidins, inulin, astragalus extract, and bitter orange extract in this application significantly increased the number of fecal particles in constipated mice (p < 0.05). The effect of improving constipation increased over time, reaching the improvement effect of the positive control group on day 14, and was significantly better than comparative examples 1-5. This indicates that the combination of the compositions in this application has a better effect. On the basis of improving constipation mainly with inulin, the combination with proanthocyanidins, astragalus extract, and bitter orange extract can further relieve constipation. Thus, while preventing and treating diseases such as myocardial ischemia and myocardial infarction, it can effectively improve constipation in patients and prevent the increased incidence of acute heart failure caused by straining during defecation.

[0060] Table 2. Number of fecal particles in each group of mice on days 0, 7, and 14 (3 hours).

[0061]

[0062]

[0063] Note: *p < 0.05 compared to the model group; #p < 0.05 compared to Example 1.

[0064] 3. Effect on the infarct area in mice with myocardial ischemia: Eight-week-old SPF-grade male Kunming mice were randomly divided into groups of 10 each. The mice were administered the drug by gavage. The blank control group and model group were given an equal volume of distilled water by gavage, while the positive control group was given 0.1 g / kg of Di'ao Xinxuekang by gavage. Examples 1 and Comparative Examples 1-5 were given 0.5 g / kg of body weight by gavage. The administration was once daily for 7 consecutive days, with free access to water and food during the treatment period. Except for the blank control group, the other groups received intraperitoneal injections of isoproterenol (60 mg / kg) for 3 consecutive days from day 8 to day 10 after gavage administration. After the mouse model was established, the mice were fasted, and their condition was recorded after 24 hours. The mice were then sacrificed, and their hearts were collected. The heart tissue was washed, cut vertically into 5 even slices, and stained in TTC staining solution at 37°C for 20 minutes. The slices were then fixed in 4% formaldehyde for 12 hours and photographed. Normal myocardium is stained red, while infarcted myocardium is white. Calculate the infarct area and the total myocardial area. Infarct area ratio = infarct area / total myocardial area × 100%.

[0065] like Figure 1 The results showed that, compared with the model group, the combination of proanthocyanidins, inulin, astragalus extract, and bitter orange extract in this application significantly reduced the proportion of myocardial ischemia-infarction area in mice, with a reduction effect close to that of the positive control group. Specifically, the proportion of myocardial infarction area in Example 1 was 27.39%, significantly lower than that in Comparative Example 1 (using glutathione instead of proanthocyanidins, 37.27%), Comparative Example 2 (using fructooligosaccharides instead of inulin, 35.53%), Comparative Example 3 (using bitter orange extract instead of astragalus extract, 40.93%), Comparative Example 4 (using astragalus extract instead of bitter orange extract, 38.71%), and Comparative Example 5 (lazybacterium plantarum fermentation step was missing in the preparation steps of astragalus extract and bitter orange extract, 42.59%). The above results demonstrate that, based on the principles of "promoting blood circulation and removing blood stasis, replenishing qi and nourishing blood" in traditional Chinese medicine treatment of myocardial ischemia, the combined use of Astragalus membranaceus extract and Citrus aurantium extract obtained through optimized extraction methods can achieve dual replenishment of qi and blood. At the same time, the combination of proanthocyanidins with antioxidant capacity and inulin, which can improve intestinal flora, allows the components to complement each other, further improving the function of myocardial cells and thus effectively preventing and treating diseases such as myocardial infarction.

[0066] The applicant declares that the above-described embodiments illustrate the products, uses, and methods of use of the present invention; however, the present invention is not limited to the detailed uses or methods of use described above, i.e., it does not mean that the present invention must rely on the detailed uses and methods of use described above to be realized. Those skilled in the art should understand that any improvements to the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A composition for improving cardiomyocyte function, characterized in that, The composition comprises, by weight, 6-20 parts of proanthocyanidins, 5-15 parts of inulin, 3-10 parts of astragalus extract, and 3-10 parts of bitter orange extract.

2. The composition for improving cardiomyocyte function according to claim 1, characterized in that, The composition comprises, by weight, 12 parts of proanthocyanidins, 10 parts of inulin, 6 parts of astragalus extract, and 8 parts of bitter orange extract.

3. A composition for improving cardiomyocyte function according to claim 1 or 2, characterized in that, The preparation method of the Astragalus extract includes the following steps: S1: Grind the astragalus root into powder; S2: Add 10-15 times the amount of water to the pulverized Astragalus raw material from step S1 and decoct 1-2 times, then filter to obtain filtrate and residue. S3: Take the filter residue from step S2, add 2-4 times the weight of water to the filter residue, sterilize, and then add 0.1-0.3% of activated Lactobacillus plantarum liquid to ferment; after fermentation, sterilize and filter to obtain fermentation liquid; S4: Combine the filtrate from step S2 and the fermentation broth from step S3, centrifuge them, take the supernatant and concentrate it under reduced pressure to obtain a thick paste with a relative density of 1.1 to 1.

3. Dry the thick paste under reduced pressure, pulverize it, and sieve it to obtain Astragalus extract.

4. A composition for improving cardiomyocyte function according to claim 1 or 2, characterized in that, The preparation method of the bitter orange flower extract includes the following steps: S1: Crush the bitter orange flowers; S2: Add 10-15 times the amount of water to the crushed bitter orange flower raw material in step S1 and decoct 1-2 times, then filter to obtain filtrate and residue. S3: Take the filter residue from step S2, add 2-4 times the weight of water to the filter residue, sterilize, and then add 0.1-0.3% of activated Lactobacillus plantarum liquid to ferment; after fermentation, sterilize and filter to obtain fermentation liquid; S4: Combine the filtrate from step S2 and the fermentation broth from step S3, centrifuge, and concentrate the supernatant under reduced pressure to obtain a thick paste with a relative density of 1.1 to 1.

3. Dry the thick paste under reduced pressure, pulverize it, and sieve it to obtain the bitter orange extract.

5. A composition for improving cardiomyocyte function according to claim 1 or 2, characterized in that, The proanthocyanidins are one or both of apple proanthocyanidins and grape seed proanthocyanidins.

6. A composition for improving cardiomyocyte function according to claim 3 or 4, characterized in that, The fermentation time of the Lactobacillus plantarum liquid is 48-72 hours, and the fermentation temperature is 30-37℃.

7. An oral preparation, characterized in that, The oral formulation includes the composition according to any one of claims 1-6, wherein the oral formulation comprises any one of powder, tablet, granule, capsule, solution, emulsion, and suspension.

8. The use of the composition according to any one of claims 1-6 or the formulation according to claim 7 in the preparation of a medicament for the prevention or treatment of myocardial injury.

9. The application according to claim 8, characterized in that, The myocardial injury mentioned is myocardial infarction.

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