Application of glycerophosphorylcholine to improvement of heart failure, heart remodeling and dysfunction

Abstract

The invention discloses application of glycerophosphorylcholine in improvement of heart failure, heart remodeling and dysfunction, and belongs to the technical field of biological medicine. The invention finds that the glycerophosphorylcholine can be used as a therapeutic drug for inhibiting development of heart failure and heart remodeling processes for the first time. In the heart remodeling process caused by the heart failure, the glycerophosphorylcholine can improve organic pathological changes of the heart, exogenous supplement of the glycerophosphorylcholine can effectively relieve myocardial cell hypertrophy, relieve the degree of cardiac fibrosis and restore heart remodeling related gene expression to the normal level, the effect on improving the heart remodeling is remarkable, and the composition has a good application prospect. The medicine can be used as a medicine for treating heart failure and heart remodeling, and the research result of the invention has great significance in effective intervention and treatment of heart failure and heart remodeling processes, and has huge application value in clinic.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical technology, and particularly relates to the application of glycerophosphocholine in improving heart failure, cardiac remodeling and dysfunction. Background Art

[0002] Heart failure is a clinical syndrome caused by cardiac dysfunction, and its incidence and mortality have been increasing year by year. Heart failure is a clinical syndrome caused by multiple factors. It is mainly due to the decline of cardiac systolic function, and the heart is insufficient to provide sufficient blood to meet the needs of the body, resulting in changes in myocardial structure and function. Heart failure is the end stage of various cardiovascular diseases. Hypertension and myocardial infarction are the main risk factors for heart failure. Its main manifestation is the decline of cardiac pumping ability, and it is unable to supply sufficient oxygen and nutrients required by various parts of the body. Cardiac remodeling is a key process in the deterioration of heart failure, which is specifically manifested in the abnormal hypertrophy of cardiomyocytes, involving an increase in the size of cardiomyocytes, that is, cardiac hypertrophy; on the other hand, it is manifested in the changes in cardiac structure and function.

[0003] Traditional methods for treating cardiac remodeling include drug therapy, surgical intervention and lifestyle adjustment, which can relieve symptoms and improve quality of life to a certain extent, but cannot fundamentally reverse the process of cardiac remodeling caused by heart failure or prevent the progression of the disease. Therefore, finding new therapeutic targets and developing new treatment strategies are of great significance for the treatment of cardiac remodeling and heart failure.

[0004] In recent years, researchers have paid increasing attention to the role of endogenous bioactive metabolite small molecules produced in vivo in heart diseases, especially those metabolic small molecules with the potential to regulate metabolism and cardiac function. Glycerophosphocholine (GPC), as an endogenous substance in the human body, is a biosynthetic precursor of the important neurotransmitter acetylcholine, and is considered to be one of the most suitable sources of choline to maintain the health of the human brain and psyche. At present, the research on GPC mostly focuses on mental diseases and is mainly used as a nutritional supplement for treating neurological diseases such as Alzheimer's disease and senile dementia. However, there is no relevant report on whether GPC has the effect of treating heart failure, cardiac remodeling and improving organic heart lesions. Summary of the Invention

[0005] In view of the above background introduction and the prior art, the object of the present invention is to provide the application of glycerophosphocholine (GPC) as a therapeutic drug in improving heart failure, cardiac remodeling and dysfunction. The present invention has found through research that the concentration of the bioactive metabolite small molecule GPC in the plasma of heart failure patients is reduced. During the progression of cardiac remodeling caused by heart failure, exogenous supplementation of GPC can significantly improve cardiomyocyte hypertrophy, reduce cardiac fibrosis, and restore the expression of genes related to cardiac remodeling caused by angiotensin II and ligation of the left anterior descending branch of the heart to normal levels. These results indicate that the bioactive metabolite small molecule GPC has good effects in treating cardiac remodeling and is expected to become a new target for the treatment of cardiac remodeling.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] In the first aspect of the present invention, there is provided the application of glycerophosphocholine in the preparation of a drug for improving heart failure.

[0008] The heart failure is caused by hypertension or myocardial infarction, and is specifically manifested as abnormal hypertrophy of cardiomyocytes and changes in cardiac structure.

[0009] The present invention has verified that there are significant differences in glycerophosphocholine in the plasma of heart failure patients and non-heart failure patients. The concentration level of glycerophosphocholine is positively correlated with the left ventricular ejection fraction and negatively correlated with myocardial injury and inflammation-related markers, providing the correlation between GPC and the risk of heart failure.

[0010] The detection of the GPC concentration is obtained by using plasma targeted metabolomics.

[0011] The glycerophosphocholine improves heart failure through at least one of the following (1) or (2) pathways:

[0012] (1) Alleviating the hypertrophy of cardiac tissue structure;

[0013] (2) Reducing the cross-sectional area of cardiomyocytes and reducing the degree of cardiac fibrosis.

[0014] In the second aspect of the present invention, there is provided the application of glycerophosphocholine in the preparation of a drug for improving cardiac remodeling.

[0015] The cardiac remodeling is induced by angiotensin II and ligation of the left anterior descending branch, and cardiac remodeling is a key process in the deterioration of heart failure.

[0016] The in vivo models of heart failure and cardiac remodeling constructed by the present invention are induced by angiotensin II and ligation of the left anterior descending branch of the heart; the in vitro model of cardiac remodeling is induced by culturing cardiomyocytes under the conditions of angiotensin II and hypoxia.

[0017] The choline glycerophosphate improves cardiac remodeling through at least one of the following pathways (1)-(3):

[0018] (1) Alleviating the hypertrophy of cardiac tissue structure;

[0019] (2) Reducing the cross-sectional area of cardiomyocytes and alleviating the degree of cardiac fibrosis;

[0020] (3) Downregulating the upregulation of gene expressions related to cardiac hypertrophy and fibrosis.

[0021] The genes related to cardiac hypertrophy and fibrosis are: natriuretic peptide a gene, natriuretic peptide b gene, and type III collagenase gene.

[0022] Advantages of the present invention:

[0023] The present invention for the first time discovers that choline glycerophosphate can be used as a therapeutic drug to inhibit the development of heart failure and cardiac remodeling processes. During the cardiac remodeling process caused by heart failure, choline glycerophosphate can improve the organic lesions of the heart. Exogenous supplementation of choline glycerophosphate can effectively alleviate cardiomyocyte hypertrophy, reduce the degree of cardiac fibrosis, and restore the gene expressions related to cardiac remodeling to normal levels. It has a significant effect in improving cardiac remodeling and can be used as a drug for treating heart failure and cardiac remodeling. The research results of the present invention are of great significance for the effective intervention and treatment of heart failure and cardiac remodeling processes and have great application value in clinical practice. Description of the Drawings

[0024] Figure 1 It shows the distribution of two groups of patients.

[0025] Figure 2 It shows the concentrations of choline glycerophosphate (GPC) in the plasma of two groups of patients.

[0026] Figure 3 It shows the correlation analysis of GPC concentration with markers related to myocardial injury and inflammation.

[0027] Figure 4 It shows the correlation of GPC concentration with left ventricular ejection fraction.

[0028] Figure 5 It shows the results and analysis of cardiac echocardiography Doppler of each group of mice. Among them, Figure 5 a in it shows the results of cardiac echocardiography Doppler, Figure 5 b in it shows the detection of left ventricular ejection fraction of each group, Figure 5 c in it shows the detection of left ventricular systolic function (E / E’) of each group, Figure 5 d in it shows the detection of heart mass / tibiofemoral length of each group, Figure 5 e in it shows the detection of heart mass / body weight of each group.

[0029] Figure 6Hematoxylin-eosin staining images of cardiac tissues for each group.

[0030] Figure 7 Images of wheat germ agglutinin (WGA) staining of cardiac tissues for each group and statistics of cross-sectional area of cardiomyocytes. Among them, Figure 7 a in it is WGA staining. Figure 7 b in it is statistics of cross-sectional area of cardiomyocytes.

[0031] Figure 8 Images of Masson staining of cardiac tissues for each group and statistics of fibrosis degree. Among them, Figure 8 a in it is Masson staining. Figure 8 b in it is statistics of fibrosis degree.

[0032] Figure 9 Expression levels of mRNAs related to cardiac hypertrophy and fibrosis in cardiac tissues of mice in each group.

[0033] Figure 10 Analysis images of cardiomyocyte hypertrophy (β-actin staining) and mitochondrial morphology (Mitotracker Deep Red) in a pathological myocardial remodeling model constructed by angiotensin II-induced AC16 cells. Among them, Figure 10 a in it is the image of mitochondrial morphology. Figure 10 b in it is statistics of cardiomyocyte area. Figure 10 c in it is the staining image of cardiomyocytes.

[0034] Figure 11 Results and analysis of echocardiography Doppler in the hearts of mice in each group. Among them, Figure 11 a in it is the result of echocardiography Doppler. Figure 11 b in it is statistics of left ventricular ejection fraction. Figure 11 c in it is statistics of left ventricular shortening fraction. Figure 11 d in it is statistics of left ventricular volume at end-systole. Figure 11 e in it is statistics of left ventricular volume at end-diastole.

[0035] Figure 12 Hematoxylin-eosin staining images of cardiac tissues for each group.

[0036] Figure 13 Images of wheat germ agglutinin (WGA) staining of cardiac tissues for each group and cross-sectional area of cardiomyocytes. Among them, Figure 13 a in it is the image of WGA staining. Figure 13 b in it is statistics of cross-sectional area of cardiomyocytes.

[0037] Figure 14 Sirius red staining and fibrosis degree of cardiac tissues for each group. Among them, Figure 14 a in it is Sirius red staining. Figure 14 b in it is statistics of fibrosis degree.

[0038] Figure 15 For the expression levels of mRNAs related to myocardial hypertrophy and fibrosis in the heart tissues of each group of mice, where Figure 15 a in it is the expression level of the ANP gene, Figure 15 b in it is the expression level of the ColⅢ gene.

[0039] Figure 16 To establish a myocardial infarction model by hypoxic induction of AC16 cells and detect the changes in myocardial cell hypertrophy (β-actin staining).

[0040] Figure 17 To establish a myocardial infarction model by hypoxic induction of AC16 cells and detect the changes in mitochondrial morphology (Mitotracker Deep Red). Detailed implementation manners

[0041] It should be noted that the following detailed description is illustrative and aims to provide further explanation for the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which the present application belongs.

[0042] As mentioned above, heart failure is a clinical syndrome caused by cardiac dysfunction, and its incidence shows an increasing trend year by year. Cardiac remodeling is a key process leading to the deterioration of heart failure. The specific manifestations of cardiac remodeling are the enlargement of myocardial cells and the changes in the structure and function of the heart. Therefore, actively developing new treatment strategies is of great significance for the treatment of heart failure and cardiac remodeling.

[0043] Based on this, the present invention provides the application of glycerophosphocholine in improving cardiac structure and function disorders caused by heart failure and cardiac remodeling. According to the definition of heart failure in "Diagnosis and Treatment of Acute and Chronic Heart Failure by the European Society of Cardiology 2022", heart failure patients and non-heart failure patients were included. Through targeted metabolomics research on the plasma of heart failure patients and non-heart failure patients, glycerophosphocholine showed a significant change trend, and its level in the heart failure population was significantly decreased. During the occurrence and development of heart failure, the aggravation of cardiac function damage is often accompanied by the increase of myocardial injury and inflammation-related markers and the decrease of left ventricular ejection fraction (LVEF%). According to the analysis of clinical test results, the level of glycerophosphocholine was positively correlated with the left ventricular ejection fraction and negatively correlated with myocardial injury and inflammation-related markers. Therefore, glycerophosphocholine can be used as a beneficial substance and as a therapeutic drug for the treatment of heart failure and the intervention of the cardiac remodeling process.

[0044] To confirm that the level of glycerophosphocholine is significantly reduced in patients with heart failure and whether exogenous supplementation can alleviate cardiac hypertrophy and the degree of damage to cardiac structure and function, we further constructed a mouse disease model and judged the effect of exogenous supplementation of glycerophosphocholine on cardiac remodeling. The gene expression levels of natriuretic peptide a (ANP) and natriuretic peptide b (BNP) in mouse heart tissue mark the degree of myocardial hypertrophy in the heart tissue of mice in different experimental groups, and the gene expression level of type III collagenase (ColIII) in mouse heart tissue marks the degree of fibrosis in the heart tissue of mice in different experimental groups. This invention confirmed through animal experiments that glycerophosphocholine can significantly alleviate cardiac hypertrophy caused by angiotensin II and ligation of the left anterior descending branch of the heart, reduce myocardial fibrosis caused by heart failure, and can significantly reverse the up-regulated genes related to cardiac hypertrophy and fibrosis, and better alleviate the process of cardiac remodeling.

[0045] The following further describes in detail the specific implementation manners of the present invention in conjunction with embodiments. The following detailed descriptions are all illustrative and are intended to provide further descriptions of this application rather than limiting the scope of the present invention.

[0046] Unless otherwise specified, the reagents and equipment used in the embodiments of the present invention are all conventional reagents and conventional equipment in the technical field, and the CAS of the glycerophosphocholine is: 28319-77-9.

[0047] Example 1: Correlation between glycerophosphocholine and the risk of heart failure

[0048] A total of 1434 inpatients clinically diagnosed with ischemic heart disease were included in this example. The cardiac function was evaluated according to the left ventricular ejection fraction (LVEF%), and the cardiac structure parameters were obtained by echocardiography. According to the definition of heart failure in the "2022 European Society of Cardiology Guidelines for the Diagnosis and Treatment of Acute and Chronic Heart Failure", the above patients were divided into a heart failure group (HF) and a non-heart failure group (non-HF). The proportion of each population is shown in Figure 1 as shown.

[0049] By detecting the plasma of the two groups of patients by targeted metabolomics and quantitatively analyzing and comparing the concentrations of glycerophosphocholine GPC in the plasma of the two groups of patients ( Figure 2 ), the results showed that: compared with non-heart failure patients, the concentration of GPC in the plasma of heart failure patients decreased significantly.

[0050] Subsequently, a correlation analysis was performed on the GPC level and markers related to myocardial injury and inflammation ( Figure 3 ), and the results showed that the level of GPC was negatively correlated with markers related to myocardial injury and inflammation. In addition, the level of GPC was positively correlated with the left ventricular ejection fraction LVEF% ( Figure 4) That is, GPC plays an important protective role in heart failure and cardiac remodeling and can be used as a therapeutic drug.

[0051] Example 2: Therapeutic effect of glycerophosphocholine on a mouse model of angiotensin II-induced pathological myocardial remodeling To evaluate the effect of glycerophosphocholine (GPC) on myocardial hypertrophy in vivo, the following experiment was carried out:

[0052] (1) Experimental method:

[0053] Twenty healthy 8-week-old male C57BL / 6 mice were selected and divided into a control group (Saline + Saline), a model group (AngⅡ + Saline), a low-dose group (AngⅡ + m-GPC), and a high-dose group (AngⅡ + h-GPC), with 5 mice in each group.

[0054] Mice in the control group were implanted with a micro-osmotic pump (Alzet model 1002) under 2% isoflurane anesthesia and continuously administered saline (Saline) at a rate of 1.0 mg / kg / d through the Alzet micro-osmotic pump, and saline was administered once a day by intraperitoneal injection (I.P.). The dosing volume V (μL) of saline = m (g) * 5.

[0055] Mice in the model group were implanted with a micro-osmotic pump (Alzet model 1002) under 2% isoflurane anesthesia and continuously administered angiotensin II (AngII) at a rate of 1.0 mg / kg / d through the Alzet micro-osmotic pump, and saline was administered once a day by intraperitoneal injection (I.P.). The dosing volume of saline was the same as that of the control group.

[0056] Mice in the low-dose group (m-GPC, 10 mg / kg / d) were implanted with a micro-osmotic pump (Alzet model 1002) under 2% isoflurane anesthesia and continuously administered angiotensin II (AngII) at a rate of 1.0 mg / kg / d through the Alzet micro-osmotic pump, and glycerophosphocholine (GPC) was administered once a day by intraperitoneal injection (i.p.). A GPC stock solution of 2 mg / mL was prepared, and the dosing volume was calculated based on body weight: GPC dosing volume V (μL) = m (g) * 5.

[0057] Mice in the high-dose group (h-GPC, 20 mg / kg / d) were implanted with a micro-osmotic pump (Alzet model 1002) under 2% isoflurane anesthesia, and angiotensin II (AngII) was continuously administered at a rate of 1.0 mg / kg / d through the Alzet micro-osmotic pump. Glycerophosphocholine GPC was administered once a day via intraperitoneal injection (I.P.). A GPC stock solution of 4 mg / ml was prepared, and the dosing volume was calculated based on body weight: GPC dosing volume V (μL) = m (g) * 5.

[0058] After two weeks, the mice were anesthetized with ketamine (at a ratio of 10 mg / kg), and then cardiac Doppler ultrasound was performed to compare changes in cardiac structure and function. Finally, the mouse hearts were removed for quantitative analysis experiments such as histological staining analysis and real-time quantitative polymerase chain reaction for related gene expression analysis.

[0059] Among them, the wheat germ agglutinin staining (WGA) experiment of cardiac tissue includes the following steps:

[0060] 1) Baking and dewaxing: Place the paraffin sections of cardiac tissue in an oven at 60 °C for 1 h to melt the paraffin and prevent de-sloughing. Then, successively incubate in xylene I, xylene II, and xylene III for 5 minutes each; and in absolute ethanol, 95% ethanol, 85% ethanol, and 75% ethanol for 5 minutes each to dewax to the aqueous phase. After dewaxing, rinse with running water for 10 minutes;

[0061] 2) Dropwise add the WGA working solution (1:200) and incubate at 37 °C in the dark for 1 hour;

[0062] 3) Wash with PBS 3 times, 3 minutes each time. Mount the sections with an anti-fluorescence quenching mounting medium, being careful not to generate bubbles. After air-drying, take pictures.

[0063] The hematoxylin-eosin staining experiment includes the following steps:

[0064] 1) After baking and dewaxing (the same steps as WGA staining), stain the nuclei with hematoxylin for 5 minutes, then differentiate with hydrochloric acid alcohol differentiation solution for 5 seconds, and then quickly rinse in slow running water for 10 minutes to blue with the substances in the tap water;

[0065] 2) Stain the cytoplasm with eosin for 5 minutes. After washing off the floating color with tap water, quickly start dehydration and clearing. The dehydration and clearing process is successively carried out in 75% ethanol, 85% ethanol, 95% ethanol, and absolute ethanol for 5 minutes each; and in xylene I, xylene II, and xylene III for 5 minutes each;

[0066] 3) Mounting: Quickly mount with neutral balsam. After air-drying, take pictures.

[0067] The Masson trichrome staining experiment includes the following steps:

[0068] 1) After baking the slices and dewaxing (the same steps as in WGA staining), place them in Bouin's solution at 4 °C overnight. After overnight incubation, rinse the tissue sections with tap water until the yellow color fades.

[0069] 2) Stain with Ponceau S for 30 minutes. Drop double-distilled water above the glass slide to let the water flow downward to wash off the Ponceau S. After differentiating with phosphomolybdic acid for 1 minute, stain with aniline blue for 3 minutes. Similarly, drop double-distilled water above the glass slide to let the water flow downward to wash off the aniline blue.

[0070] 3) After dehydration and clearing (the same as in hematoxylin-eosin staining), quickly mount the sections with neutral balsam. After air-drying, take pictures.

[0071] (2) Experimental results:

[0072] The results of cardiac ultrasound Doppler ( Figure 5 ) showed that compared with the control group, the left ventricular ejection fraction EF% of the mice in the model group was significantly decreased, the cardiac systolic function declined. After exogenous supplementation of GPC, EF% was significantly increased ( Figure 5 b); the value of E / E' of the mice in the model group was significantly increased and significantly decreased after exogenous supplementation of GPC ( Figure 5 c); the ratio of heart weight (HW, g) to body weight (BW, g) and the ratio of heart weight (HW, g) to tibia length (TL, cm) of the mice in the model group were significantly increased and significantly decreased after exogenous supplementation of GPC ( Figure 5 d and Figure 5 e).

[0073] The hematoxylin-eosin staining images of cardiac tissues of different groups ( Figure 6 ), the schematic diagrams of the cross-sectional area of cardiomyocytes in cardiac tissues ( Figure 7 ), and the Masson staining images of cardiac tissues of different groups ( Figure 8 ) showed that glycerophosphocholine GPC could relieve the cardiac tissue structure hypertrophy induced by angiotensin II, reduce the cross-sectional area of cardiomyocytes, and alleviate the level of cardiac fibrosis. The analysis of the expression levels of ANP and BNP genes reflecting the degree of cardiac hypertrophy in cardiac tissues of mice in different groups and the expression level of collagenase III (ColⅢ) gene in cardiac tissues ( Figure 9 ) showed that the supplementation of high-concentration glycerophosphocholine GPC reversed the changes in the expression of genes related to cardiac remodeling induced by angiotensin II, prevented the hypertrophy of cardiomyocytes induced by angiotensin II, and alleviated the up-regulation of genes related to cardiac remodeling.

[0074] Example 3: Protective effect of glycerophosphocholine on cardiomyocyte cell lines in vitro in simulating cardiac hypertrophy

[0075] (1) Experimental method:

[0076] The culture of AC16 human cardiomyocytes was carried out in F12 / DMEM medium containing 10% fetal bovine serum under the conditions of 37 °C and 5% CO2. After the AC16 human cardiomyocytes were inoculated into a 12-well cell culture plate, they were first cultured normally until they reached an appropriate density. To evaluate the effect of glycerophosphocholine (GPC) on myocardial hypertrophy in vitro, the AC16 human cardiomyocytes were divided into a control group (Control), a GPC treatment group (12 μg / mL), a model group (Ang II, 1 μM), and an experimental group (Ang II + 12 μg / mL GPC). In the model group, the cells were treated with 1 μM Ang II for 48 hours; in the experimental group, the cells were first treated with 1 μM Ang II for 24 hours, and then co-treated with 1 μM Ang II and 12 μg / mL GPC for 24 hours; the control group was cultured normally; the GPC treatment group was treated with 12 μg / mL GPC for 24 hours. Subsequently, the cardiomyocytes were stained with mitochondria (MitoTracker Deep Red, 100 nM) and observed, and β-actin staining was used for quantitative analysis of cell area.

[0077] (2) Experimental results:

[0078] In addition, in vitro experiments, the β-actin staining results and mitochondrial staining results of human cardiomyocytes AC16 showed that glycerophosphocholine (GPC) could alleviate cardiomyocyte hypertrophy and mitochondrial damage caused by angiotensin II ( Figure 10 ).

[0079] Example 4: Therapeutic effect of glycerophosphocholine on myocardial infarction mouse model

[0080] (1) Experimental method:

[0081] Healthy 8-week-old male C57BL / 6 mice were selected and divided into a sham operation group (Sham, saline, 6 mice), a model group (MI, saline, 6 mice), and a treatment group (MI + GPC, 10 mg / kg / d, 7 mice).

[0082] The control group of mice was anesthetized with 2% isoflurane and a control Sham group was constructed by sham operation, and saline was administered daily by intraperitoneal injection (i.p.), and the dosage V (μL) = m (g) * 5.

[0083] The model group of mice was anesthetized with 2% isoflurane and a myocardial infarction model was constructed by ligation of the left anterior descending branch, and saline was administered daily by intraperitoneal injection (i.p.), and the dosage was the same as that of the control group.

[0084] The mice in the treatment group (MI + GPC, 10 mg / kg / d) were anesthetized with 2% isoflurane and a myocardial infarction model was constructed by ligating the left anterior descending branch. Glycerophosphocholine (GPC, 2 mg / mL) was administered daily via intraperitoneal injection (i.p.), and the dosage volume V (μL) = m (g) * 5.

[0085] After 21 days, the mice were anesthetized with ketamine (at a ratio of 10 mg / kg). After anesthesia, cardiac Doppler ultrasound was performed to compare the changes in the cardiac structure and function of the mice. Finally, the mouse hearts were removed for quantitative analysis experiments such as histological staining analysis and real-time quantitative polymerase chain reaction for related gene expression analysis.

[0086] Among them, the experimental methods of wheat germ agglutinin staining (WGA) and hematoxylin-eosin staining of cardiac tissue were the same as those described in Example 2.

[0087] The Picrosirius red (picric acid Sirius red staining) experiment included the following steps:

[0088] 1) After baking and dewaxing the sections (the steps are the same as above), the sections were placed in a 0.1% picric acid Sirius red staining solution for 1 h and rinsed with running tap water for 5 min;

[0089] 2) After dehydration and clearing (the same as hematoxylin-eosin staining), the sections were quickly sealed with neutral balsam. After drying, photographs were taken.

[0090] (2) Experimental results:

[0091] The results of cardiac ultrasound Doppler ( Figure 11 ) showed that compared with the sham operation group, the left ventricular ejection fraction and left ventricular fractional shortening of the mice in the model group were significantly reduced, indicating a decline in cardiac systolic function. After exogenous supplementation of GPC, the left ventricular ejection fraction and left ventricular fractional shortening were significantly increased ( Figure 11 b - c); the left ventricular volume in the systolic and diastolic phases of the mice in the model group was significantly increased, indicating a decline in cardiac diastolic function, and it was significantly decreased after exogenous supplementation of GPC ( Figure 11 d - e).

[0092] The hematoxylin-eosin staining images of cardiac tissue in different groups ( Figure 12 ), the schematic diagrams of the cross-sectional area of cardiomyocytes in cardiac tissue in different groups ( Figure 13 ), and the Picrosirius staining images at the interstitium of cardiac tissue in different groups ( Figure 14 ) indicated that glycerophosphocholine (GPC) could relieve the cardiac tissue structure hypertrophy induced by myocardial infarction, reduce the cross-sectional area of cardiomyocytes, and alleviate the degree of cardiac fibrosis.

[0093] Analysis of the expression levels of ANP gene reflecting the degree of myocardial hypertrophy in the heart tissues of mice in different groups and collagenase III (ColIII) gene in the heart tissues Figure 15 The detection results show that the supplementation of high-concentration glycerophosphocholine (GPC) reverses the changes in the expression of genes induced by myocardial infarction, prevents myocardial cell hypertrophy induced by myocardial infarction, and reduces the upregulation of genes related to cardiac remodeling.

[0094] Example 5: Protective effect of glycerophosphocholine on cardiomyocyte cell line in a simulated myocardial infarction model

[0095] (1) Experimental method:

[0096] After AC16 human cardiomyocytes were inoculated into a 12-well cell culture plate, they were first cultured normally until they reached an appropriate density. To evaluate the effect of glycerophosphocholine (GPC) on myocardial hypertrophy in vitro, AC16 human cardiomyocytes were divided into a control group (normoxia + Control), a GPC treatment group (normoxia + 12 μg / mL GPC), a model control group (hypoxia), and a model treatment group (hypoxia + 12 μg / mL GPC). Among them, the cells in the model control group were treated with hypoxia in a 1% oxygen incubator for 24 hours; the cells in the model treatment group were treated with hypoxia in a 1% oxygen incubator for 24 hours and co-treated with 12 μg / mL GPC for 24 hours at the same time; the control group and the GPC treatment group were both cultured normally, and the GPC treatment group was treated with 12 μg / mL GPC for 24 hours. Subsequently, the cardiomyocytes were stained with mitochondria (MitoTracker Deep Red, 100 nM) and observed, and β-actin staining was used for quantitative analysis of cell area.

[0097] (2) Experimental results:

[0098] In addition, in vitro experiments, the β-actin staining results and mitochondrial staining results of human cardiomyocytes AC16 showed that glycerophosphocholine (GPC) could relieve myocardial cell hypertrophy caused by hypoxia Figure 16 ) and mitochondrial damage Figure 17 ).

[0099] The above are only the preferred embodiments of the present application and are not used to limit the present application. For those skilled in the art, the present application can have various changes and modifications. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. Use of glycerophosphocholine in the preparation of a drug for improving heart failure.

2. The application according to claim 1, characterized in that, The heart failure is caused by hypertension or myocardial infarction.

3. The application according to claim 1, characterized in that, The glycerophosphocholine improves heart failure through at least one of the following ways (1) or (2): (1) Alleviating cardiac tissue structure hypertrophy; (2) Reducing the cross-sectional area of cardiomyocytes and alleviating the degree of cardiac fibrosis.

4. Use of glycerophosphocholine in the preparation of a drug for improving cardiac remodeling.

5. The application according to claim 4, wherein The cardiac remodeling is induced by angiotensin II and left anterior descending coronary artery ligation.

6. The application according to claim 4, wherein The glycerophosphocholine improves cardiac remodeling through at least one of the following ways (1)-(3): (1) Alleviating cardiac tissue structure hypertrophy; (2) Reducing the cross-sectional area of cardiomyocytes and alleviating the degree of cardiac fibrosis; (3) Down-regulating the up-regulation of genes related to cardiac hypertrophy and fibrosis.

7. The application according to claim 6, wherein The genes related to cardiac hypertrophy and fibrosis are: natriuretic peptide a gene, natriuretic peptide b gene and type III collagenase gene.

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