Application of glossy privet fruit glycoside in preparation of medicine for treating senile cardiac hypofunction

Abstract

The invention discloses application of glossy privet fruit glycoside in preparation of a medicine for treating senile cardiac hypofunction, belongs to the technical field of medicines, and particularly relates to the application of the glossy privet fruit glycoside in preparation of the medicine for treating senile cardiac hypofunction. The polypeptide can effectively improve myocardial hypertrophy and cardiac function impairment of old mice, reduce myocardial fibrosis, reduce inflammation and senescence marker levels, and have potential application value in anti-aging and heart protection aspects.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and more particularly to the use of ligustroside in the preparation of a medicament for treating senile cardiac hypofunction. Background Art

[0002] Heart failure (HF) is a prevalent disease with a poor prognosis, and age is one of its major risk factors. With the significant increase in the trend of population aging, it has become crucial to understand how the biological aging process affects HF at the mechanical level. Senile heart failure is a common cardiovascular disease in the elderly. However, the physiological characteristics of the elderly population itself and the changes in the cardiovascular system pose challenges to the diagnosis and treatment of heart failure. Different from the young population, elderly heart failure patients have characteristics such as natural decline of body functions, coexistence of multiple cardiovascular disease risk factors, co-morbidity of multiple diseases, and polypharmacy. Due to the degenerative changes of the heart, the cardiac reserve function of elderly heart failure patients is weakened, the systolic function is reduced, and the overall myocardial compliance decreases, resulting in ventricular diastolic and filling disorders, increased left atrial pressure and pulmonary vascular pressure. In addition, in the state of heart failure, due to reduced respiratory function, pulmonary congestion, low cardiac output, abnormal distribution of pulmonary blood flow and gas exchange, elderly patients are more likely to develop hypoxemia. Moreover, the elderly population often coexists with multiple comorbidities, such as hypertension, diabetes, chronic renal insufficiency, chronic obstructive pulmonary disease, anemia, etc. More than 75% of elderly heart failure patients have 3 or more diseases, and 50% of patients have 5 or more diseases. Considering the characteristics of the elderly population such as coexistence of multiple diseases, polypharmacy, and natural decline of body functions, the overall management of heart failure in the elderly faces more severe difficulties than the general population.

[0003] Ligustroside is an active ingredient in olives and plays an important role in the Mediterranean diet. In early studies, it was thought that oleuropein in olive oil played an important role. Through continuous in-depth research, it was found that the hydrolysis product of oleuropein, ligustroside, has stronger physiological activity. It has a simple structure, a small molecular weight, good water solubility and lipid solubility, can prevent damage to the retina caused by radiation, oxygen free radicals and other factors, and has good therapeutic and health care functions, which has attracted extensive attention from researchers at home and abroad. Although it has been reported in the prior art that neo-ligustroside has various biological activities, the application of ligustroside in drugs for improving cardiac hypofunction has not been reported so far.

[0004] In view of this, the inventor of the present invention conducted in-depth research on this need, and thus this case was born. Summary of the Invention

[0005] The present invention aims to provide the application of ligustroside in the preparation of drugs for treating senile cardiac hypofunction, which can effectively improve myocardial hypertrophy and impaired cardiac function in senile mice, reduce myocardial fibrosis, and lower the levels of inflammation and senescence markers, demonstrating its potential application value in anti-aging and cardiac protection.

[0006] The present invention provides the application of ligustroside in the preparation of drugs for treating senile cardiac hypofunction. Specifically, the drugs for treating senile cardiac hypofunction include ligustroside as an active ingredient.

[0007] Preferably, in the drugs for treating senile cardiac hypofunction, ligustroside is used as the sole active ingredient or the main active ingredient.

[0008] Preferably, the application of ligustroside in the preparation of the drugs for treating senile cardiac hypofunction in the senescent state.

[0009] Preferably, the senescent state includes any one of the senescent states caused by the natural decline of body functions, coexistence of cardiovascular diseases, or multiple medications.

[0010] Preferably, the cardiac hypofunction includes myocardial fibrosis, cardiac function injury, and myocardial hypertrophy.

[0011] The present invention also provides a drug for treating senile cardiac hypofunction, which includes ligustroside and pharmaceutically acceptable auxiliary components.

[0012] Preferably, the dosage form of the drug for treating senile cardiac hypofunction includes powder, granule, tablet, capsule, pill, solution, suspension, or injection.

[0013] The beneficial effects produced by the technical solution of the present invention are as follows:

[0014] Through the present invention, ligustroside is proven to be able to effectively improve myocardial hypertrophy and impaired cardiac function in senile mice, reduce myocardial fibrosis, and lower the levels of inflammation and senescence markers, indicating its potential application value in anti-aging and cardiac protection. Since the application of ligustroside in the preparation of drugs for treating senile cardiac hypofunction is the first discovery of the inventor, therefore, whether ligustroside is used alone as an active ingredient to make a medicament, or used in combination with other active ingredients of ligustroside to make a medicament, it is within the protection scope of this patent. Description of the Drawings

[0015] To more clearly illustrate the technical solutions of the embodiments of the present invention, the following will briefly introduce the drawings required for the embodiments. It should be understood that the following drawings only show some embodiments of the present invention, and therefore should not be regarded as a limitation of the scope. For those of ordinary skill in the art, without creative efforts, other related drawings can also be obtained based on these drawings.

[0016] Figure 1 It is a comparison diagram of the cross-sectional areas of cardiomyocytes in each group of mice in Experiment 1;

[0017] Figure 2 It is a comparison diagram of the expression levels of inflammatory factors IL-6 and TNF in each group of mice in Experiment 1;

[0018] Figure 3 It is a comparison diagram of the expression levels of cardiac function impairment markers in the hearts of each group of mice in Experiment 1;

[0019] Figure 4 It is a data analysis diagram of RNA sequences under different cardiac pathological states in Experiment 2;

[0020] Figure 5 It is a scatter plot of CHIP signals in macrophages and monocytes in Experiment 2;

[0021] Figure 6 It is a scatter plot of the mRNA expression levels of CHIP signals and age growth, myocardial stress marker (NPPB), myocardial fibrosis marker (ACTA2), and inflammatory indicators (IL-6, IL-1β) in Experiment 2;

[0022] Figure 7 It is a diagram of the expression level of the senescence marker P21 in the heart tissue of mice in the CHIP model group in Experiment 2;

[0023] Figure 8 It is a staining diagram of the myocardial fibrosis area in each group of mice in Experiment 2;

[0024] Figure 9 It is a diagram of the transcriptional factor level expression of senescence markers S100A8, S100A9, and SPP1 in the heart tissue of each group of mice in Experiment 2;

[0025] Figure 10 It is a condition diagram of culturing primary bone marrow macrophages with mouse cardiomyocytes and mouse fibroblasts for 48 hours in Experiment 2;

[0026] Figure 11 It is a diagram of the expression level of the myocardial cell senescence marker (P21) in each group of mice after culture in Experiment 2;

[0027] Figure 12It is a graph showing the expression levels of the fibrotic marker (α-SMA) in the fibroblasts cultured from mice in each group in Experiment 2. Detailed implementation manners

[0028] To make the objectives, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Apparently, the described embodiments are some, but not all, of the embodiments of the present invention. Therefore, the detailed description of the embodiments provided herein is not intended to limit the scope of the claimed invention, but merely represents selected embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the scope of protection of the present invention.

[0029] The present invention has confirmed the application of acteoside in the preparation of drugs for treating senile heart function decline through the following embodiments, but is not limited to these embodiments.

[0030] Experiment 1: Effects of acteoside on improving cardiac function injury and myocardial hypertrophy in aging mice

[0031] 1. Experimental drugs

[0032] The acteoside standard product was purchased from Chengdu Mansite Biotechnology Co., Ltd.

[0033] 2. Experimental animals

[0034] 10 SPF-grade healthy young male C57BL / 6J mice (8 weeks old, body weight 20 ± 3 g), and 30 SPF-grade healthy old male C57BL / 6J mice (80 weeks old, body weight 25 ± 3 g) were all purchased from the Guangdong Provincial Center for Medical Laboratory Animals and raised in the SPF-grade animal experiment center of Guangzhou University of Chinese Medicine. Experimental environment: constant temperature of 20 - 25 °C, relative humidity of 40 - 60%, 12-hour light cycle, free access to food and water. After 1 week of adaptive feeding, the experiment was started.

[0035] 3. Modeling and experimental grouping

[0036] After 1 week of adaptive feeding for 10 young mice, they were used as the control group. After 1 week of adaptive feeding for 30 old mice, they were randomly divided into 3 groups with 10 mice in each group, and the following groups were obtained respectively:

[0037] ① Control group,

[0038] ② Aging group,

[0039] ③ Low-concentration acteoside group (25 mg / kg),

[0040] ④ High-concentration acteoside group (50 mg / kg);

[0041] Calculated according to the body weight of mice, ligustroside was dissolved in 10% DMSO solution + 90% physiological saline to prepare a low-concentration group of ligustroside (25 mg / kg) and a high-concentration group of ligustroside (50 mg / kg), and the elderly mice were gavaged once a day with a gavage volume of 0.2 mL each time; among them, the control group and the senescence group were gavaged with an equal volume of physiological saline for 8 weeks. After the gavage, experiments such as cardiac function detection, cell and tissue staining, real-time fluorescence quantitative PCR, and Western Blot were carried out.

[0042] 4. Data analysis method

[0043] Data are expressed as mean ± standard deviation For comparison among multiple groups, one-way ANOVA was used, and then pairwise comparison was performed using the Turkey test; a P value less than 0.05 was considered statistically significant.

[0044] 5. Experimental results

[0045] As Figures 1 to 3 can be seen, ligustroside has a significant improving effect on myocardial hypertrophy and impaired cardiac function in elderly mice. See Figure 4 , by comparing the heart tissues of the control group (7 weeks old), the senescence group (80 weeks old), and the low-concentration and high-concentration groups (80 weeks old) intervened with ligustroside, we found that the cross-sectional area of myocardial cells in elderly mice was significantly increased compared with that of the young control group (P < 0.05), while ligustroside intervention alleviated myocardial hypertrophy (P < 0.05). See Figure 5 , in the senescence group, the expression levels of inflammatory factors IL-6 and TNF related to the increased risk of cardiovascular events were elevated (P < 0.05), and ligustroside treatment significantly inhibited the inflammatory response (P < 0.05). See Figure 6 , in the hearts of senescent mice, the expression of cardiac function impairment markers BNP, ANP, and NT-proBNP increased (P < 0.05), and the cardiac function was significantly improved after ligustroside intervention (P < 0.05). The above results indicate that ligustroside has a protective effect on the cardiac function of elderly mice and can slow down the impairment of cardiac function during the physiological aging process.

[0046] Experiment 2: Effect of ligustroside on improving myocardial fibrosis and anti-aging after constructing the CHIP model

[0047] 1. Experimental drugs

[0048] The standard product of ligustroside was purchased from Chengdu Mansite Biotechnology Co., Ltd.

[0049] 2. Experimental animals

[0050] 10 SPF - level healthy young male C57BL / 6J mice (8 - week - old, body weight 20 ± 3 g) and 30 SPF - level healthy old male C57BL / 6J mice (80 - week - old, body weight 25 ± 3 g) were all purchased from Guangdong Provincial Medical Laboratory Animal Center and raised in the SPF - level animal experiment center of Guangzhou University of Chinese Medicine. Experimental environment: constant temperature of 20 - 25 °C, relative humidity of 40 - 60%, 12 - hour light cycle, free access to food and water. After 1 - week of adaptive feeding, the experiment began.

[0051] 3. Model establishment and experimental grouping

[0052] After 1 - week of adaptive feeding, 10 young mice were used as the control group. After 1 - week of adaptive feeding of 30 old mice, they were randomly divided into 3 groups with 10 mice in each group, and the following groups were obtained:

[0053] ① Control group,

[0054] ② CHIP model group,

[0055] ③ Low - concentration geniposidic acid group (25 mg / kg),

[0056] ④ High - concentration geniposidic acid group (50 mg / kg);

[0057] CHIP model construction: Old mice were used as recipients for non - myeloablative bone marrow transplantation (BMT). The bone marrow cell suspension of Tet2+ / + or Tet2+ / - mice was transplanted. The bone marrow cells of Tet2+ / - mice (5 × 10 6 cells / day) were continuously injected into recipient mice through the retro - orbital vein for 3 days (a total of 1.5 × 10 7 cells). Subsequently, according to the body weight of the mice, geniposidic acid was dissolved in 10% DMSO solution + 90% physiological saline to prepare the low - concentration geniposidic acid group (25 mg / kg) and the high - concentration geniposidic acid group (50 mg / kg), and they were gavaged once a day with a gavage volume of 0.2 mL each time. Among them, the control group and the CHIP model group were gavaged with an equal volume of physiological saline for 8 weeks. After gavage, cardiac function detection, cell and tissue staining, real - time fluorescence quantitative PCR, and Western Blot and other experiments were carried out.

[0058] 4. Data analysis method

[0059] Data are expressed as mean ± standard deviation For comparison among multiple groups, one - way ANOVA was used, and then pairwise comparison was carried out using Tukey's test; a P - value less than 0.05 was considered statistically significant.

[0060] 5. Experimental results

[0061] (1) By Figures 4 to 6It can be seen that by using genome integration technology, it was found that in patients with cardiovascular diseases, the CHIP signal was specifically elevated and was positively correlated with increasing age. See Figure 4 , through clustering and dimensionality reduction analysis of RNA sequence data (GSE 57338) under different cardiac pathological conditions, individuals carrying and not carrying the CHIP signal were distinguished, and the gene expression profiles of monocytes induced by the CHIP signal in cardiac tissue and peripheral blood were classified. See Figure 5 , the signal mediated by CHIP was significantly enhanced in macrophages ( Figure 5 A) and monocytes ( Figure 5 B), showing significant differences from the non-disease control group. See Figure 6 , the positive correlation between the CHIP signal and increasing age ( Figure 6 A) was demonstrated by a scatter plot. In addition, the mRNA expression levels of the myocardial stress marker (NPPB), the myocardial fibrosis marker (ACTA2), and the inflammation indicators (IL-1β, IL-6) were positively correlated with the upregulation of the CHIP signal ( Figure 6 B - E). Bioinformatics analysis further confirmed that in individuals carrying the CHIP signal, the risk of cardiovascular diseases increased significantly with increasing age.

[0062] (2) It can be seen from Figures 7 to 9 that the CHIP signal is involved in the process of physiological aging, and ligustroside may protect cardiac tissue by slowing down myocardial fibrosis and anti-aging. See Figure 7 , in the cardiac tissue of classic CHIP model mice constructed by competitive bone marrow transplantation Tet2+ / -, the level of the senescence marker P21 increased significantly, where WT indicates transplantation of wild mice and KO indicates transplantation of old mice. See Figure 8 , the results of Sirius red staining showed that compared with the control group, the myocardial fibrosis area in the CHIP model group of mice increased, indicating enhanced fibrotic remodeling in the senescent heart caused by CHIP (P < 0.05). After administration of ligustroside, the generation of myocardial fibrosis was delayed (P < 0.05). See Figure 9 , the qPCR results showed that the transcriptional factor levels of the senescence markers S100A8, S100A9, and SPP1 in the cardiac tissue of the CHIP model group of mice were significantly increased (P < 0.05). After intervention with ligustroside, the transcriptional factor levels of these senescence markers were significantly decreased (P < 0.05).

[0063] (3) See Figure 10, to verify whether ligustroside can slow down the decline of cardiac function by increasing anti-aging levels through inhibiting the CHIP signal, after the CHIP model mice constructed by competitive bone marrow transplantation Tet2+ / - were intervened with ligustroside, bone marrow primary macrophages were sorted by flow cytometry and co-cultured with mouse cardiomyocytes and mouse fibroblasts for 48 hours. As Figures 11 to 12 can be seen, the immunofluorescence results showed that compared with the control group, the levels of the senescence marker (P21) (see Figure 11 ) of cardiomyocytes co-cultured with CHIP model mice and the fibrosis marker (α-SMA) (see Figure 12 ) of fibroblasts were significantly increased (P<0.05), and the levels of these markers were significantly decreased after ligustroside intervention (P<0.05). The results suggest that ligustroside reduces myocardial injury by inhibiting the CHIP signal and increasing anti-aging and fibrosis levels.

[0064] In summary, ligustroside has been proven to be effective in improving myocardial hypertrophy and impaired cardiac function in aged mice, reducing myocardial fibrosis, and decreasing the levels of inflammation and senescence markers, indicating the potential application value of ligustroside in anti-aging and cardiac protection.

[0065] The above description is only the preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and changes. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. Application of Ligustrum lucidum glycosides in the preparation of drugs for treating cardiac dysfunction in the elderly.

2. The use of glossy ligustri lucidi glycoside according to claim 1 in the preparation of a medicine for treating senile heart failure, characterized in that: Application of glossy ligustrum glucoside in the preparation of medicine for treating senile cardiac dysfunction in the aging state.

3. The use of glucosinolate according to claim 2 in the preparation of a medicament for treating senile cardiac dysfunction, characterized in that: The aging state includes any one of the aging states caused by natural decline of body function, coexistence of cardiovascular disease or multiple medications.

4. The use of glossy ligustrum lucidum glycosides according to claim 1 in the preparation of a medicament for treating senile heart failure, characterized in that: The cardiac function impairment includes myocardial fibrosis, cardiac function damage and myocardial hypertrophy.

5. A drug for treating elderly heart failure, characterized in that: The invention comprises ligustri lucidi glycoside and pharmaceutically acceptable auxiliary components.

6. The drug for treating elderly heart failure according to claim 5, characterized in that: The dosage form of the drug includes powder, granules, tablets, capsules, pills, solutions, suspensions or injections.

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