Application of kaempferol-3-O-neohesperidin in preparation of medicine for preventing and treating myocardial injury

Abstract

The invention provides a new application of a small molecule compound kaempferol-3-O-neohesperidin in pollen typhae in preparation of a medicine for preventing and / or treating myocardial injury. It is found for the first time that kaempferol-3-O-neohesperidin can reduce myocardial damage by inhibiting myocardial cell death and increasing the ATP content in myocardial cells, and can be used for preparing the medicine for preventing and treating myocardial damage. Specifically, the kaempferol-3-O-neohesperidin can relieve myocardial cell oxidative damage caused by H2O2, and has a myocardial protection effect. The invention not only contributes to the development and utilization of the pollen typhae, but also has important significance on the research and development of medicines for preventing and / or treating myocardial injury.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine and relates to a small molecule kaempferol-3-O-neohesperidin, and specifically relates to the application of kaempferol-3-O-neohesperidin in the preparation of a drug for preventing and treating myocardial injury. Background Art

[0002] Cardiovascular disease (CVD) refers to circulatory system diseases including hypertension, coronary heart disease, heart failure and arrhythmia, which are characterized by high morbidity and mortality and seriously threaten human health. According to data from the World Health Organization (WHO), nearly 17.9 million people die from CVD each year, accounting for 31% of the total number of deaths. During the development of CVD, lesions in the heart and blood vessels can easily lead to myocardial ischemia and hypoxia, and even necrosis, which in turn leads to cardiac dysfunction. Therefore, the repair and treatment of myocardial damage is the core of CVD research. Studies have shown that traditional Chinese medicine can reduce myocardial damage by inhibiting ferroptosis, apoptosis, oxidative stress, regulating autophagy and regulating energy metabolism. At present, the treatment of CVD mainly includes lifestyle improvement, drug therapy and surgical intervention. Although clinical treatment has made certain progress, CVD patients still face many challenges during the treatment process, such as large drug side effects, high treatment costs and long treatment cycles. Therefore, there is an urgent need to seek safer, more effective and economical treatment options.

[0003] Typhae pollen is the dried pollen of Typha truncatum and Typha orientalis, plants of the Typhaceae family. It has the effects of cooling blood and stopping bleeding, promoting blood circulation and removing blood stasis. It is commonly used in the clinic to treat various bleeding syndromes and stasis pain syndromes, and has the effects of improving myocardial microcirculation, lowering blood lipids, resisting atherosclerosis, and promoting blood coagulation. Modern pharmacological studies have shown that Typhae pollen has the effects of analgesia, anti-inflammatory, anti-hypobaric hypoxia, promoting blood circulation, lowering blood lipids, preventing arteriosclerosis, and enhancing immunity, and has good application prospects. Therefore, the separation and identification of active small molecule compounds in Typhae pollen is of great significance in this field. Summary of the invention

[0004] In view of the deficiencies of the prior art, the object of the present invention is to provide an application of kaempferol-3-O-neohesperidin in the preparation of a drug for preventing and treating myocardial injury.

[0005] In a first aspect of the present invention, there is provided a use of kaempferol-3-O-neohesperidin in the preparation of a drug for improving the survival rate of cardiomyocytes, wherein the structural formula of the kaempferol-3-O-neohesperidin is:

[0006]

[0007] Furthermore, the cardiomyocytes are oxidatively damaged cardiomyocytes.

[0008] Furthermore, the oxidative damage is oxidative damage caused by H2O2.

[0009] In the second aspect of the present invention, there is provided a use of kaempferol-3-O-neohesperidin in the preparation of a drug for improving the ATP level of cardiomyocytes, wherein the structural formula of the kaempferol-3-O-neohesperidin is:

[0010]

[0011] Furthermore, the cardiomyocytes are oxidatively damaged cardiomyocytes.

[0012] Furthermore, the oxidative damage is oxidative damage caused by H2O2.

[0013] In the third aspect of the present invention, there is provided the use of kaempferol-3-O-neohesperidin in the preparation of a myocardial protective drug, characterized in that the structural formula of the kaempferol-3-O-neohesperidin is:

[0014]

[0015] In a fourth aspect of the present invention, there is provided a use of kaempferol-3-O-neohesperidin in the preparation of a medicament for preventing and / or treating myocardial injury, wherein the structural formula of the kaempferol-3-O-neohesperidin is:

[0016]

[0017] Furthermore, the myocardial damage is oxidative damage.

[0018] Furthermore, the oxidative damage is oxidative damage caused by H2O2.

[0019] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

[0020] Compared with the prior art, the present invention has the following advantages and progress:

[0021] The present invention provides a new use of kaempferol-3-O-neohesperidin, a small molecule compound in Pollen Typhae, in the preparation of a drug for preventing and / or treating myocardial injury. The present invention first discovered that kaempferol-3-O-neohesperidin can reduce myocardial injury by reducing myocardial cell death and increasing the ATP content in myocardial cells, and can be used to prepare a drug for preventing and treating myocardial injury. Specifically, kaempferol-3-O-neohesperidin can reduce oxidative damage to myocardial cells caused by H2O2 and has a myocardial protective effect. The present invention is not only conducive to the development and utilization of Pollen Typhae, but also has important significance for the research and development of drugs for preventing and / or treating myocardial injury. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] The accompanying drawings, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0023] Figure 1 : The chemical structural formula of kaempferol-3-O-neohesperidin.

[0024] Figure 2 : Effect of kaempferol-3-O-neohesperidin treatment on the survival rate of H9c2 cardiomyocytes injured by H2O2 (compared with the control group, # P<0.05; compared with the H2O2 group, * P<0.05, ** P<0.01, n=4).

[0025] Figure 3 :Effects of kaempferol-3-O-neohesperidin on ATP content in H9c2 cardiomyocytes injured by H2O2 (compared with the control group, ## P<0.01; compared with the H2O2 group, ** P<0.01, n=3). DETAILED DESCRIPTION

[0026] The present invention provides the use of kaempferol-3-O-neohesperidin in the preparation of drugs for preventing and treating myocardial injury. The present invention is specifically described below in conjunction with the embodiments to facilitate further understanding of the present invention by those skilled in the art. However, the embodiments described below are only part of the embodiments of the present invention and should not be regarded as any form of limitation to the present invention. It should be pointed out that adjustments and improvements made by ordinary technicians in this field based on the concept of the present invention should be regarded as the scope of protection of the present invention. The specific technical operation steps and operators are not indicated in the embodiments, and they are all carried out in accordance with the general technical conditions described in the literature in this field or the relevant product instructions.

[0027] Example

[0028] Part I Materials and Methods

[0029] 1 Materials

[0030] 1.1 Drugs and reagents

[0031] Kaempferol-3-O-neohesperidoside (CAS No.: 32602-81-6, chemical structure as Figure 1 The above samples were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; hydrogen peroxide (H2O2) was purchased from Sigma; penicillin / streptomycin and fetal bovine serum were purchased from Gibco; BCA protein detection kit was purchased from Solebao Biotechnology Co., Ltd.; ATP detection kit was purchased from Shanghai Bio-Tech Biotechnology Co., Ltd.

[0032] 1.2 Experimental cells

[0033] Rat H9c2 cardiomyocytes were purchased from ATCC, USA.

[0034] 1.3 Experimental instruments

[0035] HF240 carbon dioxide incubator (China Likang Biomedical Technology Holdings Co., Ltd.); clean bench (Shanghai Yuejin Medical Instrument Co., Ltd.); microscope (OLYMPUS Corporation); The microplate reader was used (Tecan, Switzerland).

[0036] 2 Methods

[0037] 2.1 Cell culture and grouping

[0038] H9c2 cells were routinely cultured in a constant temperature incubator at 37°C and 5% CO2 using DMEM high-glucose medium containing 10% fetal bovine serum and 1% penicillin / streptomycin. When the cells grew to 80% to 90%, they were passaged and cells in the logarithmic growth phase were used for subsequent experiments. The cells were divided into a normal group, a model group (H2O2, 300μmol / L), and groups treated with different concentrations of kaempferol-3-O-neohesperidin. The cells in the normal group were not treated. The model group was treated with 300μmol / L H2O2 for 2h. The kaempferol-3-O-neohesperidin group was first treated with different concentrations of drugs for 24h, and then co-treated with kaempferol-3-O-neohesperidin and H2O2 for 2h.

[0039] 2.2 Cell viability determination

[0040] The cells (5×10 3 After the treatment with drugs and H2O2, 100 μL / well serum-free medium was replaced, 10 μL / well CCK-8 stock solution was added, and the cells were incubated in a 5% CO2 incubator for 1 to 2 h. The absorbance was measured at 450 nm.

[0041] 2.3 ATP determination

[0042] H9c2 cells in the logarithmic growth phase were taken at 1.5×10 5 Each well was inoculated in a 6-well plate. After treatment with drugs and H2O2, cells in each group were collected and tested according to the instructions of the ATP kit.

[0043] 3 Statistical analysis

[0044] GraphPad Prism 8.0 software was used to analyze the experimental data, and the data are expressed as mean ± standard error. The data were tested for normality. If they were normally distributed, the Student's t test was used for comparison between the two groups, and one-way ANOVA was used for comparison between multiple groups. If they did not meet the normality test, the nonparametric test (Kruskal-Wallis rank sum test) was used. P < 0.05 was considered statistically significant.

[0045] Part II Experimental Results

[0046] Effect of kaempferol-3-O-neohesperidin on the survival rate of myocardial cells injured by H2O2

[0047] Effects of compounds on survival rate of H9c2 cardiomyocytes induced by H2O2 Figure 2 Compared with the normal group, the cell viability of the model group was significantly decreased (P<0.05); compared with the model group, the cell viability of the 50 and 100 μM kaempferol-3-O-neohesperidin treatment groups was significantly increased (P<0.05, P<0.01), indicating that 50 and 100 μM kaempferol-3-O-neohesperidin can protect the viability of cardiomyocytes. The above results show that kaempferol-3-O-neohesperidin can significantly improve the survival rate of cardiomyocytes damaged by H2O2, indicating that the compound has cardioprotective activity.

[0048] Effects of 2-kaempferol-3-O-neohesperidin on ATP in H2O2-damaged cardiomyocytes

[0049] Effects of compounds on ATP in H2O2-damaged myocardial cells Figure 3 Compared with the blank group, the ATP content in the model group was significantly decreased (P<0.01); compared with the model group, the ATP content was significantly increased after treatment with different concentrations of kaempferol-3-O-neohesperidin (P<0.01), indicating that kaempferol-3-O-neohesperidin can increase the ATP level of myocardial cells injured by H2O2.

[0050] Conclusion: The above results show that kaempferol-3-O-neohesperidin can reduce the oxidative damage of myocardial cells induced by H2O2 and has a cardioprotective effect.

[0051] The above is only a preferred embodiment of the present invention and does not limit the present invention in any way. For those skilled in the art, any modification and change made to the above embodiment according to the technical essence of the present invention shall fall within the protection scope of the technical solution of the present invention.

Claims

1. Use of kaempferol-3-O-neohesperidin in the preparation of a drug for improving the survival rate of cardiomyocytes, characterized in that: The structural formula of the kaempferol-3-O-neohesperidin is:

2. The use according to claim 1, characterized in that: The cardiomyocytes are oxidatively damaged cardiomyocytes.

3. Use of kaempferol-3-O-neohesperidin in the preparation of a drug for improving the ATP level of myocardial cells, characterized in that: The structural formula of the kaempferol-3-O-neohesperidin is:

4. The use according to claim 3, characterized in that: The cardiomyocytes are oxidatively damaged cardiomyocytes.

5. Use of kaempferol-3-O-neohesperidin in the preparation of a myocardial protective drug, characterized in that: The structural formula of the kaempferol-3-O-neohesperidin is:

6. Use of kaempferol-3-O-neohesperidin in the preparation of a drug for preventing and / or treating myocardial injury, characterized in that: The structural formula of the kaempferol-3-O-neohesperidin is:

7. The use according to claim 6, characterized in that: The myocardial damage is oxidative damage.

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