Application of LS-102 in preparation of medicine for preventing and treating myocardial ischemia-reperfusion injury

Abstract

The invention discloses application of LS-102 in preparation of a medicine for treating myocardial ischemia-reperfusion injury, and belongs to the technical field of biological medicine. A contrast or LS-102 treated mouse is taken as an experimental subject, a myocardial ischemia reperfusion injury model is formed by ligating the left anterior descending coronary artery (LAD) of the heart of the mouse, and the result shows that compared with the contrast mouse, the LS-102 treated mouse has the advantages that the cardiac infarction area is reduced, the myocardial injury marker level is reduced, the cardiac microcirculation perfusion is increased, and the inflammatory cell infiltration is reduced. The application of the LS-102 in relieving the myocardial ischemia reperfusion injury is proved for the first time, a novel medicine molecule is provided for clinically treating the myocardial ischemia reperfusion injury, and the application prospect and clinical value are good.

Description

Technical Field

[0001] The invention relates to application of LS-102 in preparing medicine for preventing and treating myocardial ischemia-reperfusion injury, and belongs to the technical field of biomedicine. Background Art

[0002] Coronary heart disease and acute myocardial infarction are the most important critical diseases threatening human health. At present, the incidence and mortality of acute myocardial infarction in my country are still on the rise. The only way to save ischemic myocardium is timely reperfusion therapy, including drug thrombolysis and percutaneous coronary intervention. However, reperfusion therapy also induces a special irreversible injury, called ischemia-reperfusion injury. It is closely related to clinical complications such as severe arrhythmias, heart failure, and sudden death. Therefore, in-depth research on the specific pathological mechanism of myocardial ischemia-reperfusion injury and exploration of potential myocardial therapeutic targets are of great clinical translational significance.

[0003] Astragalus is a traditional Chinese medicine ingredient that plays an important role in cardiovascular disease and immune regulation. Huangshi formic acid (LS-102) is a new derivative of astragaloside IV, which has antioxidant, anti-inflammatory and anti-apoptotic effects. In addition, LS-102 is also a selective E3 ligase synoviolin (Syvn1) inhibitor. However, the role of LS-102 in myocardial ischemia-reperfusion injury has not been reported. Summary of the invention

[0004] The purpose of the present invention is to provide the use of LS-102 in the preparation of a drug for preventing and treating myocardial ischemia-reperfusion injury. The results of the present invention show that LS-102 can reduce myocardial ischemia-reperfusion injury and can be further used to improve the prognosis of patients with myocardial infarction.

[0005] In order to achieve the above object, the present invention provides the use of LS-102, the chemical structure of LS-102 is as follows:

[0006]

[0007] The use is selected from any one of the following 1)-5):

[0008] 1) Preparation of drugs for preventing myocardial ischemia-reperfusion injury;

[0009] 2) Preparation of drugs for treating myocardial ischemia-reperfusion injury;

[0010] 3) Preparation of drugs for preventing myocardial ischemia-reperfusion injury from developing into acute myocardial infarction;

[0011] 4) Preparation of drugs for treating acute myocardial infarction caused by myocardial ischemia-reperfusion injury;

[0012] 5) Prepare drugs for improving the prognosis of myocardial infarction.

[0013] Preferably, the drug comprises an active ingredient and a pharmaceutically acceptable excipient, and the active ingredient is LS-102.

[0014] Preferably, the pharmaceutically acceptable excipient is selected from one or more combinations of buffers, encapsulating agents, fillers, adhesives, transdermal absorbents, wetting agents, disintegrants, absorption enhancers, surfactants, colorants, flavoring agents and adsorption carriers.

[0015] Preferably, the dosage form of the drug is an injection or an oral preparation.

[0016] Preferably, when the LS-102 is used to prevent and treat the myocardial ischemia-reperfusion injury model, its usage and dosage are as follows: LS-102 (4 mg / kg) is injected intraperitoneally into mice 12 hours before coronary artery ligation and immediately after surgery.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] The experimental results of the present invention show that in myocardial ischemia-reperfusion injury caused by coronary artery ligation treated with LS-102, the myocardial infarction area of ​​mice is significantly reduced, the level of myocardial injury markers is reduced, the cardiac microcirculation perfusion is significantly increased, and the inflammatory cell infiltration is reduced; it is proved that LS-102 can improve myocardial ischemia-reperfusion injury, provide a new drug target for the treatment of myocardial ischemia-reperfusion injury, and has very important clinical transformation value. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The results of Evans blue / TTC staining show that LS-102 reduces the myocardial infarction area in mice induced by myocardial ischemia-reperfusion injury;

[0020] Figure 2 The results of serum LDH and CK-MB, markers of myocardial injury in mice, were shown to be LS-102-induced myocardial ischemia-reperfusion injury;

[0021] Figure 3 To use LS-102 to alleviate the microcirculatory perfusion disorder induced by myocardial ischemia-reperfusion injury in mice;

[0022] Figure 4 The purpose of this study is to use LS-102 to reduce cardiac inflammatory cell infiltration induced by myocardial ischemia-reperfusion injury. DETAILED DESCRIPTION

[0023] In order to make the present invention more clearly understood, preferred embodiments are described in detail below with reference to the accompanying drawings.

[0024] Example

[0025] This example provides the use of LS-102 in the preparation of a drug for preventing and treating myocardial ischemia-reperfusion injury:

[0026] 1. Experimental methods

[0027] 1.1 Establishment of myocardial ischemia-reperfusion injury model in mice. The specific steps include:

[0028] (1) Before surgery, mice were anesthetized with 2% isoflurane and fixed on a constant temperature operating table;

[0029] (2) Depilation of the mouse chest area and disinfection with 75% alcohol;

[0030] (3) Make a small incision of about 0.5 cm along the left edge of the sternum and use hemostatic forceps to bluntly separate the muscle layer;

[0031] (4) Enter the chest cavity, pierce the pericardium, and gently squeeze the mouse chest cavity with the left hand to fully expose the heart outside the chest wall. Use 6-0 surgical sutures to ligate the left anterior descending coronary artery below the edge of the left atrial appendage to create a live knot;

[0032] (5) Once completed, the heart is returned to the chest cavity, the air is squeezed out, and the incision is sutured;

[0033] (6) After 45 minutes of ischemia, the joint was released, coronary blood flow was restored, and the wound was closed with 4-0 surgical sutures;

[0034] (7) Successful reperfusion was defined as an ST segment decrease of more than 50% 1 hour after the knot was loosened on the electrocardiogram.

[0035] 1.2 Cardiac blood perfusion detection

[0036] The mouse was fixed on the operating table. The tail was wiped with alcohol. After the location of the mouse tail vein was determined, Lectin (10 μg / mL) was injected into the mouse tail vein with an insulin needle. After 5 minutes, the mouse was anesthetized and killed, and the heart was immediately perfused with 20 mL PBS to remove the blood containing Lectin. The heart was removed, trimmed, and placed on absorbent paper to remove moisture. The heart was placed in an OCT embedding box in a fixed direction to avoid bubbles, and the basic information of the sample was marked on the box. The OCT embedding box containing the sample was slowly immersed in the liquid nitrogen surface and slowly frozen. After the embedding agent completely turned white and solidified, it was completely immersed in liquid nitrogen. The heart tissue was frozen and sectioned with a thickness of 7 μm. 150 μL of fixative was added to the tissue and fixed at room temperature for 30 minutes. The fixative was aspirated and washed three times with punching solution at room temperature for 5 minutes each time, and then the punching solution was added again and incubated at room temperature for 15 minutes. The slide was sealed with DAPI mounting medium to ensure that the coverslip was tight to the tissue patch and there were no bubbles. Confocal microscope was used for photography.

[0037] 1.3 Flow cytometry sorting of cardiac inflammatory cells

[0038] Prepare cardiac tissue digestion enzymes including collagenase II (1 mg / mL), elastase (0.05 mg / mL) and DNA enzyme (0.05 mg / mL), and prepare them with 1640 culture medium according to their respective proportions to make cardiac tissue digestion solution. First, add the cardiac tissue to 100 μL digestion solution, cut the tissue into pieces, add 5 mL digestion solution, place it in a 15 mL centrifuge tube, digest it in a 37°C water bath for 40 minutes, and then put the sample on ice. Gently blow the sample with a 1 mL pipette and filter it with a 70 μm filter. Add 2 mL of red blood cell lysis solution to the cardiac tissue cell pellet, blow it gently, lyse it at room temperature for 5 minutes, then centrifuge it for 5 minutes, and discard the supernatant. After washing with PBS, flow staining was performed, and inflammatory cells (CD11b + Ly6C + ). Add 200 μL of antibody to the heart tissue cell pellet from the previous step and blow evenly. Incubate at 4°C in the dark for 40 minutes, and set up isotype control tubes and single-staining tubes at the same time. After washing, resuspend the heart tissue cell pellet and use FACSAria flow cytometer to sort the stained heart samples.

[0039] To verify the protective effect of LS-102 treatment on the mouse myocardium, ischemia-reperfusion injury models of mice were established with control treatment and LS-102 treatment, respectively. After 24 hours of perfusion, Evans blue / TTC staining was used to detect the myocardial infarction area, serum LDH and CK-MB were used to detect myocardial injury, Lectin was used to detect cardiac blood perfusion, and flow cytometry was used to detect cardiac inflammatory cell infiltration. The results showed that LS-102 treatment significantly reduced the myocardial infarction area, improved blood perfusion and inhibited inflammatory cell infiltration.

[0040] 2. Experimental results:

[0041] (1) Evans blue / TTC staining results showed that LS-102 treatment significantly reduced the myocardial infarction area induced by myocardial ischemia-reperfusion injury in mice. Figure 1 shown.

[0042] (2) The results of serum myocardial injury marker detection showed that LS-102 treatment alleviated the increase in serum LDH and CK-MB levels in mice induced by myocardial ischemia-reperfusion injury. Figure 2 shown.

[0043] (3) The results of cardiac blood perfusion test showed that LS-102 treatment significantly alleviated the blood perfusion disorder induced by myocardial ischemia-reperfusion injury, such as Figure 3 shown.

[0044] (4) Flow cytometry showed that LS-102 treatment significantly reduced the infiltration of inflammatory cells induced by myocardial ischemia-reperfusion injury. Figure 4 shown.

[0045] The above description is only a preferred embodiment of the present invention and is not any formal or substantial limitation of the present invention. It should be pointed out that a person skilled in the art can make several improvements and supplements without departing from the present invention, and these improvements and supplements should also be regarded as the protection scope of the present invention.

Claims

1. The use of LS-102 is characterized by: The chemical structural formula of LS-102 is as follows: The use is selected from any one of the following 1)-5): 1) Preparation of drugs for preventing myocardial ischemia-reperfusion injury; 2) Preparation of drugs for treating myocardial ischemia-reperfusion injury; 3) Preparation of drugs for preventing myocardial ischemia-reperfusion injury from developing into acute myocardial infarction; 4) Preparation of drugs for treating acute myocardial infarction caused by myocardial ischemia-reperfusion injury; 5) Prepare drugs for improving the prognosis of myocardial infarction.

2. The use according to claim 1, characterized in that The medicine comprises an active ingredient and pharmaceutically acceptable excipients, and the active ingredient is LS-102.

3. The use according to claim 2, characterized in that The pharmaceutically acceptable excipients are selected from one or more combinations of buffers, encapsulating agents, fillers, adhesives, transdermal absorbents, wetting agents, disintegrants, absorption promoters, surfactants, colorants, flavoring agents and adsorption carriers.

4. The use according to claim 2, characterized in that The dosage form of the medicine is injection or oral preparation.

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