Use of urinary stem cell exosomes in preparation of drugs for preventing and treating atherosclerosis

Abstract

The application provides application of urine-derived stem cell exosomes in preparation of medicines for preventing and treating atherosclerosis, and belongs to the technical field of medicines. The application first finds that the urine-derived stem cell exosomes can improve lipid metabolism, reduce atherosclerotic plaque area and reduce lipid deposition in the liver to play a role in preventing and treating atherosclerosis and fatty liver, and provides a new strategy for preventing and treating atherosclerosis and / or fatty liver.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to the application of urinary stem cell exosomes in the preparation of drugs for the prevention and treatment of atherosclerosis. Background Technology

[0002] According to data provided by the Chinese Center for Disease Control and Prevention in 2017, stroke is the leading cause of death in China. Atherosclerosis is the main cause of vascular diseases such as stroke. Lipid metabolism disorders are the main pathological basis for the development of atherosclerosis. Its characteristic feature is that the lesions in the affected arteries begin in the intima, generally starting with the accumulation of lipids and complex carbohydrates, hemorrhage, and thrombosis, followed by fibrosis and calcification, accompanied by gradual degeneration and calcification of the arterial media, leading to thickening and hardening of the arterial wall and narrowing of the vascular lumen. The lesions often involve large and medium-sized muscular arteries; once they develop to the point of obstructing the arterial lumen, the tissues or organs supplied by that artery will suffer ischemia or necrosis. Currently, drugs for the prevention and treatment of atherosclerosis mainly include lipid-lowering drugs (statins, etc.), antiplatelet drugs (aspirin, etc.), and anticoagulants (warfarin, etc.). However, these drugs all have certain side effects, which affect long-term use and patients' confidence in treatment. Therefore, providing a safe and effective drug for the prevention and treatment of atherosclerosis is particularly important.

[0003] Urinary stem cells are a type of stem cell with multi-lineage differentiation and self-renewal capabilities. Due to their numerous advantages, such as easy access, abundant sources, simple culture, and lack of ethical concerns, they are widely studied and applied. However, direct stem cell transplantation always carries certain risks, such as vascular embolism, microinfarction, and tumor formation. Therefore, it is necessary to explore new strategies that can fully utilize the superior functions of stem cells while avoiding the potential risks of stem cell transplantation. Recent studies have found that only a very small percentage of transplanted stem cells directly differentiate into specific parenchymal cells; they primarily mobilize endogenous cells to participate in tissue repair and regeneration through paracrine secretion of bioactive molecules. Exosomes are membrane-bound vesicles containing various RNAs and proteins secreted by eukaryotic cells, which can facilitate information transmission between the source cell and surrounding cells through paracrine secretion. Existing technologies disclose the use of urinary stem cell exosomes for the treatment of osteoporosis, but there are no reports of their application in the prevention and treatment of atherosclerosis. Summary of the Invention

[0004] This invention provides the application of urinary stem cell exosomes in the preparation of drugs for the prevention and treatment of atherosclerosis, with the aim of solving the aforementioned problems existing in the background art.

[0005] To achieve the above objectives, embodiments of the present invention provide the application of urinary stem cell exosomes in the preparation of drugs for the prevention and treatment of atherosclerosis.

[0006] Furthermore, the urinary stem cell exosomes are used to regulate blood lipids.

[0007] Furthermore, the urinary stem cell exosomes are used to reduce the area of ​​atherosclerotic plaques.

[0008] Furthermore, the urinary stem cell exosomes are used to reduce pathological lipid deposition in the liver.

[0009] Furthermore, the drug also includes a pharmaceutically acceptable carrier, which is at least one of excipients, fillers, binders, wetting agents, sustained-release agents, absorption enhancers, surfactants, and lubricants.

[0010] Furthermore, the drug dosage form is a tablet, pill, capsule, or injection.

[0011] Furthermore, the drug may also be subject to any form of functional enhancement modification, which refers to linking the urinary stem cell exosomes to a targeted aptamer / antibody, or encapsulating them with nanoparticles.

[0012] Furthermore, the subjects of the drug had, were suspected of having, or suffered from atherosclerosis and / or lipid metabolism-related diseases.

[0013] Furthermore, the subjects are humans or non-human mammals.

[0014] This invention demonstrates through animal experiments that urinary stem cell exosomes are highly effective in improving atherosclerosis in ApoE knockout mice fed a high-fat diet and New Zealand white rabbits with balloon-induced aortic injury. In the ApoE mouse atherosclerosis model fed a high-fat diet, after intervention with urinary stem cell exosomes, gross Oil Red O staining results showed that the area of ​​atherosclerotic plaques in the arteries was significantly reduced compared with the solvent control group (P < 0.05); serological tests showed that the serum triglyceride content was significantly reduced compared with the solvent control group (P < 0.05). In a New Zealand white rabbit model of atherosclerosis induced by balloon-induced aortic injury after high-fat feeding, H&E staining, Oil Red O staining, Masson staining, and α-SMA staining showed that the area of ​​atherosclerotic plaques in the arteries was significantly reduced compared to the solvent control group (P < 0.05), while the collagen content and stability of the plaques were significantly increased compared to the solvent control group (P < 0.05). Serological tests showed that the serum triglyceride content was significantly reduced compared to the solvent control group (P < 0.05).

[0015] Therefore, urinary stem cell exosomes can be used to prepare drugs and drug compositions for the prevention and treatment of atherosclerosis, providing a new, safe and effective biological treatment method for the prevention and treatment of atherosclerosis.

[0016] The above-described solution of the present invention has the following beneficial effects:

[0017] This invention provides the application of urinary stem cell exosomes in the preparation of drugs for the prevention and treatment of atherosclerosis. By intravenously injecting urinary stem cell exosomes extracted from the body, blood lipids are reduced and the area of ​​atherosclerotic plaques is decreased, thereby achieving the purpose of preventing and treating atherosclerosis. This provides a new strategy for the safe and effective prevention and treatment of atherosclerosis. Attached Figure Description

[0018] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 This is a transmission electron microscope image of exosomes derived from urinary stem cells in an embodiment of the present invention;

[0020] Figure 2 This is a Western blot image of exosome surface markers from urinary stem cells according to an embodiment of the present invention.

[0021] Figure 3 This is a comparison of the results of Oil Red O staining of the aorta in ApoE knockout mice after three months of treatment with normal or high-fat diets and intervention with phosphate solution or urinary stem cell exosomes in Example 1 of this invention. Figure 3 A refers to a representative image of the gross Oil Red O staining of the aorta. Figure 3 B refers to the statistical data of gross Oil Red O staining;

[0022] Figure 4 These are the test results of blood lipid levels in each group in Example 1 of this invention;

[0023] Figure 5 This is a comparison of the H&E staining, Oil Red O staining, Masson staining, and α-SMA staining results of a rabbit atherosclerosis model with balloon-injured aorta after three months of intervention with phosphate solution or urinary stem cell exosomes under normal or high-fat diet feeding conditions. Figure 5 A refers to representative images and statistical data of H&E staining of the abdominal aorta of each group of rabbits. Figure 5 B refers to representative images and statistical data of Oil Red O staining of the abdominal aorta of each group of rabbits. Figure 5 C refers to representative images and statistical data of Masson staining of the abdominal aorta of each group of rabbits. Figure 5 D refers to representative images and statistical data of immunohistochemical staining of α-SMA in the abdominal aorta of each group of rabbits;

[0024] Figure 6 These are the test results of blood lipid levels in each group in Example 2 of this invention. Detailed Implementation

[0025] To make the technical problems, technical solutions and advantages of the present invention clearer, a detailed description will be given below in conjunction with the accompanying drawings and specific embodiments.

[0026] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those skilled in the art. The technical terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the scope of the invention.

[0027] Unless otherwise specified, all raw materials, reagents, instruments and equipment used in this invention can be purchased from the market or prepared by existing methods.

[0028] This invention addresses existing problems by providing the application of urinary stem cell exosomes in the preparation of drugs for the prevention and treatment of atherosclerosis.

[0029] Furthermore, the urinary stem cell exosomes are used to regulate blood lipids.

[0030] Furthermore, the urinary stem cell exosomes are used to reduce the area of ​​atherosclerotic plaques.

[0031] Furthermore, the urinary stem cell exosomes are used to reduce pathological lipid deposition in the liver.

[0032] Furthermore, the drug also includes a pharmaceutically acceptable carrier, which is at least one of excipients, fillers, binders, wetting agents, sustained-release agents, absorption enhancers, surfactants, and lubricants.

[0033] Furthermore, the drug dosage form is a tablet, pill, capsule, or injection.

[0034] Furthermore, the drug may also be subject to any form of functional enhancement modification, which refers to linking the human urinary stem cell exosomes to a targeted aptamer / antibody, or encapsulating them with nanoparticles.

[0035] Furthermore, the subjects of the drug had, were suspected of having, or suffered from atherosclerosis and / or lipid metabolism-related diseases.

[0036] Furthermore, the subjects are humans or non-human mammals.

[0037] The inventors of this invention conducted animal experiments using urinary stem cell exosomes. By intervening in ApoE knockout mice fed a high-fat diet and New Zealand rabbits with balloon injury fed a high-fat diet, they observed that urinary stem cell exosomes could significantly inhibit the formation of atherosclerotic plaques.

[0038] Figure 1 Transmission electron microscopy image of exosomes from urinary stem cells;

[0039] Figure 2 This is a Western blot image of exosome surface markers from urinary stem cells.

[0040] Example 1

[0041] Urinary stem cell exosomes can prevent and treat atherosclerotic plaque formation in ApoE knockout mice fed a high-fat diet.

[0042] Mouse model: Six-week-old ApoE knockout male SPF-grade mice, weighing 18–20 g, were purchased from Cavens Laboratory Animal Co., Ltd., Changzhou, Jiangsu Province. Each group consisted of 7–10 mice, with free access to food and water, and a circadian rhythm was maintained.

[0043] Grouping experiment: ApoE knockout male SPF mice were randomly divided into 3 groups, including normal diet + solvent control group, high-fat diet + solvent control group, and high-fat diet + urinary stem cell exosome group; each group consisted of 7 to 10 mice.

[0044] Modeling: Mice in each group were fed different diets while undergoing intervention.

[0045] Administration method:

[0046] Normal diet + solvent control group (ND) M Mice were fed a normal diet and simultaneously received phosphate-buffered saline injections via the tail vein at a volume of 100 μL, once a week. Samples were collected after three months of intervention.

[0047] High-fat diet + solvent control group (HFD) M Mice were fed a high-fat diet and received phosphate-buffered saline via tail vein injection at a volume of 100 μL once a week. Samples were collected after three months of intervention.

[0048] High-fat diet + urinary stem cell exosome group (HFD) M +USC-Exos): Mice were fed a high-fat diet and simultaneously injected via tail vein into urinary stem cell exosomes at a volume of 100 μL once a week. Samples were collected after three months of intervention.

[0049] like Figure 3The image shows a comparison of Oil Red O staining results of the aorta in ApoE knockout mice after three months of treatment with normal or high-fat diets and intervention with phosphate solution or urinary stem cell exosomes. Note: *P<0.05 indicates that the difference was statistically significant compared with the negative control group (ApoE knockout mice fed a normal diet). # P<0.05 indicates that the difference was statistically significant compared with the solvent control group (ApoE knockout mice fed a high-fat diet); Figure 3 A refers to a representative image of the gross Oil Red O staining of the aorta. Figure 3 B refers to the statistical data of gross Oil Red O staining;

[0050] like Figure 4 The results of blood lipid levels in each group are shown in the figure; Note: *P<0.05 indicates that the difference is statistically significant compared with the negative control group. # P<0.05 indicates that the difference is statistically significant compared with the solvent control group;

[0051] Example 2

[0052] Urinary stem cell exosomes can prevent atherosclerotic plaque formation in the aorta of New Zealand rabbits with high-fat diet and balloon-induced aortic injury.

[0053] Rabbit model: Ten-week-old New Zealand white rabbits, weighing 2.5–3.0 kg, were selected and purchased from Hunan Slack Jingda Animal Experiment Co., Ltd. Each group consisted of 5–7 New Zealand white rabbits, with free access to food and water, and their diurnal rhythm was maintained.

[0054] Grouping experiment: New Zealand white rabbits were randomly divided into 3 groups, including normal diet + solvent control group, high-fat diet + solvent control group, and high-fat diet + urinary stem cell exosome group; each group had 5 to 7 rabbits.

[0055] Modeling: After feeding the animals with normal or high-fat diets for two weeks, balloon injury was performed on each group. After balloon injury, each group was treated with phosphate buffer or urinary stem cell exosome solution.

[0056] Administration method:

[0057] Normal feed + solvent control group (ND) R Following balloon injury, New Zealand rabbits were fed a normal diet while receiving phosphate-buffered saline via ear vein injection at a volume of 2 mL, once a week. Samples were collected three months after the intervention.

[0058] High-fat diet + solvent control group (HFD) R Following balloon injury, New Zealand rabbits were fed a high-fat diet and simultaneously given phosphate-buffered saline via ear vein injection at a volume of 2 mL, once a week. Samples were collected three months after the intervention.

[0059] High-fat diet + urinary stem cell exosome group (HFD) R +USC-Exos): After balloon injury, New Zealand rabbits were fed a normal diet and simultaneously injected with urinary stem cell exosome solution via ear vein. The volume of the ear vein injection was 2 mL, once a week. Samples were collected after three months of intervention.

[0060] like Figure 5 The image shows a comparison of H&E staining, Oil Red O staining, Masson staining, and α-SMA staining results of vascular sections in a rabbit atherosclerosis model with balloon-injured aorta after three months of intervention with phosphate solution or urinary stem cell exosomes, under normal or high-fat diet conditions. Note: *P<0.05 indicates a statistically significant difference compared to the negative control group (New Zealand white rabbits with balloon-injured aorta fed a normal diet). # P<0.05 indicates that the difference was statistically significant compared with the solvent control group (New Zealand white rabbits fed a high-fat diet with balloon-injured aorta). Figure 5 A refers to representative images and statistical data of H&E staining of the abdominal aorta of each group of rabbits. Figure 5 B refers to representative images and statistical data of Oil Red O staining of the abdominal aorta of each group of rabbits. Figure 5 C refers to representative images and statistical data of Masson staining of the abdominal aorta of each group of rabbits. Figure 5 D refers to representative images and statistical data of immunohistochemical staining of α-SMA in the abdominal aorta of each group of rabbits;

[0061] like Figure 6 The results of blood lipid levels in each group are shown in the figure; Note: *P<0.05 indicates that the difference is statistically significant compared with the negative control group. # P<0.05 indicates that the difference is statistically significant compared with the solvent control group;

[0062] The above description represents the preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. The use of urinary stem cell exosomes in the preparation of a medicament for preventing and treating atherosclerosis, characterized in that, The urine-derived stem cell exosome is used for reducing the area of atherosclerotic plaque.

2. Use according to claim 1, characterized in that, The urine-derived stem cell exosome is used for regulating blood lipid.

3. Use according to claim 1, characterized in that, The urine-derived stem cell exosome is used for reducing pathological lipid deposition in the liver.

4. Use according to claim 1, characterized in that, The medicine further comprises a pharmaceutically acceptable carrier, which is at least one of an excipient, a filler, a binder, a humectant, a sustained-release agent, an absorption enhancer, a surfactant, a lubricant.

5. The use according to claim 1, characterized in that, The medicine dosage form is a tablet, a pill, a capsule or a needle.

6. Use according to claim 1, characterized in that, The medicine can also be functionally enhanced by modification, which means that the urine-derived stem cell exosome is connected with an aptamer / antibody with targeting property or wrapped with a nanoparticle.

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