New application of aescin
By using aescin in drug preparation, AMPK is activated, mTOR phosphorylation is downregulated, and autophagy is induced. Aescin can effectively reduce cholesterol accumulation in foam cells and improve atherosclerosis.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- WUXI NO 2 PEOPLES HOSPITAL
- Filing Date
- 2023-04-17
- Publication Date
- 2026-04-17
AI Technical Summary
Current statins are not very effective in treating atherosclerosis and need to be combined with dietary and lifestyle changes; there is a lack of effective new methods.
Aescin was used in the preparation of drugs to activate AMPK, downregulate mTOR phosphorylation, induce autophagy in macrophage-derived foam cells and upregulate ABCA1 protein expression, and promote cholesterol efflux.
By activating AMPK, downregulating mTOR phosphorylation, and inducing autophagy, aescin can effectively reduce cholesterol accumulation in foam cells and improve atherosclerosis.
Smart Images
Figure CN121868322A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of natural compound application technology, specifically relating to new uses of aescin. Background Technology
[0002] Cardiovascular diseases (CVD) seriously endanger human health, causing approximately 17.9 million deaths annually, accounting for 31% of global deaths. Atherosclerosis (AS) is the leading cause of CVD morbidity and mortality. The pathological basis of CVD is lipid metabolism disorder, characterized by lesions in the affected arteries starting from the vascular intima, manifesting as lipid and carbohydrate accumulation, hemorrhage, and thrombosis. In early CVD, low-density lipoprotein (LDL) is oxidized to form oxidized low-density lipoprotein (ox-LDL), which is subsequently taken up by macrophages. The continuous influx of cholesterol exceeds the metabolic capacity of phagocytes, leading to intracellular accumulation and the formation of lipid droplets, thus transforming macrophages into cholesterol-rich foam cells. The continuously accumulating foam cells form lipid streaks on the vessel wall, gradually developing into a fibrous cap necrosis core, ultimately forming atherosclerotic plaques.
[0003] Current treatments for atherosclerosis include medication and surgery. Medication primarily works by lowering blood lipids, acting as an antiplatelet agent, and preventing blood clotting. Over the past few decades, the treatment of atherosclerosis has made some progress due to the use of statins and changes in diet and lifestyle. Nevertheless, a significant number of people do not respond well to statins, therefore, new methods to combat atherosclerosis are still needed.
[0004] Aescin is a sodium saponin salt extracted from the dried, mature seeds of the Chinese medicinal herb *Salvia splendens*, and is a natural herbal medicine. *Salvia splendens* is one of the Chinese medicinal materials included in the 2005 *Chinese Pharmacopoeia*, and its main functions are to regulate qi, relieve chest tightness, harmonize the stomach, and alleviate pain; it can be used for chest and abdominal distension and stomach pain. The main chemical components of *Salvia splendens* include aescin, flavonoids, oleic acid and stearic acid glycerides, polysaccharides, and proteins. Aescin is a mixture of more than 30 saponins extracted from *Salvia splendens*, mainly existing in the forms of α-aescinoside and β-aescinoside. Modern pharmacological studies have shown that aescin can promote the scavenging of oxygen free radicals and inhibit angiogenesis. Commercially available sodium aescin tablets are mainly used to treat venous edema. Existing research on aescin mainly focuses on its biological activities in treating gastric diseases and improving lower extremity venous edema, but research on its anti-atherosclerotic effects is rarely reported. Summary of the Invention
[0005] To address the shortcomings of the existing technology, this invention provides a new use for aescin, aiming to solve the technical problem that current treatments for atherosclerosis rely on statins, which require dietary and lifestyle changes. However, statins are not very effective in lowering lipids, and new methods to combat atherosclerosis are still needed.
[0006] The novel application of aescin provided by this invention is specifically illustrated by the following technical solution:
[0007] Application of aescin in the preparation of drugs for cardiovascular diseases.
[0008] In some embodiments, the aescin is used in the preparation of atherosclerotic drugs.
[0009] Furthermore, the aescin is used to prepare a drug that induces autophagy in macrophage-derived foam cells and promotes cholesterol efflux from foam cells.
[0010] Furthermore, the aescin is used to prepare a drug that induces macrophage-derived foam cell autophagy by activating AMPK and downregulating mTOR phosphorylation.
[0011] Furthermore, the aescin is used to prepare a drug that upregulates the expression of ABCA1 protein in macrophage-derived foam cells by activating AMPK and downregulating mTOR phosphorylation.
[0012] In some embodiments, the dosage form of the drug is an oral liquid, solid beverage, granules, capsules, tablets, effervescent tablets, syrup, pills, paste, ointment, emulsion, powder, microemulsion, powder for injection, water for injection, injection, nebulizer, gel, or nanoformulation.
[0013] In some embodiments, the drug is formulated as a sustained-release formulation, a controlled-release formulation, or a targeted formulation.
[0014] The present invention has the following beneficial effects: The present invention applies aescin to the preparation of drugs for atherosclerosis in cardiovascular diseases. Due to the reduction of AMPK phosphorylation, p-mTOR expression is increased, and the reduction of autophagy leads to the accumulation of cholesterol in macrophage-derived foam cells, which ultimately promotes the occurrence and development of atherosclerosis. In this application, aescin activates AMPK, downregulates mTOR phosphorylation to induce autophagy in macrophage-derived foam cells, and upregulates ABCA1 protein expression, thereby promoting cholesterol efflux from macrophage-derived foam cells and thus improving atherosclerosis. Attached Figure Description
[0015] Figure 1This is a graph showing the effect of aescin on the increase in total cholesterol and total triglyceride levels in RAW264.7 macrophages induced by ox-LDL stimulation, as described in Example 1 of this invention.
[0016] Figure 2 This is a graph showing the effect of aescin on the expression of mTOR protein in RAW264.7 macrophage-derived foam cells in Example 2 of the present invention;
[0017] Figure 3 This is a graph showing the effect of aescin on the LC3 II / LC3I ratio of RAW264.7 macrophage-derived foam cells in Example 3 of the present invention;
[0018] Figure 4 This is a graph showing the effect of aescin on the expression of ABCA1 protein in RAW264.7 macrophage-derived foam cells in Example 4 of the present invention;
[0019] Figure 5 This is a schematic diagram of the planar structure showing the effect of aescin on the expression of ABCA1 protein in RAW264.7 macrophage-derived foam cells in Example 4 of the present invention.
[0020] Figure 6 This is a schematic diagram illustrating the anti-atherosclerosis mechanism of aescin in this invention through the AMPK / mTOR / ABCA1 axis. Detailed Implementation
[0021] To make the objectives, technical solutions, and advantages of the present invention clearer, the present invention will be further described in detail below with reference to specific embodiments and accompanying drawings.
[0022] Example 1
[0023] Cholesterol accumulation within inflammatory cells is a significant factor promoting the occurrence and development of atherosclerosis. As early as 1913, Nikolai Anitschkow demonstrated that cholesterol intake induces atherosclerosis in rabbits and concluded that "without cholesterol, there is no atherosclerosis." Therefore, targeting cholesterol transport within macrophages is an effective approach to treating atherosclerosis.
[0024] RAW264.7 macrophages were incubated with ox-LDL (100 μg / mL) for 24 hours, followed by treatment with aescin (10, 20, and 40 μg / mL) for 12 hours. Figure 1 A) The levels of intracellular total cholesterol and total triglycerides were detected using a TC and TG detection kit. Figure 1 B). For example Figure 1As shown, with increasing aescin concentration, the intracellular total cholesterol and total triglyceride levels significantly decreased. This demonstrates that aescin (10, 20, and 40 μg / mL) can inhibit ox-LDL-induced increases in total cholesterol and total triglyceride levels in RAW264.7 macrophages.
[0025] The above experiments revealed that aescin can reduce the cholesterol content in macrophage-derived foam cells, demonstrating that aescin affects cholesterol transport in macrophage-derived foam cells, suggesting that aescin has a potential anti-atherosclerotic effect.
[0026] Example 2
[0027] AMP-activated protein kinase (AMPK), a crucial regulator of bioenergy metabolism, participates in various physiological processes. Studies have shown that upregulation of AMPK phosphorylation levels can slow the formation of atherosclerosis. Many drugs used to treat diabetes and atherosclerosis, such as metformin, thiazolidinediones, and statins, exert their angiprotective effects by activating AMPK.
[0028] Transcriptomic analysis was performed on the cells in Example 1. The intersection of differentially expressed genes between the RAW264.7 macrophage-derived foam cell model group and the aescin intervention group was extracted, and a heatmap of the expression abundance of the intersection genes in the two groups was plotted. Figure 2 A represents the heatmap analysis of differentially expressed genes between the AW264.7 macrophage-derived foam cell model group and the aescin intervention group (40 μg / mL). It shows that there are differences in the expression levels of genes related to phosphorylated AMPK protein between the macrophage-derived foam cell model group and the aescin intervention group (40 μg / mL). Figure 2 B represents the effect of aescin on phosphorylated AMPK and total AMPK protein expression in RAW264.7 macrophage-derived foam cells using Western blot analysis. The results show that Western blot experiments on macrophage-derived foam cells confirmed at the protein level that aescin (20 and 40 μg / mL) can activate AMPK signaling in RAW264.7 macrophage-derived foam cells.
[0029] Transcriptomic analysis, through intersection analysis of differentially expressed genes in the ox-LDL-induced RAW264.7 macrophage-derived foam cell model group and the aescin intervention group, revealed that aescin significantly upregulated AMPK gene expression in macrophage-derived foam cells. Further Western blot experiments validated this result at the cellular level. This suggests that aescin possesses a potent AMPK signaling activation effect, further demonstrating its potential anti-atherosclerotic activity.
[0030] Example 3
[0031] Upregulation of mTOR (Mammalian target of rapamycin) and p-mTOR proteins was observed in macrophage-derived foam cells, and blocking mTOR expression with specific siRNA inhibited foam cell formation. Qian Cao et al. also found that activation of the AMPK / mTOR pathway can repair macrophage autophagy, reduce the formation of foam cells derived from macrophages and vascular smooth muscle cells, thereby improving atherosclerosis.
[0032] Figure 3 A is a heatmap analysis of the differentially expressed genes between the RAW264.7 macrophage-derived foam cell model group and the aescin intervention group (40 μg / mL). It can be seen from the heatmap analysis that when the differentially expressed genes between the RAW264.7 macrophage-derived foam cell model group and the aescin intervention group were analyzed, the levels of genes related to phosphorylated mTOR protein expression were found to be different. Figure 3 B represents the effect of aescin on phosphorylated mTOR and mTOR protein expression in RAW264.7 macrophage-derived foam cells using Western blot analysis. The results show that Western blot experiments on macrophage-derived foam cells confirmed at the protein level that aescin (20, 40 μg / mL) can downregulate the expression of phosphorylated mTOR in macrophage-derived foam cells.
[0033] Similarly, using transcriptomics analysis, by performing intersection analysis on differentially expressed genes in the ox-LDL-induced RAW264.7 macrophage-derived foam cell model group and the aescin intervention group, it was found that aescin downregulated the expression of the mTOR gene in macrophage-derived foam cells. At the same time, Western blot results showed that aescin could inhibit the phosphorylation of mTOR in macrophage-derived foam cells.
[0034] Example 4
[0035] Studies have shown that autophagy dysfunction is a significant cause of atherosclerosis, and AMPK has been found to repair autophagy and inhibit foam cell formation, thereby improving atherosclerosis. Meanwhile, mTOR, a key molecule affecting autophagy, shows that downregulation of mTOR phosphorylation indicates autophagy activation. Furthermore, autophagy participates in macrophage reverse cholesterol transport, regulating the transfer of lipid droplets to macrophage-derived foam cell lysosomes. Lysosomal degradation of lipids leads to the efflux of free cholesterol, thus contributing to the regression of atherosclerotic plaques.
[0036] like Figure 4 As shown, Western blot experiments confirmed that aescin (10, 20 and 40 μg / mL) increased the ratio of autophagy-related proteins LC3 II / LC3 I on RAW264.7 macrophage-derived foam cells in a concentration-dependent manner, demonstrating that aescin has an activating effect on autophagy in RAW264.7 macrophage-derived foam cells.
[0037] Therefore, autophagy is a potential target for promoting cellular cholesterol efflux and preventing atherosclerotic cardiovascular disease. Upregulation of the LC3 II / LC3 I ratio can serve as a marker of autophagy activation. Western blot results showed that aescin can increase the LC3 II / LC3 I ratio of autophagy-related proteins on macrophage-derived foam cells, thus demonstrating that aescin induces autophagy in macrophage-derived foam cells. These results suggest that aescin can activate AMPK, inhibit downstream mTOR phosphorylation, and induce autophagy.
[0038] like Figure 5 As shown, RAW264.7 macrophage-derived foam cells were treated in the same way (with different concentrations of aescin), and Western blot results showed that aescin at concentrations of 20 and 40 μg / mL upregulated ABCA1 protein expression.
[0039] Recent studies have shown that a specific form of autophagy—lipophagy—can transport cholesterol esters (CE) from macrophages to lysosomes. After being degraded by lysosomal lipases, CE is transported to high-density lipoprotein (HDL) via the ATP-binding cassette transporter A1 (ABCA1), causing cholesterol efflux from macrophages. Western blot results showed that aescin increases ABCA1 protein expression in macrophage-derived foam cells.
[0040] In summary, the following conclusions can be drawn (e.g. Figure 6As shown in the figure: Decreased AMPK phosphorylation induces increased p-mTOR expression, and reduced autophagy leads to cholesterol accumulation in foam cells, ultimately promoting the development of atherosclerosis. Aescin induces autophagy in macrophage-derived foam cells by activating AMPK and downregulating mTOR phosphorylation, while upregulating ABCA1 protein expression, thereby promoting cholesterol efflux from macrophage-derived foam cells and improving atherosclerosis.
[0041] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. The present invention is not limited to the examples described above. Any changes, modifications, additions, or substitutions made by those skilled in the art within the scope of the present invention should also fall within the protection scope of the present invention.
Claims
1. Application of aescin in the preparation of cardiovascular disease drugs.
2. The application according to claim 1, characterized in that, The aescin was used in the preparation of drugs for atherosclerosis.
3. The application according to claim 2, characterized in that, The aescin was used to prepare a drug that induces autophagy in macrophage-derived foam cells and promotes cholesterol efflux from foam cells.
4. The application according to claim 2, characterized in that, The aescin was used to prepare a drug that induces macrophage-derived foam cell autophagy by activating AMPK and downregulating mTOR phosphorylation.
5. The application according to claim 2, characterized in that, The aescin was used to prepare a drug that upregulates ABCA1 protein expression in macrophage-derived foam cells by activating AMPK, downregulating mTOR phosphorylation.
6. The application according to claim 1, characterized in that, The dosage forms of the drug are oral liquids, solid beverages, granules, capsules, tablets, effervescent tablets, syrups, pills, pastes, ointments, emulsions, powders, microemulsions, powder injections, water injections, injections, nebulizers, gels, or nano-preparations.
7. The application according to claim 1, characterized in that, The drug is formulated as a sustained-release formulation, a controlled-release formulation, or a targeted formulation.