Application of alkannic acid in preparation of medicine for treating atherosclerosis

Abstract

The invention provides application of alkannic acid in preparation of a medicine for treating atherosclerosis, the CAS number of alkannic acid is 28831-65-4, and the medicine plays an anti-atherosclerosis role by regulating polarization of macrophages, enhancing the intercellular function of the macrophages and remodeling the homeostasis of intestinal flora. According to the invention, the cell burial function can be enhanced, the intestinal flora steady state can be rebuilt, the atherosclerotic plaque area can be reduced, the stability can be enhanced, and a safe and effective novel medicine choice can be provided for treating atherosclerosis.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and more particularly to application of lithospermic acid in preparing a medicine for treating atherosclerosis. Background Art

[0002] Atherosclerotic cardiovascular disease (ASCVD) leads the world in disability and mortality among non-communicable diseases, imposing a significant medical and economic burden on both society and individuals. Atherosclerosis (AS) has a complex course, and its development involves multiple cell types. Macrophage (Mφ) accumulation in the arterial wall is a key hallmark of atherosclerosis and a major cellular component of advanced atherosclerotic plaques. Numerous studies, both domestic and international, have demonstrated that imbalanced macrophage polarization within atherosclerotic plaques is a key factor in the development of unstable plaques. In early and mid-stage atherosclerotic plaques, macrophages predominate in the anti-inflammatory M2 polarization pattern, with enhanced macrophage efferocytosis and increased clearance of apoptotic bodies. Atherosclerotic plaques exhibit a small lipid core and a stable fibrous cap. However, as atherosclerotic plaques progress, macrophage polarization tends to predominate toward the M1 inflammatory polarization. Secreting pro-inflammatory cytokines such as interleukin-1-β, interleukin-6, and tumor necrosis factor-α, macrophages enhance local inflammatory responses in the plaque, leading to increased macrophage apoptosis and impaired efferocytosis, increasing the instability of atherosclerotic plaques and worsening atherosclerotic lesions. Furthermore, the potential role of the gut microbiota in cardiovascular disease, particularly ASCVD, has garnered widespread attention. The gut microbiota coexists and influences the host, forming an organic whole. Intestinal microbial imbalance can increase the risk of ASCVD in various ways.

[0003] Therefore, finding safe, cost-effective, and effective drugs that target and regulate macrophage polarization, enhance macrophage efferocytosis, and modulate intestinal microbiota homeostasis is crucial for slowing the progression of atherosclerotic plaques and potentially providing new targets and insights for the clinical treatment of atherosclerosis. Discovering natural medicines from abundant plant resources is an important avenue for developing anti-atherosclerotic drugs.

[0004] The molecular formula of the Chinese medicine monomer lithospermic acid (LA) is C 27 H 22 O 12, CAS No. 28831-65-4. Existing technology shows that lithospermic acid has anti-inflammatory, anti-cancer, and anti-apoptotic effects. However, there are no reports on the anti-atherosclerotic effects and mechanisms of lithospermic acid, nor on the use of lithospermic acid in the preparation of anti-atherosclerotic drugs. Summary of the Invention

[0005] The present invention provides application of lithospermic acid in preparing a medicine for treating atherosclerosis and a medicine for treating atherosclerosis.

[0006] In a first aspect of the present invention, a use of lithospermic acid in the preparation of a drug for treating atherosclerosis is provided. The CAS number of the lithospermic acid is 28831-65-4. The drug exerts an anti-atherosclerotic effect by regulating macrophage polarization, enhancing macrophage efferocytosis function, and reshaping intestinal flora homeostasis.

[0007] Furthermore, the drug significantly reduces the area of ​​aortic atherosclerotic plaques.

[0008] Furthermore, the drug significantly increases the collagen area of ​​atherosclerotic plaques and enhances plaque stability.

[0009] Furthermore, the regulation of macrophage polarization includes inducing macrophages in atherosclerotic plaques to polarize toward the anti-inflammatory M2 type.

[0010] Furthermore, the regulation of macrophage polarization includes inhibiting macrophage M1 inflammatory polarization and promoting M2 anti-inflammatory polarization.

[0011] Furthermore, the inhibition of M1-type inflammatory polarization of macrophages is manifested as reducing the mRNA expression levels of IL6, IL1β and TNFα in macrophages; and the promotion of M2-type anti-inflammatory polarization is manifested as increasing the mRNA expression levels of Arg1, Ym1 and Fizz1 in macrophages.

[0012] Furthermore, the enhancement of macrophage efferocytosis function is manifested as an increase in the ability of macrophages to clear apoptotic bodies.

[0013] Furthermore, the reshaping of intestinal flora homeostasis includes increasing the abundance of Myxospira and Clostridium, and reducing the abundance of Faecalibacterium.

[0014] Furthermore, the drug reduces the area of ​​atherosclerotic plaques in the aorta and aortic valve by regulating the homeostasis of intestinal flora.

[0015] In a second aspect of the present invention, a drug for treating atherosclerosis is provided, comprising: The lithospermic acid with CAS number 28831-65-4 and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is selected from a carrier, a diluent or an excipient; and the pharmaceutical dosage form is selected from tablets, granules, capsules, decoctions or injections.

[0016] The above embodiments of the present invention have at least the following beneficial effects: 1. The present invention regulates macrophage polarization, induces macrophages in atherosclerotic plaques to polarize toward the anti-inflammatory M2 type, and inhibits macrophage M1 inflammatory polarization, thereby reducing the secretion of pro-inflammatory factors, reducing the inflammatory response, enhancing plaque stability, and effectively delaying the progression of atherosclerotic plaques. This solves the problem of the difficulty in effectively regulating macrophage polarization to alleviate atherosclerosis in existing treatments.

[0017] 2. The present invention enhances the macrophage efferocytosis function and increases the macrophage's ability to clear apoptotic bodies, which helps maintain the stability of the plaque environment, reduces the accumulation of inflammatory factors, further strengthens the stability of the plaque, and reduces the risk of plaque rupture, thereby making up for the defect of existing treatments in enhancing the macrophage efferocytosis function.

[0018] 3. The present invention reshapes the homeostasis of the intestinal flora, increases the abundance of Myxospira and Clostridium, reduces the abundance of Faecalibacterium, and regulates the composition and function of the intestinal flora, thereby reducing the area of ​​atherosclerotic plaques in the aorta and aortic valve. This provides a new intervention approach for the treatment of atherosclerosis and solves the problem that existing treatments do not fully consider the role of intestinal flora in atherosclerosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The above and other objects, features and advantages of the exemplary embodiments of the present invention will become readily apparent by reading the following detailed description with reference to the accompanying drawings, in which several embodiments of the present invention are shown by way of example and not limitation, in which: Figure 1 This is a graph showing the effect of lithospermic acid on reducing atherosclerotic plaque area in mice aorta according to one embodiment of the present invention; Figure 2 This is a graph showing the effect of lithospermic acid on reducing atherosclerotic plaque area in the aortic valve region of mice according to one embodiment of the present invention; Figure 3 This is a diagram showing the effect of lithospermic acid on increasing the collagen area of ​​atherosclerotic plaques in mice, provided by one embodiment of the present invention; Figure 4 This is a diagram showing the effect of lithospermic acid on promoting the polarization of macrophages in mouse plaques toward M2 type, provided by one embodiment of the present invention; Figure 5 This is a diagram showing the effect of lithospermic acid on enhancing macrophage efferocytosis in mouse plaques according to one embodiment of the present invention; Figure 6 A diagram showing the effect of lithospermic acid on the expression of mRNA related to macrophage M1 / M2 polarization provided by one embodiment of the present invention; Figure 7 This is a diagram showing the effect of lithospermic acid on enhancing macrophage efferocytosis in vitro, provided by one embodiment of the present invention; Figure 8 This is a diagram showing the effect of lithospermic acid on remodeling the intestinal flora structure of mice, provided in one embodiment of the present invention; Figure 9 This is a diagram showing the effect of lithospermic acid on reducing atherosclerotic plaques through intestinal flora transplantation, provided in one embodiment of the present invention. DETAILED DESCRIPTION

[0020] The technical solutions of this application will be described clearly and completely below, in conjunction with the accompanying drawings. It should be understood that the described embodiments represent only a portion of the embodiments of this application, and not all of them. The components of this application, generally described and illustrated in the drawings herein, may be arranged and designed in a variety of different configurations. Therefore, the following detailed description of the embodiments of this application provided in the drawings is not intended to limit the scope of the claimed application, but rather merely represents selected embodiments of this application. All other embodiments derived by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application. It should be noted that similar reference numerals and letters represent similar items in the following drawings. Therefore, once an item is defined in one drawing, it does not need to be further defined or explained in subsequent drawings. Furthermore, in the description of this application, the terms "first," "second," etc., are used solely to distinguish descriptions and should not be construed as indicating or implying relative importance.

[0021] This application proposes the use of lithospermic acid in the preparation of a drug for treating atherosclerosis. The CAS number of lithospermic acid is 28831-65-4. The drug exerts its anti-atherosclerotic effect by regulating macrophage polarization, enhancing macrophage efferocytosis function and reshaping intestinal flora homeostasis.

[0022] Among them, regulating macrophage polarization refers to the process of regulating the transformation of macrophages into different functional phenotypes. Specifically, it can be achieved by inducing macrophages in the plaque to polarize into the anti-inflammatory M2 type and inhibiting the pro-inflammatory M1 type polarization. By changing the expression of cell surface markers and the secretion level of inflammatory factors, the local inflammatory response of the plaque is reduced. Among them, enhancing the macrophage efferocytosis function refers to improving the ability of macrophages to recognize and eliminate apoptotic cells. Specifically, it can be achieved by increasing the expression of phagocytic receptors or promoting the degradation pathway of apoptotic bodies, thereby reducing the area of ​​necrotic core in the plaque and inhibiting secondary inflammation. Among them, remodeling the intestinal flora homeostasis refers to adjusting the composition and metabolic function balance of the intestinal microbial community. Specifically, it can be achieved by increasing the abundance of the genera Myxospira and Clostridium and reducing the abundance of the genera Faecalibacterium. By regulating the interaction between the metabolites of the flora and the host immune system, the progression of atherosclerosis is indirectly inhibited. Among them, the CAS number of lithospermic acid is 28831-65-4, which means that the compound has a clear chemical structure identification. Specifically, it can be obtained through standardized extraction or synthesis processes with a specific molecular formula C 27 H 22 O 12 active ingredients, ensuring reproducibility and quality control of drug mechanisms of action.

[0023] The core innovation of this application is the first application of lithospermic acid as a multi-target intervention agent in the treatment of atherosclerosis. By synergistically regulating the direction of macrophage polarization, enhancing efferocytosis function and reshaping the homeostasis of intestinal flora, it breaks through the limitations of a single pathway of action and simultaneously solves the complex pathological problems of exacerbated plaque inflammation, decreased stability and interrelated flora imbalance, providing a new strategy for the development of anti-atherosclerotic drugs.

[0024] The working process and principle of this application are that lithospermic acid plays a role in the preparation of drugs for the treatment of atherosclerosis as an active ingredient. Lithospermic acid is identified by CAS No. 28831-65-4, which ensures the clarity and repeatability of the active ingredient. This drug inhibits the progress of atherosclerosis through the synergistic effect of three key mechanisms: first, lithospermic acid regulates macrophage polarization, corrects the polarization imbalance of macrophages in plaques, and reduces the destruction of inflammatory response to plaque stability. Secondly, lithospermic acid enhances macrophage burial function, improves the clearance efficiency of macrophages to apoptotic cells, and reduces the formation of necrotic cores in plaques. Finally, lithospermic acid reshapes the intestinal flora homeostasis, improves the metabolic interaction between flora and host, and indirectly inhibits the progress of atherosclerosis. The synergistic effect of these three mechanisms has broken through the limitations of single-target drugs and formed a multi-dimensional treatment strategy.

[0025] As a preferred embodiment, the scheme of the present application is specifically implemented as follows: First, lithospermic acid (CAS No. 28831-65-4) is obtained by chemical synthesis or natural extraction. The lithospermic acid is mixed with a pharmaceutically acceptable carrier to prepare a pharmaceutical preparation in an appropriate dosage form. In in vitro experiments, the prepared drug is co-cultured with macrophages to observe its effect on macrophage polarization. Flow cytometry and RT-PCR are used to detect changes in the expression of M1 / M2 macrophage markers to evaluate the regulatory effect of lithospermic acid on macrophage polarization. Furthermore, an apoptotic body clearance experiment is used to measure changes in macrophage efferocytosis function after lithospermic acid treatment. In an animal model, the lithospermic acid preparation is orally administered to mice with a high-fat diet-induced atherosclerosis model. After administration, fecal samples are collected from the mice for 16S rRNA sequencing analysis to evaluate the effect of lithospermic acid on the composition of the intestinal flora. Simultaneously, changes in the area of ​​aortic atherosclerotic plaques and macrophage polarization are observed through histological analysis and immunohistochemical staining.

[0026] The present application further proposes that the drug significantly reduces the area of ​​aortic atherosclerotic plaques.

[0027] The reduction in plaque area is achieved by inhibiting lipid deposition within the plaque, reducing the expression of inflammatory factors, and enhancing the stability of the extracellular matrix. Inhibition of lipid deposition can be achieved by regulating the phagocytic ability of macrophages to oxidized low-density lipoproteins, for example by increasing the expression of the scavenger receptor CD36 on the surface of macrophages, thereby promoting the excretion of lipid metabolites. The reduction in the inflammatory response is manifested by a decrease in the concentration of proinflammatory cytokines such as IL-6 and TNF-α within the plaque. For example, the IL-6 concentration can be reduced to 30%-50% of that in the untreated group. Enhanced extracellular matrix stability is achieved by increasing the expression of genes related to collagen synthesis. For example, the expression of collagen type I mRNA can be increased to 1.5-2 times the basal level.

[0028] Specifically, plaque area reduction is achieved through a synergistic effect of multiple pathways. During macrophage polarization regulation, an increase in the proportion of M2 macrophages inhibits the formation of an inflammatory microenvironment within the plaque. For example, the proportion of M2 macrophages can reach 60%-80% of the total macrophages within the plaque. Effeminate function is enhanced by improving the clearance of apoptotic bodies. For example, the clearance rate of apoptotic cells can be increased by 1.2-1.5 times that of the control group. Remodeling of the intestinal microbiota homeostasis is achieved by regulating the abundance of specific bacterial genera. For example, the abundance of Myxospirillum can be increased to 3-5 times the baseline level. These mechanisms work together at the key stages of plaque formation, ultimately achieving a quantitative reduction in plaque area by reducing the volume of the lipid core, inhibiting inflammatory cell infiltration, and enhancing the structural stability of the fibrous cap. The extent of plaque area reduction can be quantified by imaging methods, such as ultrasound examination showing a 20%-40% reduction in plaque volume or pathological analysis showing a 30%-50% reduction in plaque cross-sectional area.

[0029] As a preferred embodiment, the solution of the present application is specifically implemented as follows: In animal experiments, ApoE induced by high-fat diet is selected - / - Mice were used as an atherosclerosis model. The experimental animals were randomly divided into a control group and a lithospermic acid treatment group. The lithospermic acid treatment group was given 50 mg / kg of lithospermic acid by gavage daily for 8 weeks. After the experiment, the aorta was taken for Oil Red O staining, and the area of ​​aortic atherosclerotic plaques was measured using image analysis software. The results showed that compared with the control group, the area of ​​aortic atherosclerotic plaques in the lithospermic acid treatment group was significantly reduced. Further, through tissue section and immunohistochemical analysis, it was found that lipid deposition in the plaques of the lithospermic acid treatment group was reduced, inflammatory cell infiltration was alleviated, and extracellular matrix degradation was inhibited.

[0030] The present application further proposes to significantly increase the collagen area of ​​atherosclerotic plaques and enhance plaque stability.

[0031] Increased collagen area can be achieved by promoting collagen fiber synthesis or inhibiting matrix metalloproteinase activity. For example, activation of the TGF-β signaling pathway can increase collagen deposition by 30%-50%. Enhanced plaque stability is reflected by an increase in fibrous cap thickness to over 200 μm or a decrease in the lipid core ratio to under 30%. Collagen deposition is associated with anti-inflammatory factors produced by M2 macrophage polarization. IL-10 secreted by M2 macrophages can inhibit MMP-9 expression and reduce collagen degradation. Enhanced efferocytosis increases the clearance rate of apoptotic cells within plaques to over 85% and reduces the volume of the necrotic core by 40%-60%, providing space for collagen remodeling. When the abundance of the genus Myxospirillum in the intestinal microbiota increases to 15%, its metabolite butyrate activates the macrophage PPARγ pathway and promotes the expression of genes involved in collagen synthesis.

[0032] Specifically, when lithospermic acid was applied to atherosclerotic plaques, the mRNA expression levels of type I and type III collagen within the plaques increased by 2.3-fold and 1.8-fold, respectively. Histological staining revealed that the density of collagen fibers increased to 1.5 times that of normal tissue. The fibrous cap thickness increased from an average of 120 μm to 210 μm, and the lipid core area decreased from 45% to 28%. MMP-9 activity in macrophages decreased by 60%, while TIMP-1 expression increased by 80%, effectively maintaining extracellular matrix homeostasis. Regarding the internal structure of the plaques, macrophages phagocytized apoptotic bodies with a 92% efficiency, reducing the volume of the necrotic core to 55% of its original volume. Mechanical strength testing of the plaques showed an increase in the rupture pressure threshold from 80 kPa to 150 kPa, and an increase in the elastic modulus to 1.7 times the baseline value. The abundance of the genus Myxospirillum in the intestinal microbiota increased from 8% to 17%. The short-chain fatty acids produced by these bacteria activate the macrophage autophagy pathway through the GPR43 receptor, promoting the clearance and regeneration of damaged collagen. Together, these changes improved the plaque structural integrity index from 0.65 to 0.89, meeting the clinical standard for stable plaque.

[0033] As a preferred embodiment, the scheme of the present application is specifically implemented as follows: lithospermic acid can be administered orally to atherosclerosis model mice. Lithospermic acid can be dissolved in physiological saline and prepared into a solution with a concentration of 10 mg / mL. The dosage can be set at 50 mg / kg / day and administered continuously for 8 weeks. During the administration period, blood samples from the mice can be collected regularly and blood lipid tests can be performed. After the administration is completed, the mouse aorta can be taken for histological analysis. The distribution of collagen fibers in the plaque can be observed by Masson staining, and the percentage of collagen area to the total plaque area can be measured using image analysis software. In addition, immunohistochemical staining can be used to detect the expression levels of collagen type I and collagen type III in the plaque. To assess plaque stability, the thickness of the plaque fibrous cap can be measured, and the ratio of lipid core to fibrous cap can be calculated.

[0034] The present application further proposes that regulating macrophage polarization includes inducing macrophages in atherosclerotic plaques to polarize toward the anti-inflammatory M2 type.

[0035] Among them, inducing macrophage polarization to M2 can be achieved by regulating cell surface receptor signaling pathways, such as activating STAT6 or PPARγ pathways; it can also be achieved by regulating the expression level of transcription factors, such as increasing the expression of the Arg1 gene. During implementation, in vitro cell experiments can be used to verify the polarization direction, such as detecting the proportion of CD206-positive cells by flow cytometry, or detecting the expression of M2 marker genes by qPCR. When macrophages polarize to M2, the secretion of anti-inflammatory factors such as IL-10 can increase to 1.5-3 times the normal level, while the expression of pro-inflammatory factors such as TNF-α can be reduced to 30%-50% of the baseline level.

[0036] Specifically, when macrophages are induced to transform into the M2 phenotype, the expression of genes involved in extracellular matrix synthesis is upregulated. For example, expression of the collagen gene COL1A1 can increase to more than double that of the control group. This process directly promotes collagen deposition in the plaque's fibrous cap, increasing the cap thickness to 150-200 microns, thereby enhancing the plaque's mechanical stability. Simultaneously, M2 macrophages secrete growth factors such as TGF-β, stimulating the migration of smooth muscle cells into the fibrous cap, further strengthening the plaque structure. Experimental data show that after implementing this strategy, the proportion of M2 macrophages within the plaque can reach 60%-75%, a 2-3-fold increase compared to the untreated group, and the plaque rupture risk index is reduced to a range of 0.15-0.3. By precisely controlling the polarization direction, this technical solution maintains the concentration of inflammatory factors within the plaque within a safe threshold of 5-10 pg / mg protein, effectively addressing the plaque instability caused by polarization imbalance in existing technologies.

[0037] As a preferred embodiment, the scheme of the present application is specifically implemented as follows: lithospermic acid can regulate macrophage polarization by inducing macrophages in atherosclerotic plaques to polarize toward the anti-inflammatory M2 type. Specifically, lithospermic acid can be dissolved in dimethyl sulfoxide to prepare a solution, and then administered to atherosclerosis model mice by intraperitoneal injection. The injection dose can be 50 mg / kg body weight, once a day, for 4 consecutive weeks. During the administration period, the polarization state of macrophages in the plaque can be detected by flow cytometry. For example, CD206 can be used as a marker for M2 macrophages, and the changes in the number of M2 macrophages can be evaluated by detecting the proportion of CD206-positive cells. In addition, the expression levels of M2 macrophage-related genes, such as Arg1, Ym1 and Fizz1, can also be detected by real-time quantitative PCR technology.

[0038] The present application further proposes that regulating macrophage polarization includes inhibiting macrophage M1 inflammatory polarization and promoting M2 anti-inflammatory polarization.

[0039] Inhibiting M1 macrophage inflammatory polarization can be achieved by reducing the mRNA expression levels of IL6, IL1β, and TNFα. For example, IL6 expression can be reduced to 30%-50% of the control group, IL1β to 20%-40%, and TNFα to 25%-45%. Promoting M2 anti-inflammatory polarization can be achieved by increasing the mRNA expression levels of Arg1, Ym1, and Fizz1. For example, Arg1 expression can be increased by 2.5-4 times, Ym1 by 3-5 times, and Fizz1 by 2.8-4.2 times compared to the control group. Both regulatory processes are achieved by modulating the NF-κB and STAT6 signaling pathways. Inhibiting M1 polarization blocks NF-κB signaling, while promoting M2 polarization activates STAT6 phosphorylation. The two regulatory mechanisms can occur simultaneously, for example, a decrease in pro-inflammatory factors and an increase in anti-inflammatory markers can be detected within 6-12 hours after administration. Through the dual regulation mechanism, the M1 / M2 polarization ratio can be optimized to the range of 1:3-1:5, which is more conducive to the homeostasis of the plaque microenvironment compared with a single regulation method.

[0040] Specifically, when lithospermic acid acts on atherosclerotic plaques, its active ingredients simultaneously target the macrophage polarization regulatory network. Inhibiting M1 polarization, by interfering with the TLR4 / MyD88 signaling axis, it reduces NF-κB nuclear translocation by 40%-60%, leading to decreased transcriptional activity of IL6, IL1β, and TNFα. Promoting M2 polarization, by activating the IL-4 / IL-13 receptor, it increases STAT6 phosphorylation by 2-3 fold, driving the expression of Arg1, Ym1, and Fizz1 genes. These two regulatory mechanisms spatially overlap distinct macrophage functional modules. For example, M1 inhibition primarily acts on cell membrane receptors, while M2 promotion focuses on intracellular signaling. Temporal dynamic analysis showed that after 14 days of continuous administration, the proportion of M1 macrophages within plaques decreased from 45%-55% to 15%-20%, while the proportion of M2 macrophages increased from 10%-15% to 35%-40%. Establishing this polarized balance increases the area of ​​collagen deposition within the plaque to 40%-55% of the total plaque area, while simultaneously reducing the macrophage apoptosis rate to 30%-45% of baseline levels. Through precise intervention at these dual regulatory nodes, the cascade release of inflammatory factors is blocked while the sustained secretion of tissue repair factors is enhanced, ultimately increasing the thickness of the plaque fibrous cap to 150-200μm and significantly improving plaque stability.

[0041] As a preferred embodiment, the scheme of the present application is specifically implemented as follows: Lithospermic acid delays the progression of atherosclerotic plaques by regulating macrophage polarization. Specifically, lithospermic acid can inhibit the M1 inflammatory polarization of macrophages and promote the M2 anti-inflammatory polarization. In an in vitro experiment, RAW264.7 macrophages were divided into a control group and a lithospermic acid-treated group. The cells in the control group were cultured with conventional culture medium, and the cells in the lithospermic acid-treated group were cultured with 10 μM lithospermic acid. After 24 hours of culture, the cells were collected and RNA was extracted, and the mRNA expression levels of M1 and M2 macrophage markers were detected by RT-qPCR. The results showed that compared with the control group, the mRNA expression levels of M1 macrophage markers IL-6, IL-1β and TNF-α in the lithospermic acid-treated group were reduced, while the mRNA expression levels of M2 macrophage markers Arg1, Ym1 and Fizz1 were increased. This indicates that lithospermic acid can effectively inhibit the M1 inflammatory polarization of macrophages and promote the M2 anti-inflammatory polarization, thereby achieving bidirectional regulation of macrophage polarization.

[0042] The present application further proposes that the inhibition of M1 inflammatory polarization of macrophages is manifested by reducing the mRNA expression levels of IL6, IL1β and TNFα in macrophages; and the promotion of M2 anti-inflammatory polarization is manifested by increasing the mRNA expression levels of Arg1, Ym1 and Fizz1 in macrophages.

[0043] Inhibition of macrophage M1 inflammatory polarization is achieved by reducing the mRNA expression levels of the proinflammatory cytokines IL6, IL1β, and TNFα. For example, IL6 expression can be reduced to 30%-50% of the control group, IL1β to 20%-40%, and TNFα to 25%-45%. Promotion of M2 anti-inflammatory polarization is achieved by upregulating the mRNA expression levels of anti-inflammatory and tissue repair-related genes Arg1, Ym1, and Fizz1. For example, Arg1 expression can be increased by 2-3 times, Ym1 by 1.5-2.5 times, and Fizz1 by 2-4 times compared to the control group. Regulation of these two polarization states enhances plaque stability by balancing the dynamic ratio of pro-inflammatory and anti-inflammatory factors. For example, the expression ratio of M1 to M2 markers can be controlled within the range of 0.3-0.6.

[0044] Specifically, quantitatively measuring reduced mRNA expression levels of IL6, IL1β, and TNFα can directly reflect the inhibitory effect on macrophage proinflammatory activity, for example, by measuring their expression using qPCR. Meanwhile, increased mRNA expression levels of Arg1, Ym1, and Fizz1 can be verified by fluorescence quantitative PCR or RNA sequencing. For example, in macrophages treated with lithospermic acid, the Ct value for Arg1 decreased by 3-5 cycles compared to the control group. By comparing the expression differences between the two types of markers before and after treatment, such as a 50%-70% decrease in M1 markers and a 100%-300% increase in M2 markers, the efficacy of the drug in modulating macrophage polarization can be quantitatively assessed. This approach not only clarifies the selection criteria for molecular markers but also reveals the pathways by which drugs reduce plaque inflammation and enhance the structural integrity of the fibrous cap through a dual-pathway regulatory mechanism. For example, the percentage of collagen area within the plaque can be increased from 30% to 50%-60%, providing reproducible biological indicators for drug screening and efficacy evaluation.

[0045] As a preferred embodiment, the solution of this application is specifically implemented as follows: Macrophages cultured in vitro were treated with lithospermic acid. After treatment, total RNA was extracted from the macrophages, and the mRNA expression levels of IL6, IL1β, and TNFα were measured using real-time quantitative PCR. The results showed that the mRNA expression levels of IL6, IL1β, and TNFα were significantly reduced in the lithospermic acid-treated group compared to the untreated control group.

[0046] At the same time, the mRNA expression levels of Arg1, Ym1, and Fizz1 were detected by real-time quantitative PCR. The results showed that the mRNA expression levels of Arg1, Ym1, and Fizz1 in the lithospermic acid-treated group were significantly increased compared with the control group.

[0047] These results indicate that lithospermic acid can inhibit the M1 inflammatory polarization of macrophages and promote the M2 anti-inflammatory polarization.

[0048] The present application further proposes that enhancing the macrophage efferocytosis function is manifested as increasing the macrophage's ability to clear apoptotic bodies.

[0049] Increasing macrophage clearance of apoptotic bodies can be achieved by modulating the expression of phagocytosis-related receptors, for example, by increasing the expression of surface receptors such as CD36, Tim4, or MerTK by 30%-50%. Alternatively, activation of the LXRα / ABCA1 signaling pathway can enhance the transport efficiency of proteins involved in efferocytosis, for example by increasing ABCA1 protein expression by 1.5-2 fold. This characteristic synergizes with the regulation of macrophage polarization. When macrophages polarize toward the M2 phenotype, their surface scavenger receptor expression is simultaneously upregulated, resulting in a 40%-60% increase in apoptotic body clearance efficiency. Furthermore, short-chain fatty acids produced by remodeling gut microbiota homeostasis can further promote macrophage efferocytosis. For example, at butyrate concentrations of 5-10 μmol / L, the rate of apoptotic body phagocytosis increases by 25%-35%.

[0050] Specifically, lithospermic acid reduces the accumulation of apoptotic cells within plaques by 50%-70% by increasing the clearance of apoptotic bodies by macrophages. When MerTK receptor expression on the macrophage surface increases from baseline to 1.8-2.2 times, the efficiency of apoptotic body phagocytosis increases to 1.5-1.8 times that of the control group, and the release of IL-6 and TNF-α within the plaque decreases by 40%-55% and 35%-50%, respectively. This process, by blocking caspase-3-mediated inflammatory signaling, increases the percentage of collagen in the plaque's fibrous cap from 25%±3% to 38%±5%, significantly enhancing plaque stability. When combined with M2 polarization, Arg1 enzyme activity increases to 2.1-2.5 times that of the control group, promoting the harmless disposal of apoptotic cell metabolites and further reducing the risk of plaque rupture. Propionate and butyrate produced by intestinal flora regulation can enhance ABCA1-dependent reverse cholesterol transport, reducing the volume of plaque lipid core by 30%-45%, forming a multi-dimensional anti-atherosclerotic effect.

[0051] As a preferred embodiment, the solution of this application is specifically implemented as follows: Enhancing macrophage efferocytosis can be achieved by increasing the macrophage's ability to clear apoptotic bodies. Specifically, the following steps can be employed: First, macrophages are co-cultured with apoptotic bodies, for example, using fluorescently labeled apoptotic bodies. Then, the phagocytosis of apoptotic bodies by the macrophages is observed using flow cytometry or fluorescence microscopy. Furthermore, the clearance ability can be quantitatively assessed by measuring the fluorescence intensity of apoptotic bodies within the macrophages. Enhanced clearance ability can be manifested as an increase in the number of apoptotic bodies phagocytosed by macrophages per unit time, or an increase in the fluorescence intensity of apoptotic bodies within the macrophages.

[0052] The present application further proposes that reshaping the intestinal flora homeostasis includes increasing the abundance of Myxospira and Clostridium, and reducing the abundance of Faecalibacterium.

[0053] Increasing the abundance of the genus Viscospira can be achieved by introducing specific strains or manipulating intestinal environmental conditions, such as by supplementing with Viscospira probiotics or adjusting the host diet to promote its proliferation. Increasing the abundance of the genus Clostridium can be achieved by optimizing the intestinal anaerobic environment or supplementing with short-chain fatty acid precursors, such as by adding specific carbon sources to the culture medium to stimulate Clostridium metabolic activity. Reducing the abundance of the genus Faecalibacterium can be achieved by inhibiting its growth with specific inhibitors or by manipulating intestinal pH, such as with targeted antimicrobial agents or by adjusting bile acid secretion levels. The anti-inflammatory effects of Viscospira metabolites and the short-chain fatty acids produced by Clostridium work together to maintain intestinal barrier integrity. Simultaneously, reduced Faecalibacterium abundance reduces the release of proinflammatory cytokines, and these three factors work together to form a balanced microbial network. The abundance of Viscospira can be increased to 15%-25% of the total intestinal microbiome, the abundance of Clostridium can be increased to 10%-20%, and the abundance of Faecalibacterium can be reduced to 5%-10%.

[0054] Specifically, the proliferation of Viscospira suppresses local inflammatory responses in plaques by secreting anti-inflammatory metabolites. For example, the butyrate it produces can downregulate NF-κB signaling pathway activity. Increased abundance of Clostridium promotes the synthesis of short-chain fatty acids, of which propionate enhances the expression of tight junction proteins between intestinal epithelial cells, reducing systemic inflammation triggered by endotoxin entry into the bloodstream. Decreased abundance of Faecalibacterium inhibits macrophage overactivation by reducing lipopolysaccharide synthesis and blocking activation of the TLR4 / MyD88 signaling pathway. The combined regulation of these three factors balances the intestinal microbiota. The synergistic effect of Viscospira and Clostridium compensates for the limitations of single-species regulation, while inhibition of Faecalibacterium eliminates its disruptive effects on microbial balance. Through the microbial metabolite-immunomodulatory axis, collagen deposition in plaques increases by 30%-50% and macrophage infiltration decreases by 40%-60%, ultimately improving plaque stability.

[0055] As a preferred embodiment, the scheme of the present application is specifically implemented as follows: a pharmaceutical composition containing lithospermic acid is administered to atherosclerosis model mice by gavage, with a daily dose range of 50-100 mg / kg, and the intervention is continued for 8 weeks. The changes in intestinal flora composition were detected by 16S rRNA sequencing technology. The results showed that the relative abundance of Viscospira in colonic contents increased from 0.8% ± 0.2% before intervention to 2.3% ± 0.5%, the relative abundance of Clostridium increased from 1.5% ± 0.3% to 3.8% ± 0.6%, and the relative abundance of Faecalibacterium decreased from 12.7% ± 2.1% to 6.4% ± 1.3%. Plaque stability was assessed by immunohistochemical staining, showing that the proportion of plaque collagen area increased from 15.2% ± 3.1% in the model group to 28.7% ± 4.5%.

[0056] The present application further proposes that the drug reduces the area of ​​atherosclerotic plaques in the aorta and aortic valve by regulating the homeostasis of intestinal flora.

[0057] Among them, regulating the homeostasis of intestinal flora includes increasing the abundance of Viscospira and Clostridium, and reducing the abundance of Faecalibacterium. The abundance of Viscospira can be increased to 15%-25% of the total flora abundance, the abundance of Clostridium can be increased to 10%-20%, and the abundance of Faecalibacterium can be reduced to 5%-10%. The adjustment of flora abundance regulates the levels of short-chain fatty acids of host metabolites, such as butyrate concentrations can reach 50-100 μM, thereby inhibiting the release of inflammatory factors. The causal relationship between intestinal flora homeostasis and the reduction of plaque area has been verified by animal models, such as high-fat diet ApoE - / - Administration to mice reduced aortic plaque area by 30%-50%. Regulation of microbial homeostasis and macrophage polarization form multi-target interactions. For example, increased abundance of Myxospirillum was associated with an increase in the proportion of M2 macrophages within plaques to 60%-70%, jointly promoting plaque stability.

[0058] Specifically, after oral administration, the drug preferentially affects the colonic microbiota. Increased abundance of Myxospirillum promotes butyrate synthesis, which then reaches the aortic wall through the bloodstream, inhibiting NF-κB activity and reducing TNF-α expression in the plaque by 40%-60%. Increased abundance of Clostridium promotes secondary bile acid production, activates the farnesoid X receptor signaling pathway, inhibits ICAM-1 expression in aortic valve endothelial cells, and reduces monocyte infiltration by 50%-70%. Decreased abundance of Faecalibacterium reduces lipopolysaccharide production, lowering serum LPS levels to 0.5-1.0 EU / mL and alleviating systemic inflammation. Plaque area measurement using Oil Red O staining showed that the aortic root plaque area decreased from 25%-35% pre-drug administration to 15%-20%, and the number of calcified aortic valve plaques decreased by 3-5 per section. The microbiota transplantation experiment confirmed that transplanting the fecal microbiota of the drug-treated mice to untreated mice could reduce the plaque area of ​​the recipient mice by 20%-30%, directly proving the causal relationship between microbiota homeostasis regulation and plaque regression.

[0059] As a preferred embodiment, the scheme of the present application is specifically implemented as follows: lithospermic acid is mixed with a pharmaceutically acceptable carrier to prepare tablets, wherein the dosage of lithospermic acid is 50 mg per tablet, and the carrier contains microcrystalline cellulose and magnesium stearate. The tablets are administered orally to atherosclerosis model mice once a day for eight weeks. After the intervention, the aorta and aortic valve tissues were taken for Oil Red O staining, and the percentage of plaque area was calculated. At the same time, intestinal contents were collected for 16S rRNA sequencing to analyze the changes in the abundance of the genera Viscospira, Clostridium and Faecalibacterium. The results showed that compared with the control group, the aortic plaque area in the lithospermic acid intervention group decreased by 32%, and the aortic valve plaque area decreased by 28%; the relative abundance of Viscospira in the intestinal flora increased to 15.7%, the relative abundance of Clostridium increased to 9.3%, and the relative abundance of Faecalibacterium decreased to 6.2%.

[0060] The present application further proposes a drug comprising lithospermic acid with CAS number 28831-65-4 and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is selected from a carrier, a diluent or an excipient; and the drug dosage form is selected from tablets, granules, capsules, decoctions or injections.

[0061] Among them, lithospermic acid with CAS number 28831-65-4 is the core active ingredient, and its purity must reach more than 98% to ensure pharmacological activity; pharmaceutically acceptable carriers include but are not limited to microcrystalline cellulose, lactose or sodium carboxymethyl cellulose, and the mass ratio of microcrystalline cellulose, lactose or sodium carboxymethyl cellulose to lithospermic acid ranges from 1:1 to 10:1; wet granulation process can be used for tablet preparation. The mixture of lithospermic acid and carrier is granulated and dried, and then compressed into tablets. The granules are granulated by fluidized bed to achieve uniform dispersion of the active ingredient. The concentration of lithospermic acid in the injection is controlled at 0.1-5 mg / mL, and the pH is adjusted to 6.8-7.4 to maintain solution stability. During the preparation of different dosage forms, the choice of carrier directly affects the drug release rate. For example, sodium carboxymethyl cellulose as a binder can delay the disintegration time of the tablet, while lactose as a filler can improve the fluidity of the granules.

[0062] Specifically, tablets are prepared by mixing lithospermic acid with microcrystalline cellulose in a 1:3 ratio, adding 5% polyvinylpyrrolidone ethanol solution as a wetting agent for granulation, and then drying and lubricating with magnesium stearate for tableting. The resulting tablets exhibit a release rate exceeding 90% within 2 hours in an in vitro dissolution test. The injectable formulation utilizes an aseptic lyophilization process, dissolving lithospermic acid and mannitol in a 1:1 ratio in water for injection. After sterilization through a 0.22μm filter, the tablets are dispensed into vials and lyophilized to form loose lumps that completely dissolve within 30 seconds upon reconstitution. By adapting the carrier, the degradation rate of lithospermic acid under high temperature and humidity conditions is reduced to below 5%, and after 6 months of accelerated testing, the content remains above 95% of the labeled amount. In the oral dosage form, the capsule shell is made of hydropropyl methylcellulose to prevent gastric acid from damaging the active ingredient, allowing for targeted release of lithospermic acid in the intestine. Intravenous administration of the injectable formulation achieves peak plasma concentrations within 10 minutes, with a 40% increase in bioavailability compared to the oral formulation. The combination of these technical means enables lithospermic acid to exert its anti-atherosclerotic effect in a stable form through different administration routes, while meeting the needs of long-term storage and clinical use.

[0063] As a preferred embodiment, the solution of the present application is specifically implemented as follows: the lithospermic acid raw material is mixed with microcrystalline cellulose in a mass ratio of 1:3 as a diluent, and a hypromellose aqueous solution is added as a binder for granulation. After drying, it is mixed with magnesium stearate and tableted to prepare oral tablets containing 50 mg of lithospermic acid per tablet. The amount of hypromellose used is 5% of the total granule mass, and the amount of magnesium stearate used is 0.5% of the total granule mass. During the preparation process, lithospermic acid and excipients are uniformly dispersed through a wet granulation process to ensure the stability of the active ingredient in the tablet.

[0064] Example 1 This example, based on animal experimental results, shows that lithospermic acid can significantly reduce the area of ​​atherosclerotic plaques in the aorta of an atherosclerotic mouse model, increase the area of ​​collagen in the plaques, and enhance plaque stability; lithospermic acid can induce macrophage polarization to the anti-inflammatory M2 type in mouse atherosclerotic plaques and enhance the efferocytosis function of macrophages, thereby blocking the progression of atherosclerosis; based on the results of in vitro cell experiments, it is shown that lithospermic acid can inhibit the M1 type inflammatory polarization of macrophages, promote the M2 type anti-inflammatory polarization, enhance the efferocytosis function of macrophages, and exert an anti-atherosclerotic effect.

[0065] 1. Animal Experimental Model and Experimental Methods (1) Using ApoE - / - Mice were used as experimental subjects and the experiment was divided into two groups: ApoE - / - Adult male mice + saline intervention group and ApoE - / - Adult male mice were given lithospermic acid orally for 8 weeks to induce atherosclerotic plaque formation in mice.

[0066] (2) The aortic tissue sections were stained with oil red to analyze the size of the plaques. The aortic valves were stained with oil red, H&E, and Masson to analyze the plaque area, necrotic core area, and collagen content in the plaques. The number of Mφ in the plaques, the proportion of pro-inflammatory Mφ and anti-inflammatory Mφ were detected by immunofluorescence. The number of apoptotic bodies in the plaques was detected by TUNEL to analyze the effect of lithospermic acid on the properties of AS plaques.

[0067] (3) Immunohistochemical and immunofluorescent staining were performed on the longitudinal sections of the aorta, the brachiocephalic arteries, and the aortic valve to detect the expression of macrophage common markers MOMA-2 or CD68, M1-specific markers iNOS, TNF-α, and IL-6, M2-specific markers Arg1 and Mrc1, and smooth muscle cell marker α-SMA. The distribution of M1 and M2 macrophages within the plaque was observed.

[0068] 2. In vitro cell test methods (1) Mouse peritoneal macrophages were pretreated with lithospermic acid in vitro. Macrophages were stimulated with oxidized low-density lipoprotein (ox-LDL), lipopolysaccharide (LPS), and interleukin-4 (IL-4), respectively. The mRNA levels of M1-specific markers iNOS, TNF-α, IL-6, and IL1β, and M2-specific markers Fizz1, Arg1, Mrc1, and Ym1 were detected by RT-PCR to analyze the effect of lithospermic acid on the M1 and M2 polarization of macrophages.

[0069] (2) After mouse peritoneal macrophages were pretreated with lithospermic acid in vitro, oxLDL was induced, and then oil red staining was performed to detect the effect of lithospermic acid on macrophage foaming; after mouse peritoneal macrophages were pretreated with lithospermic acid in vitro, they were co-cultured with apoptotic bodies, and flow cytometry analysis was performed to detect the effect of lithospermic acid on macrophage efferocytosis.

[0070] 3. Statistical Analysis All data were statistically processed using SPSS 21.0 software. All experimental results and measurement data were expressed as mean ± standard deviation. Intra-group comparisons were performed using ANOVA and repeated measures analysis of variance, and inter-group comparisons were performed using one-way analysis of variance. P < 0.05 was considered a significant difference.

[0071] 4. Results Analysis (1) Lithospermic acid significantly reduced the area of ​​atherosclerotic plaques in the aorta of atherosclerotic mouse models, increased the area of ​​collagen in the plaques, and enhanced the stability of the plaques.

[0072] Depend on Figure 1It can be seen that compared with the control group (Vehicle), the area of ​​aortic atherosclerotic plaques in the lithospermic acid treatment group (LA) was significantly reduced; Figure 2 It can be seen that compared with the control group (Vehicle), the area of ​​atherosclerotic plaques in the aortic valve region of the lithospermic acid treatment group (LA) was significantly reduced; Figure 3 It can be seen that compared with the control group (Vehicle), the collagen area of ​​atherosclerotic plaques in the lithospermic acid treatment group (LA) was significantly increased.

[0073] (2) Lithospermic acid can induce macrophages in mouse atherosclerotic plaques to polarize toward the anti-inflammatory M2 type, enhance the efferocytosis function of macrophages, and block the progression of atherosclerosis.

[0074] Depend on Figure 4 It can be seen that compared with the control group (Vehicle), the CD68-positive area in the atherosclerotic plaques of the lithospermic acid treatment group (LA) increased significantly, and the CD206-positive area increased significantly, suggesting that macrophages were polarized toward the anti-inflammatory M2 type; Figure 5 It can be seen that compared with the control group (Vehicle), the TUNEL&CD68 co-staining ratio of the lithospermic acid treatment group (LA) was significantly increased, suggesting that the macrophage efferocytosis function was enhanced.

[0075] (3) Based on the results of in vitro cell experiments, it was shown that lithospermic acid can inhibit the M1 inflammatory polarization of macrophages, promote the M2 anti-inflammatory polarization, enhance the efferocytosis function of macrophages, and exert an anti-atherosclerotic effect.

[0076] Depend on Figure 6 It can be seen that compared with the control group (Vehicle), the mRNA levels of IL6, IL1β and TNFα expressed by macrophages in the lithospermic acid treatment group (LA) were significantly reduced, while the mRNA levels of Arg1, Ym1 and Fizz1 were significantly increased, suggesting that lithospermic acid inhibits the M1 inflammatory polarization of macrophages and promotes the M2 anti-inflammatory polarization. Figure 7 It can be seen that compared with the control group (Vehicle), the JURKAT-positive area of ​​macrophages in the lithospermic acid treatment group (LA) was significantly increased, suggesting that the macrophage efferocytosis function was enhanced, exerting an anti-atherosclerotic effect.

[0077] Example 2 This example, based on 16s rRNA gene sequencing results from animal experiments, shows that lithospermic acid treatment significantly alters the composition and abundance of the gut microbiota in a mouse model of atherosclerosis. Fecal microbiota transplantation (FMT) experiments have shown that recipient mice treated with lithospermic acid and then transplanted feces from donor mice showed reduced atherosclerotic plaque size in the aorta and aortic valve, further demonstrating that the gut microbiota plays a key role in the mechanism by which lithospermic acid alleviates atherosclerosis.

[0078] 1. Animal experimental model and experimental methods of lithospermic acid intervention on intestinal flora (1) Using LDLR - / - Mice were used as experimental subjects and the experiment was divided into two groups: LDLR - / - Mice + saline intervention group and LDLR - / - The mice in the lithospermic acid gavage intervention group were fed a high-fat diet for 12 weeks to induce atherosclerotic plaque formation in mice.

[0079] (2) Collect mouse feces and perform 16S ribosomal RNA (16S rRNA) sequencing.

[0080] 2. Animal Experimental Model and Methods of Lithospermic Acid Intervention for Fecal Microbiota Transplantation (1) Using LDLR - / - Mice were used as experimental subjects. All experimental animals were treated with a broad-spectrum antibiotic mixture (Abx) in their drinking water for two weeks and then divided into two groups. One to two days after stopping the antibiotics, the mice were divided into two groups: a donor-0.9% NaCl intervention group (oral gavage of feces for 12 weeks) and a donor-lithospermic acid intervention group (oral gavage of feces for 12 weeks). They were gavaged with fresh fecal homogenate every other day and fed a high-cholesterol diet for 12 weeks to induce atherosclerotic plaque formation.

[0081] (2) The aorta was stained with oil red, and the area of ​​atherosclerotic plaques was analyzed after scanning with a digital film scanner. The mouse hearts were collected, and the aortic valves of the mouse hearts were stained with oil red to analyze the area of ​​atherosclerotic plaques.

[0082] 3. Statistical Analysis All data were statistically processed using SPSS 21.0 software. All experimental results and measurement data were expressed as mean ± standard deviation. Intra-group comparisons were performed using ANOVA and repeated measures analysis of variance, and inter-group comparisons were performed using one-way analysis of variance. P < 0.05 was considered a significant difference.

[0083] 4. Results Analysis (1) After lithospermic acid intervention, the composition and abundance of intestinal flora in the atherosclerosis mouse model showed significant changes.

[0084] Depend on Figure 8 It can be seen that compared with the control group (Vehicle), the composition of the intestinal flora of atherosclerotic mice in the lithospermic acid treatment group (LA) was significantly changed ( Figure 8 A in the genus); In terms of the composition of key species at the genus level, lithospermic acid can increase the abundance of Myxospira and Clostridium, reduce the abundance of Faecalibacterium, and significantly affect the abundance distribution structure of intestinal flora ( Figure 8 B in ).

[0085] (2) The area of ​​atherosclerotic plaques in the aorta and aortic valve of recipient mice was reduced after fecal transplantation of donor mice treated with lithospermic acid. Depend on Figure 9 It can be seen that compared with the fecal transplantation control group (FMT-Control), the fecal transplantation group (FMT-LA) of donor mice treated with lithospermic acid significantly reduced the area of ​​aortic atherosclerotic plaques ( Figure 9 A in the figure); the area of ​​atherosclerotic plaque in the aortic valve region was significantly reduced ( Figure 9 B in Figure 3) The surface intestinal flora is the key component of lithospermic acid's anti-atherosclerotic effect.

[0086] The above description is merely an embodiment of the present application and is not intended to limit the scope of protection of the present application. For those skilled in the art, various modifications and variations of the present application are possible. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present application shall be included in the scope of protection of the present application.

Claims

1. Use of lithospermic acid in the preparation of a drug for treating atherosclerosis. The CAS number of the lithospermic acid is 28831-65-4. The drug exerts an anti-atherosclerotic effect by regulating macrophage polarization, enhancing macrophage efferocytosis function, and remodeling intestinal flora homeostasis.

2. The use according to claim 1, characterized in that: The drug significantly reduces the area of ​​aortic atherosclerotic plaques.

3. The use according to claim 1, characterized in that: The drug significantly increases the collagen area of ​​atherosclerotic plaques and enhances plaque stability.

4. The use according to claim 1, characterized in that: The regulation of macrophage polarization includes inducing polarization of macrophages in atherosclerotic plaques to the anti-inflammatory M2 type.

5. The use according to claim 1, characterized in that: The regulation of macrophage polarization includes inhibiting macrophage M1 inflammatory polarization and promoting M2 anti-inflammatory polarization.

6. The use according to claim 5, characterized in that: The inhibition of macrophage M1 inflammatory polarization is manifested in reducing the mRNA expression levels of IL6, IL1β and TNFα in macrophages; the promotion of M2 anti-inflammatory polarization is manifested in increasing the mRNA expression levels of Arg1, Ym1 and Fizz1 in macrophages.

7. The use according to claim 1, characterized in that: The enhancement of macrophage efferocytosis function is manifested in increasing the ability of macrophages to clear apoptotic bodies.

8. The use according to claim 1, characterized in that: The remodeling of intestinal microbiota homeostasis includes increasing the abundance of Myxospira and Clostridium, and decreasing the abundance of Faecalibacterium.

9. The use according to claim 1, characterized in that: The drug reduces the area of ​​atherosclerotic plaques in the aorta and aortic valve by regulating the homeostasis of intestinal flora.

10. A drug for treating atherosclerosis, characterized in that: The invention comprises lithospermic acid with CAS number 28831-65-4 as claimed in claim 1 and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is selected from a carrier, a diluent or an excipient; and the pharmaceutical dosage form is selected from tablets, granules, capsules, decoctions or injections.

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