Application of schisandra chinensis in prevention and treatment of cardiovascular diseases caused by microplastics

Abstract

This invention discloses the application of Schisandra chinensis in the prevention and treatment of cardiovascular diseases caused by microplastics. The schisandrin B described in this invention is consistent with the "low-dose, long-term" exposure characteristics of microplastics, emphasizing prevention and early intervention, which helps to control the development of cumulative damage. By targeting and inhibiting the sustained activation of aryl hydrocarbon receptors (AhR), it intervenes in the initiation of microplastic-specific toxic pathways at the molecular level, which is expected to more effectively block the chronic damage process and is more fundamental and targeted than general antioxidant / anti-inflammatory strategies that only target downstream symptoms.

Description

Technical Field

[0001] This invention relates to the technical field of cardiovascular damage caused by microplastics, and specifically to the application of Schisandra chinensis in the prevention and treatment of cardiovascular diseases caused by microplastics. Background Technology

[0002] With the globalization of plastic pollution, microplastics smaller than 5 millimeters and even smaller nanoplastics have become emerging environmental pollutants. They can enter the human body through various pathways, including drinking water, food, and air, and accumulate in tissues and organs. The cardiovascular system is one of their key targets, and epidemiological and toxicological studies both indicate a close correlation between microplastic exposure and increased cardiovascular disease risk. Unlike traditional cardiovascular risk factors (such as hypertension and hyperlipidemia), microplastics / nanoplastics, as exogenous foreign particles, exhibit a unique time- and dose-dependent pattern of cardiovascular toxicity: long-term, low-dose chronic exposure is the primary scenario for health hazards, and the damaging effects gradually appear and intensify with prolonged exposure time and increased cumulative dose in the body. This unique exposure-effect relationship suggests that their pathogenic mechanism may be fundamentally different from endogenous metabolic disorders. Therefore, traditional cardiovascular prevention and treatment strategies are not entirely applicable or have limited effectiveness, and there is an urgent need to develop intervention methods targeting the characteristics of these emerging pollutants.

[0003] Currently, prevention and treatment of cardiovascular damage caused by microplastics mainly focus on environmental interventions to reduce microplastic exposure and the use of drugs to treat the symptoms of cardiovascular damage. In terms of environmental interventions, optimizing the manufacturing process of plastic products and strengthening wastewater treatment to remove microplastics reduces the risk of human exposure. However, these methods cannot address the issue of internal damage repair in already exposed individuals. Cardiovascular disease prevention and treatment drugs (such as statins, ACE inhibitors, and antioxidant vitamins) are primarily designed to target endogenous metabolic disorders or known pathological processes, rather than the persistent cellular stress and specific receptor pathway activation caused by exogenous particulate matter. While these drugs may alleviate some downstream symptoms caused by microplastic exposure (such as oxidative stress and inflammation) to some extent, they fail to address the unique toxic pathways initiated by microplastics as a physicochemical complex. Therefore, their effectiveness in preventing and addressing the root causes of progressive and cumulative cardiovascular damage caused by long-term microplastic accumulation is insufficient.

[0004] Schisandra chinensis, a traditional Chinese medicine and food, has been used for over two thousand years. The pharmacological effects of its active components (such as schisandrin A, schisandrin B, and schisandrol A) have been extensively studied. Current technology confirms that it possesses various activities including antioxidant, anti-inflammatory, hepatoprotective, neuroprotective, and cardiovascular protective properties, and can be used to assist in improving myocardial ischemia and regulating blood lipids in common cardiovascular diseases. However, current technology does not address the application of Schisandra chinensis and its active components in addressing cardiovascular damage induced by long-term, low-dose exposure to microplastics / nanoplastics, nor has it been reported to inhibit specific signaling pathways such as the aryl hydrocarbon receptor (AhR) that are continuously activated under such exposure. Summary of the Invention

[0005] In view of this, the present invention provides an application of Schisandra chinensis in the prevention and treatment of cardiovascular diseases caused by microplastics. The present invention clarifies and utilizes the effect of Schisandra chinensis and its active ingredients in inhibiting the sustained activation of AhR caused by microplastics, thereby making up for the shortcomings of traditional prevention and treatment methods in the treatment of related diseases caused by microplastics.

[0006] The specific technical solution of the present invention is as follows: 1. The application of Schisandra chinensis in the preparation of drugs for the prevention and treatment of cardiovascular diseases caused by microplastics.

[0007] 2. Application of schisandrin B in the preparation of drugs for the prevention and treatment of cardiovascular diseases caused by microplastics.

[0008] Furthermore, the microplastics refer to plastic particles with a size ≤ 5 mm.

[0009] Furthermore, cardiovascular diseases refer to a class of diseases in which abnormalities occur in the structure or function of the heart and blood vessels, leading to obstruction of blood flow.

[0010] Furthermore, the cardiovascular diseases include those that cause damage to the myocardium.

[0011] Furthermore, the drug exerts its effect by acting on the aryl hydrocarbon receptor signaling pathway.

[0012] Furthermore, the drug dosage form may be an injection, tablet, capsule, granule, or decoction.

[0013] 3. The application of Schisandra chinensis or Schisandrin B in the preparation of health food products, which have an auxiliary protective effect against cardiovascular damage caused by microplastics.

[0014] 4. The application of Schisandra chinensis or schisandrin B in the preparation of functional foods, which have health benefits against cardiovascular damage caused by microplastics.

[0015] The beneficial effects of this invention are as follows: To address the shortcomings of existing cardiovascular prevention and treatment programs in intervening in the specific and cumulative cardiovascular damage caused by long-term, low-dose exposure to microplastics / nanoplastics, this invention provides a solution applicable to long-term intervention and targeting core pathological pathways activated by microplastics / nanoplastics (such as the AhR pathway). Specifically, it utilizes Schisandra chinensis and its active ingredients in the preparation of products for preventing and treating cardiovascular damage caused by microplastics / nanoplastics, addressing their unique exposure patterns and toxic mechanisms. This invention investigates the emerging cardiovascular risk of long-term environmental exposure to microplastics / nanoplastics, filling a gap in existing cardiovascular prevention and treatment systems in this area. Unlike traditional drugs that focus on treating established diseases, the Schisandrin B described in this invention aligns with the "low-dose, long-term" exposure characteristics of microplastics, emphasizing prevention and early intervention. This helps control the development of cumulative damage. By targeting and inhibiting the sustained activation of the aryl hydrocarbon receptor (AhR), it intervenes at the molecular level initiating the specific toxic pathways of microplastics, potentially more effectively blocking the chronic damage process. This approach is more fundamental and targeted than general antioxidant / anti-inflammatory strategies that only target downstream symptoms.

[0016] Other advantages, objectives, and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination, or may be learned from practice of the invention. The objectives and other advantages of the invention can be realized and obtained through the following description. Attached Figure Description

[0017] To make the objectives, technical solutions, and advantages of the present invention clearer, the preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings, wherein: Figure 1 To observe the uptake of PS NPs in mouse myocardial tissue using transmission electron microscopy (scale bar = 1µm or 500nm), red arrows indicate PS NP particles.

[0018] Figure 2 Echocardiographic results of mice with myocardial injury induced by polystyrene nanoparticles (PS NPs) for different durations; Figure 3 Masson staining results of mouse myocardial tissue induced by polystyrene nanoparticles (PS NPs) for different durations of myocardial injury; Figure 4 The results show the mRNA expression of Anp, Bnp, and Myh7 in mouse myocardial tissue induced by polystyrene nanoparticles (PS NPs) for different durations of myocardial injury. Figure 5 The results of AhR mRNA expression in the heart tissue of mice exposed to PS NPs for 28 and 42 days; Figure 6The results of AhR protein expression in the heart tissue of mice exposed to PS NPs for 28 and 42 days; Figure 7 Results of AhR nucleocytoplasmic ratio in heart tissues of mice exposed to PS NPs for 28 and 42 days; Figure 8 The results of mRNA expression of Anp, Bnp, and Myh7 in H9c2 cells stimulated with different doses of polystyrene nanoparticles (PS NPs) for 48 h. Figure 9 Masson staining results of myocardial tissue in mice to alleviate PS NPs-induced myocardial injury using schisandrin B (SchB); Figure 10 Results of mRNA expression of Anp, Bnp, and Myh7 in myocardial tissue of mice to alleviate PS NPs-induced myocardial injury by schisandrin B (SchB); Figure 11 The results show the effect of Sch B on the mRNA expression level of AhR in the heart tissue of PS NPs-exposed mice; Figure 12 The results show the effect of Sch B on the expression level of AhR protein in the heart tissue of PS NPs-exposed mice; Figure 13 The results of the nucleocytoplasmic ratio for the effect of Sch B on AhR activity in the heart tissue of PS NPs-exposed mice; Figure 14 Results of schisandrin B (SchB) reducing the mRNA expression of Anp, Bnp, and Myh7 in PS NPs-stimulated H9c2 cells. Detailed Implementation

[0019] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention. It should also be noted that technical means not described in detail in this invention can be implemented using conventional technical means.

[0020] I. Deposition Experiment of Polystyrene Nanoparticles (PS NPs) in Cardiomyocytes C57BL / 6 mice were treated with PS NPs 75 mg / kg by gavage. Fresh heart tissue was harvested 12 hours later and cut into 1 mm pieces. 3Tissue blocks were initially fixed with 2.5% glutaraldehyde at 4°C. After thorough washing with phosphate-buffered saline (PBS), the samples were post-fixed in 1% osmium tetroxide for 1 hour, followed by washing with PBS again. The tissues were then subjected to a series of ethanol dehydration treatments, incubated with pure acetone for 20 minutes, impregnated with a 1:1 mixture of epoxy resin and acetone for 3 hours, and finally embedded in pure epoxy resin overnight. Ultrathin sections were prepared using an ultramicrotome, stained with uranium acetate and lead citrate sequentially, and observed using a transmission electron microscope (JEM-1400PLUS, NEC Corporation, Tokyo, Japan). The results are as follows: Figure 1 As shown, PS NPs are deposited in cardiomyocytes, mainly distributed in the cytoplasm, indicating that PS NPs can be taken up by cardiomyocytes.

[0021] II. Time-dependent experiment on myocardial injury induced by polystyrene nanoparticles (PS NPs) C57BL / 6 mice were randomly divided into three groups: (1) a solvent control group, (2) a PS NPs treatment group for 28 days, and (3) a PSNPs treatment group for 42 days. The solvent control group received 0.3 mL of Milli-Q water by gavage daily, while the PS NPs group received 200 µg / (mL·day) of PS NPs by gavage. Echocardiographic results are shown in […]. Figure 2 Compared with the control group, mice exposed to PS NPs for 28 or 42 days had significantly reduced ejection fraction (EF) and fractional shortening (FS) (p<0.05 or p<0.01), indicating myocardial contractile dysfunction.

[0022] Masson staining results of mouse myocardial tissue are shown in the figure. Figure 3 After statistical analysis, the software revealed that, compared with the control group, the myocardial fibrosis area of ​​mice exposed for 28 days increased by about 0.6%, and that of mice exposed for 42 days increased by about 5.4% (p<0.01).

[0023] The mRNA expression results of Anp, Bnp, and Myh7 in mouse myocardial tissue are shown in the figure. Figure 4 Real-time quantitative PCR (RT-PCR) analysis showed that the mRNA expression of myocardial injury-related genes (Anp, Bnp and Myh7) was significantly upregulated after 28 and 42 days of PS NP exposure (p<0.01).

[0024] Aromatic hydrocarbon receptors (AhRs) were initially considered targets for mediating the toxicity of environmental pollutants such as polycyclic aromatic hydrocarbons (PAHs), but they have now been transformed into important physiological regulators of immune responses. Figure 5 The experimental results showed that the expression level of AhR mRNA in the heart tissue of mice exposed to PS NPs for 42 days was significantly increased compared with that of mice exposed for 28 days (p<0.01).

[0025] Figure 6 The experimental results showed that the expression level of AhR protein in the heart tissue of mice exposed to PS NPs for 42 days was significantly increased compared with that of mice exposed for 28 days (p<0.01).

[0026] Figure 7 Immunofluorescence staining results showed that the nucleocytoplasmic ratio of AhR in cardiac tissue was significantly increased (p<0.05 or p<0.01), indicating enhanced AhR activation.

[0027] III. Dose-dependent experiments on myocardial injury induced by polystyrene nanoparticles (PS NPs) In vitro experiments were conducted by stimulating H9c2 cells with different concentrations (100, 200 µg / mL) of PS NPs for 48 hours, while other cells were cultured using standard cell culture methods. The expression results of Anp, Bnp, and Myh7 mRNA in H9c2 cells are shown in the table below. Figure 8 The results showed that both doses of PS NPs significantly upregulated Anp mRNA expression, while the 200 µg / mL dose simultaneously increased Bnp and Myh7 mRNA expression (p<0.05 or p<0.01).

[0028] IV. Schisandrin B (SchB) in vitro and in vivo experiments on the reduction of PSNP-induced myocardial injury. C57BL / 6 mice were randomly divided into 4 groups, with 5 mice in each group. The control group was given 0.3 mL of 0.5% CMC-Na (carboxymethyl cellulose sodium) solution daily; the PS NPs group was given a mixture of 75 mg / kg PS NPs and 0.5% CMC-Na; the 30 mg / kg SchB group was given 75 mg / kg PS NPs and 30 mg / kg SchB; and the 60 mg / kg SchB group was given 75 mg / kg PS NPs and 60 mg / kg SchB. The experiment lasted for 42 days. Masson staining results are shown in the figure. Figure 9 Software statistical analysis revealed that PSNPs could induce myocardial interstitial fibrosis, with the fibrin-positive area in the PSNP group increasing by approximately 6% compared to the control group (p<0.01); Sch B treatment could reduce the fibrin-positive area in the PSNP group, with a reduction of approximately 2% in the 30 mg / kg treatment group and approximately 4% in the 60 mg / kg treatment group (p<0.01).

[0029] The mRNA expression levels of myocardial injury-related genes (Anp, Bnp, Myh7) in PS NPs-exposed mice are shown in the figure. Figure 10The results showed that the expression levels of each gene were significantly upregulated (p<0.01), while Sch B treatment significantly downregulated the expression levels of these genes (p<0.01).

[0030] The effect of Sch B on AhR mRNA expression levels in the heart tissue of PS NPs-exposed mice is shown in the figure. Figure 11 The results showed that the expression levels of AhR mRNA in the myocardial tissue of mice in the 30 and 60 mg / kg Sch B groups were significantly reduced (p<0.01).

[0031] Figure 12 This indicates that the expression level of AhR protein in the myocardial tissue of mice in the 30 and 60 mg / kg Sch B groups was significantly reduced (p<0.01).

[0032] Figure 13 The nucleocytoplasmic ratio results showed that AhR activity was significantly reduced in the myocardial tissue of mice in the 30 and 60 mg / kg Sch B groups (p<0.01).

[0033] Figure 11-13 The results all indicate that AhR is a potential target for Sch B in treating PS NPs-induced myocardial injury.

[0034] The specific procedures for the in vitro experiment are as follows: H9c2 rat cardiomyocytes were cultured in high-glucose DMEM medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin, and were routinely cultured in a 37 ℃, 5% CO2 incubator. When the cell confluence reached 70%–80%, the medium was replaced with serum-free medium for synchronization for 12 h. Subsequently, the cells were divided into a control group, PS NPs (200 μg / mL), and Sch B treatment groups (200 μg / mL PS NPs and 1, 3 and 10 µM Sch B), and cultured for 48 hours.

[0035] In vitro experimental results as follows Figure 14 As shown, in H9c2 cells, Sch B (1, 3 and 10 µM) significantly downregulated the expression of myocardial injury-related genes (Anp, Bnp and Myh7) in the PSNPs-exposed group (p<0.05 or p<0.01).

[0036] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. The application of Schisandra chinensis in the preparation of drugs for the prevention and treatment of cardiovascular diseases caused by microplastics.

2. Application of schisandrin B in the preparation of drugs for the prevention and treatment of cardiovascular diseases caused by microplastics.

3. The application according to claim 1 or 2, characterized in that, The microplastics refer to plastic particles with a size ≤ 5 mm.

4. The application according to claim 1 or 2, characterized in that, Cardiovascular diseases refer to a class of diseases caused by abnormalities in the structure or function of the heart and blood vessels, resulting in obstruction of blood flow.

5. The application according to claim 1 or 2, characterized in that, The cardiovascular diseases mentioned include those that cause damage to the myocardium.

6. The application according to claim 1 or 2, characterized in that, The drug works by acting on the aryl hydrocarbon receptor signaling pathway.

7. The application according to claim 1 or 2, characterized in that, The drug dosage form may be an injection, tablet, capsule, granule, or decoction.

8. The application of Schisandra chinensis or schisandrin B in the preparation of health food products, characterized in that, Health foods have a protective effect against cardiovascular damage caused by microplastics.

9. The application of Schisandra chinensis or schisandrin B in the preparation of functional foods, characterized in that, Functional foods have health benefits against cardiovascular damage caused by microplastics.

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