Application of cistanche deserticola polysaccharide in preparation of product for preventing and treating heart injury

Abstract

The invention discloses application of cistanche deserticola polysaccharide in preparation of a product for preventing and treating heart injury. The cistanche polysaccharide can significantly improve the survival rate, the left ventricular ejection fraction, the left ventricular shortening fraction, the cardiac output, the myocardial enzyme level of serum and the antioxidant level of doxorubicin-induced cardiotoxicity mice; myocardial cell arrangement disorder, myocardial fiber breakage and myocardial fibrosis caused by adriamycin induction are relieved; the medicine is used for preventing and / or treating doxorubicin-induced heart injury and assisting doxorubicin anti-tumor treatment.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of Cistanche deserticola polysaccharide in the preparation of products for the prevention and treatment of heart damage. Background Technology

[0002] Cistanche deserticola Ma is a perennial parasitic herb belonging to the genus Cistanche in the family Orobanchaceae. It is a precious medicinal plant unique to the arid and desert regions of Northwest my country, with its medicinal history dating back over two thousand years to the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica). It was listed as a "superior" medicinal material, and historical herbal texts (such as the *Compendium of Materia Medica* and *Treatise on the Properties of Medicinal Herbs*) record its effects of "tonifying kidney yang, nourishing essence and blood, and moistening the intestines to relieve constipation." Modern pharmacological research has confirmed that Cistanche deserticola has potential application value in regulating the body's immune function, combating fatigue, protecting nerve cells, and improving reproductive function. The active components of Cistanche deserticola include polysaccharides, phenylethanol glycosides, and iridoids. Among them, Cistanche deserticola polysaccharides are the most abundant and core functional components. In vitro cell experiments (such as cardiomyocyte and hepatocyte models) and animal models (such as mouse and rat models) have confirmed that this polysaccharide exhibits excellent biological activities in scavenging free radicals, inhibiting inflammatory responses, regulating cell metabolism, and protecting organelle structures, providing reliable experimental evidence for its application in the field of drug-induced organ damage protection.

[0003] Doxorubicin (DOX) is a widely used broad-spectrum anthracycline antitumor drug in clinical practice, primarily used to treat solid tumors and hematological malignancies. However, doxorubicin exhibits dose-dependent cardiotoxicity, causing irreversible damage to the heart. This toxicity can be categorized into three types: acute (arrhythmias and pericarditis appearing hours to days after administration), subacute (myocardial damage appearing weeks to months after administration), and chronic (heart failure appearing months to years after administration). Its core pathogenic mechanism is as follows: after entering cardiomyocytes, doxorubicin promotes excessive production of reactive oxygen species (ROS), activates inflammatory signaling pathways, damages mitochondrial structure and function, and induces cardiomyocyte apoptosis, ultimately leading to myocardial fibrosis, ventricular remodeling, and eventually irreversible heart failure. This becomes a key factor limiting the clinical application dosage of doxorubicin and affecting the long-term survival rate of cancer patients. Cardiotoxicity is a major problem that needs to be addressed and resolved in the clinical application of doxorubicin.

[0004] Currently, dexrazoxane is the main drug used clinically to prevent doxorubicin cardiotoxicity. However, it has limitations, including a limited patient population (approved only for breast cancer patients receiving doxorubicin treatment), potential risks of bone marrow suppression, and the possibility of affecting antitumor efficacy when used in combination with chemotherapy drugs. Furthermore, there is no unified clinical consensus on the protective effects of other adjuvant therapies (such as coenzyme Q10 and vitamin E). Therefore, finding doxorubicin cardiotoxicity-protective substances that are naturally derived, highly safe, have a clear mechanism of action, and also possess nutritional regulatory functions has become an important research direction in the field of adjuvant therapy for cancer chemotherapy.

[0005] Existing studies have confirmed the activities of polysaccharides of Cistanche deserticola (CDPS) in antioxidation, anti-inflammation, and organelle protection, but there are no reports on its application in the protection against doxorubicin-induced cardiotoxicity. Summary of the Invention

[0006] To better prevent and treat doxorubicin-induced cardiotoxicity and reduce the side effects of doxorubicin treatment for tumors, this invention provides the following technical solutions.

[0007] In a first aspect, the present invention provides the use of Cistanche deserticola polysaccharide in the preparation of products for the prevention and / or treatment of heart damage.

[0008] Preferably, the application includes at least one of the following: (1) Application in the preparation of products that improve survival rates in cases of cardiac injury; (2) Application in the preparation of products that increase left ventricular ejection fraction, increase left ventricular shortening fraction and / or increase cardiac output in cases of cardiac injury; (3) Application in the preparation of products that reduce cardiomyocyte disorder, myocardial fiber breakage and / or myocardial fibrosis in cases of cardiac injury; (4) Use in the preparation of products that reduce serum creatine kinase levels, serum creatine kinase isoenzyme levels and / or serum lactate dehydrogenase levels in cases of cardiac injury; (5) Application in the preparation of products that increase serum superoxide dismutase levels, decrease serum malondialdehyde levels and / or increase serum glutathione peroxidase levels in cases of cardiac injury.

[0009] Preferably, the cardiac injury is doxorubicin-induced cardiotoxicity.

[0010] Furthermore, the application includes at least one of the following: (1) Application in the preparation of products that improve survival rate; (2) Application in the preparation of products that increase left ventricular ejection fraction, increase left ventricular shortening fraction and / or increase cardiac output; (3) Application in the preparation of products that reduce cardiomyocyte disorder, reduce myocardial fiber breakage and / or reduce myocardial fibrosis; (4) Application in the preparation of products that reduce serum creatine kinase levels, reduce serum creatine kinase isoenzyme levels and / or reduce serum lactate dehydrogenase levels; (5) Application in the preparation of products that increase serum superoxide dismutase levels, decrease serum malondialdehyde levels and / or increase serum glutathione peroxidase levels.

[0011] Preferably, the daily dosage of the Cistanche deserticola polysaccharide is 100-300 mg / kg body weight, for example: 100, 120, 150, 180, 200, 220, 250, 280 or 300 mg / kg body weight.

[0012] Furthermore, the daily dosage of the Cistanche deserticola polysaccharide is 150-250 mg / kg body weight.

[0013] Preferably, the product includes medicine or feed.

[0014] Furthermore, the dosage forms of the drug include tablets, aqueous injections, powder injections, granules, pills, powders, suppositories, emulsions, gels, aerosols, sprays, powder inhalers, capsules, or oral liquids.

[0015] Furthermore, the drug also includes pharmaceutically permissible excipients.

[0016] Furthermore, the excipients include isotonic agents, buffer solutions, flavoring agents, excipients, fillers, binders, disintegrants, or lubricants.

[0017] Preferably, the feed also includes additives that are permitted to be added to the feed.

[0018] Secondly, this invention provides the application of Cistanche deserticola polysaccharide in the preparation of products that assist doxorubicin in antitumor treatment.

[0019] Preferably, the daily dosage of the Cistanche deserticola polysaccharide is 100-300 mg / kg body weight, for example: 100, 120, 150, 180, 200, 220, 250, 280 or 300 mg / kg body weight.

[0020] Furthermore, the daily dosage of the Cistanche deserticola polysaccharide is 150-250 mg / kg body weight.

[0021] Preferably, the tumor includes breast cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, acute lymphoblastic leukemia, acute myeloid leukemia, ovarian cancer, soft tissue sarcoma, osteosarcoma, rhabdomyosarcoma, Ewing's sarcoma, nephroblastoma, neuroblastoma, gastric cancer, pancreatic cancer, liver cancer, prostate cancer, head and neck squamous cell carcinoma, testicular cancer, lung cancer, bladder cancer, or medullary thyroid carcinoma.

[0022] Thirdly, the present invention provides a method for preventing and / or treating doxorubicin-induced cardiotoxicity, comprising administering Cistanche deserticola polysaccharide to a patient.

[0023] Preferably, the daily dosage of the Cistanche deserticola polysaccharide is 100-300 mg / kg body weight, for example: 100, 120, 150, 180, 200, 220, 250, 280 or 300 mg / kg body weight.

[0024] Furthermore, the daily dosage of the Cistanche deserticola polysaccharide is 150-250 mg / kg body weight.

[0025] Fourthly, the present invention provides a method for assisting doxorubicin in antitumor treatment, including the step of administering Cistanche deserticola polysaccharide to a patient.

[0026] Preferably, the daily dosage of the Cistanche deserticola polysaccharide is 100-300 mg / kg body weight, for example: 100, 120, 150, 180, 200, 220, 250, 280 or 300 mg / kg body weight.

[0027] Furthermore, the daily dosage of the Cistanche deserticola polysaccharide is 150-250 mg / kg body weight.

[0028] The beneficial effects of this invention are: This invention, through systematic animal experiments, demonstrates that Cistanche deserticola polysaccharides can significantly improve the survival rate, left ventricular ejection fraction, left ventricular shortening fraction, cardiac output, serum myocardial enzyme levels, and antioxidant levels in mice with doxorubicin-induced cardiotoxicity; it also alleviates doxorubicin-induced cardiomyocyte disorder, myocardial fiber rupture, and myocardial fibrosis; Cistanche deserticola polysaccharides can be used to prevent and / or treat doxorubicin-induced cardiotoxicity. This invention provides a new technical solution for developing novel doxorubicin-induced cardiotoxicity protective products and for assisting doxorubicin in antitumor treatment, possessing extremely high clinical application value and social significance. Attached Figure Description

[0029] Figure 1 The diagram shown is a schematic of the animal experiment process; Figure 2 The figure shows the survival curves of the experimental mice; Figure 3The images show echocardiograms and related statistical charts of experimental mice; A shows B-mode and M-mode echocardiograms; B shows a bar chart of left ventricular ejection fraction (LVEF); C shows a bar chart of left ventricular fractional shortening (LVFS); and D shows a bar chart of cardiac output (CO). Figure 4 The image shown is a HE staining image of a cardiac pathological section from an experimental mouse. Figure 5 The image shown is a Sirius red staining image of a cardiac pathological section from an experimental mouse. Figure 6 The table shows the serum myocardial enzyme levels of experimental mice; A represents the creatine kinase (Ck) level, B represents the creatine kinase isoenzyme (Ck-mb) level, and C represents the lactate dehydrogenase (Ldh) level. Figure 7 The table shows the serum antioxidant levels of experimental mice; A represents the superoxide dismutase (SOD) level, B represents the malondialdehyde (MDA) level, and C represents the serum glutathione peroxidase (GSH-PX) level. Detailed Implementation

[0030] The technical solution of the present invention will be further described below with reference to embodiments and accompanying drawings. The advantages and features of the present invention will become clearer as the description unfolds. However, it should be understood that the embodiments are merely exemplary and do not constitute a limitation on the scope of the present invention.

[0031] It should be noted that, unless otherwise specified, the experimental methods used in the following embodiments are conventional methods in the art. Unless otherwise defined, all scientific and technical terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art.

[0032] Example 1: Effects of Cistanche deserticola polysaccharide on doxorubicin-induced cardiotoxicity model mice 1.1 Animal Experiments according to Figure 1 The experimental procedures for mice were followed. All C57BL / 6J mice were housed in a pathogen-free facility with access to standard mouse feed and water, and subjected to a 12-hour light / 12-hour dark cycle under controlled temperature conditions. The mouse experiments were conducted in accordance with guidelines for the use and care of laboratory animals and were approved by the Ethics Committee of China Agricultural University. Specific experiments are as follows: Forty-two male 8-week-old C57BL / 6J mice were randomly divided into a control group, a model group, and an intervention group, with 14 mice in each group. The model and intervention groups received intraperitoneal injections of doxorubicin (DOX) at a dose of 3 mg / kg body weight every other day, for a cumulative dose of 21 mg / kg, to induce cardiotoxicity. Mice in the intervention group received gavage with Cistanche deserticola polysaccharide (CDPs) solution at a dose of 200 mg / kg body weight / day. Mice in the control group received the same amount of saline as the intervention group via gavage, and mice in the model group received the same amount of DOX via intraperitoneal injection with saline. For mice that died during the experiment, the heart was dissected on the same day, preserved in tissue fixative, and sectioned for HE and Sirius red staining. For the remaining mice, echocardiography was performed the day after the last administration, and relevant cardiac parameters were recorded. The eyeballs were then removed, and blood was collected. The blood was centrifuged at 3500 rpm for 5 min, and serum was used to detect myocardial enzyme and antioxidant levels.

[0033] 1.2 Detection of experimental mouse indicators 1.2.1 After the mice in each group were finally administered the drug, the survival rate of each group was calculated.

[0034] like Figure 2 As shown, the survival rate of the model group mice was significantly lower than that of the control group mice, indicating that the model was successfully established. The survival rate of the intervention group mice was significantly higher than that of the model group mice, and there was no significant difference in survival rate between the intervention group and the control group. This indicates that Cistanche deserticola polysaccharide can significantly improve doxorubicin-induced cardiotoxicity in mice and can be used for the prevention and / or treatment of doxorubicin-induced cardiotoxicity.

[0035] 1.2.2 Echocardiography was used to detect left ventricular ejection fraction, left ventricular shortening fraction, and cardiac output, and statistical analysis was performed to determine whether there were significant differences among the groups.

[0036] 1.2.3 The differences in cardiac pathology among the three groups of mice were observed by staining heart sections with HE and Sirius red.

[0037] 1.2.3.1 Preparation of cardiac slices The left ventricle of each mouse heart was perfused with 5 mL of 0.01M PBS (pH=7.4), allowing unrestricted outflow from the vena cava incision. The mouse heart was carefully dissected under a microscope to remove as much excess tissue as possible, and then the heart was immersed in 4% paraformaldehyde fixative for at least 24 h.

[0038] The heart samples preserved in the fixative were embedded according to the embedding steps in Table 1.

[0039] Table 1. Embedding process of cardiac tissue After embedding the heart tissue, it was left to stand at room temperature for 1 day before sectioning. The heart paraffin block was cut into 5 μm paraffin sections, which were then dried overnight in a 37°C oven for subsequent staining. Before staining, the paraffin sections needed to be dewaxed according to the steps in Table 2.

[0040] Table 2 Dewaxing process of paraffin sections The dewaxed tissue sections can be used for subsequent staining procedures.

[0041] 1.2.3.2 Hematoxylin and eosin (HE) staining of heart sections ① Stain with hematoxylin solution for 3 minutes, then wash away excess dye with distilled water.

[0042] ② Differentiate in the differentiation solution for 30 seconds, then rinse twice with tap water for 2 minutes each time.

[0043] ③ Place in eosin staining solution for 30 seconds, then wash away excess stain with distilled water. Do not leave for too long.

[0044] ④ Dehydrate in 75% ethanol, 85% ethanol, 95% ethanol and 100% ethanol in sequence, and rinse each level quickly for 2-3 seconds to prevent discoloration.

[0045] ⑤ Place the solution in xylene I and xylene II in sequence until it becomes transparent, and rinse each step for 8 minutes.

[0046] ⑥ Mount the slide with neutral resin and observe it under an optical microscope after it has dried completely.

[0047] 1.2.3.3 Modified Sirius Red staining for heart sections: ① Mix equal amounts of iron hematoxylin stock solution (A1) and iron hematoxylin dilution solution (A2) and use immediately after preparation.

[0048] ② Stain with the prepared iron hematoxylin staining solution for 5 minutes, then wash with distilled water for 15 seconds to remove excess staining solution.

[0049] ③ Soak in tap water for 5 minutes, then rinse with distilled water 3 times, 10 seconds each time.

[0050] ④ Stain with Sirius red staining solution for 5 minutes, rinse slightly with running water to remove the stain from the surface of the slide. ⑤ Dehydrate in 75% ethanol, 85% ethanol, 95% ethanol and 100% ethanol in sequence, and rinse each level quickly for 2-3 seconds to prevent discoloration.

[0051] ⑥ Mount the slide with neutral resin and observe it under an optical microscope after it has dried completely.

[0052] 1.2.4 Serum Myocardial Enzyme Level Detection: Mouse blood samples were placed in coagulation tubes and incubated at room temperature for 2-3 hours. The samples were then centrifuged at 3000 rpm for 15 minutes, and the supernatant pale yellow serum was collected for later use. Serum myocardial enzyme levels were detected using a creatine kinase (Ck) assay kit (colorimetric method) (Nanjing Jiancheng Bioengineering Institute, catalog number A032-1-1), a creatine kinase isoenzyme (Ck-mb) assay kit (enzyme-linked immunosorbent assay) (Nanjing Jiancheng Bioengineering Institute, catalog number H197-1-1), and a lactate dehydrogenase (Ldh) assay kit (microplate method, upgraded version) (Nanjing Jiancheng Bioengineering Institute, catalog number A020-4-1). The statistical analysis was performed to determine if there were significant differences between the data from each group.

[0053] 1.2.5 Serum Antioxidant Level Detection: Mouse blood samples were placed in coagulation tubes and incubated at room temperature for 2-3 hours. The samples were then centrifuged at 3000 rpm for 15 minutes, and the supernatant pale yellow serum was collected for later use. Serum superoxide dismutase (T-SOD) levels were detected using a total superoxide dismutase (T-SOD) assay kit (WST-1 method) (Nanjing Jiancheng Bioengineering Institute, A001-3-2). Malondialdehyde (MDA) levels were detected using a malondialdehyde (MDA) assay kit (TBA method) (Nanjing Jiancheng Bioengineering Institute, A003-1-2). Glutathione peroxidase (GSH-PX) levels were detected using a glutathione peroxidase (GSH-PX) assay kit (Nanjing Jiancheng Bioengineering Institute, A005-1-2) (colorimetric method). The statistical analysis was performed to determine if there were significant differences between the data from each group.

[0054] The data processing was performed using Graphpad Prism 10.0 software. Statistical results are expressed as mean ± standard deviation (±SD). When comparing the three groups of test data, one-way ANOVA was used, and p < 0.05 was considered statistically significant. * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001.

[0055] like Figure 3 As shown in (AD), compared to the control group mice, the model group mice showed significantly lower left ventricular ejection fraction, left ventricular shortening fraction, and cardiac output, indicating successful model establishment. Compared to the model group mice, the intervention group mice showed significantly increased left ventricular ejection fraction, left ventricular shortening fraction, and cardiac output. However, there was no significant difference between the intervention group and the control group mice. This indicates that Cistanche deserticola polysaccharide can significantly improve doxorubicin-induced cardiotoxicity in mice and can be used for the prevention and / or treatment of doxorubicin-induced cardiotoxicity.

[0056] like Figure 4-5As shown, compared with the control group mice, the model group mice exhibited significantly more disordered cardiomyocyte arrangement, myocardial fiber rupture, and myocardial fibrosis symptoms, indicating successful model establishment. Compared with the model group mice, the intervention group mice showed significantly reduced symptoms of disordered cardiomyocyte arrangement, myocardial fiber rupture, and myocardial fibrosis. This demonstrates that Cistanche deserticola polysaccharide can significantly improve doxorubicin-induced cardiotoxicity in mice and can be used for the prevention and / or treatment of doxorubicin-induced cardiotoxicity.

[0057] like Figure 6 As shown, compared with the control group mice, the serum levels of creatine kinase (Ck), creatine kinase isoenzyme (Ck-mb), and lactate dehydrogenase (Ldh) in the model group mice were significantly increased, indicating that the model was successfully established. Compared with the model group mice, the serum levels of Ck, Ck-mb, and Ldh in the intervention group mice were significantly decreased, but there was no significant difference between them and the control group mice. This indicates that Cistanche deserticola polysaccharide can significantly improve doxorubicin-induced cardiotoxicity in mice and can be used for the prevention and / or treatment of doxorubicin-induced cardiotoxicity.

[0058] like Figure 7 As shown, compared with the control group mice, the serum SOD level in the model group mice was significantly decreased, the MDA level was significantly increased, and the GSH-PX level was significantly decreased, indicating that the model was successfully established. Compared with the model group mice, the serum SOD level in the intervention group mice was significantly increased, the MDA level was significantly decreased, and the GSH-PX level was significantly increased, but there was no significant difference between the intervention group and the control group mice. This indicates that Cistanche deserticola polysaccharide can significantly improve doxorubicin-induced cardiotoxicity in mice and can be used for the prevention and / or treatment of doxorubicin-induced cardiotoxicity.

[0059] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

Claims

1. Application of Cistanche deserticola polysaccharide in the preparation of products for the prevention and / or treatment of heart damage.

2. The application according to claim 1, characterized in that, The application includes at least one of the following: (1) Application in the preparation of products that improve survival rates in cases of cardiac injury; (2) Application in the preparation of products that increase left ventricular ejection fraction, increase left ventricular shortening fraction and / or increase cardiac output in cases of cardiac injury; (3) Application in the preparation of products that reduce cardiomyocyte disorder, reduce myocardial fiber breakage and / or reduce myocardial fibrosis in the case of cardiac injury; (4) Application in the preparation of products that reduce serum creatine kinase levels, reduce serum creatine kinase isoenzyme levels and / or reduce serum lactate dehydrogenase levels in cases of cardiac injury; (5) Application in the preparation of products that increase serum superoxide dismutase levels, decrease serum malondialdehyde levels and / or increase serum glutathione peroxidase levels in cases of cardiac injury.

3. The application according to claim 1, characterized in that, The cardiac injury is doxorubicin-induced cardiotoxicity; and / or, the daily dosage of the Cistanche deserticola polysaccharide is 100-300 mg / kg body weight.

4. The application according to claim 1, characterized in that, The product is either a medicine or feed.

5. The application according to claim 4, characterized in that, The dosage forms of the drug include tablets, aqueous injections, powder injections, granules, pills, powders, suppositories, emulsions, gels, aerosols, sprays, powder inhalers, capsules, or oral liquids.

6. The application according to claim 4, characterized in that, The drug also includes pharmaceutically permissible excipients.

7. The application according to claim 6, characterized in that, The excipients include isotonic agents, buffer solutions, flavoring agents, excipients, fillers, binders, disintegrants, or lubricants.

8. The application according to claim 4, characterized in that, The feed also includes any additives that are permitted to be added to the feed.

9. Application of Cistanche deserticola polysaccharide in the preparation of products that assist doxorubicin in antitumor treatment.

10. The application according to claim 9, characterized in that, The tumors include breast cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, acute lymphoblastic leukemia, acute myeloid leukemia, ovarian cancer, soft tissue sarcoma, osteosarcoma, rhabdomyosarcoma, Ewing's sarcoma, nephroblastoma, neuroblastoma, gastric cancer, pancreatic cancer, liver cancer, prostate cancer, squamous cell carcinoma of the head and neck, testicular cancer, lung cancer, bladder cancer, or medullary thyroid carcinoma.

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