Application of schisandrin B in preparation of medicine for preventing or treating intestinal and kidney injury caused by ionizing radiation
By verifying the antioxidant and anti-inflammatory functions of Schisandra ethinin, a mouse model was used to explore its protective effect on intestinal and kidney damage caused by ionizing radiation, and a drug with Schisandra ethinin as the main component was prepared, which solved the toxicity of existing drugs. The problem of long half-life significantly alleviates intestinal and kidney damage caused by radiation therapy complications and provides a new direction for the development of radiation protection drugs.
Patent Information
- Application Number
- CN202411414369.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-11
- Publication Date
- 2025-01-17
AI Technical Summary
Existing drugs to prevent and treat radiation therapy complications, such as amifostin, have problems with large toxicity and long half-life, which leads to their limitations in clinical use. It is urgent to develop related radiation protection drugs, which are caused by Schisandra Ethrin for ionizing radiation. The potential protective effect of related tissue damage has not been reported.
By verifying the antioxidant, anti-stress and anti-inflammatory functions of Schisandra ethinin, the systemic and abdominal irradiation model of mice was used to explore the protective effect of Schisandra ethinin on intestinal and kidney damage caused by ionizing radiation, and the preparation was mainly made. Ingredients for preventing or treating intestinal and kidney damage caused by ionizing radiation.
Schisandra et al. significantly prolongs the survival time of mice, improves survival rates, reduces intestinal and kidney damage, has radiation protection effects, and provides a potential drug development direction to treat radioactive intestinal and kidney damage.
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Figure CN119318651A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the field of medicine, in particular to the application of schisandrin B, an effective ingredient in schisandra chinensis, in the preparation of medicines for preventing or treating intestinal and kidney damage caused by ionizing radiation. Background Art
[0002] With the development of clinical technology, radiotherapy (RT) is widely used in cancer patients. About 60% of cancer patients receive radiotherapy to improve survival rate. During the whole treatment process, healthy tissues in the radiation area will inevitably be damaged. Radiation-induced intestinal injury (RIII) is a common complication of radiotherapy. RIII is mainly manifested by necrosis and sloughing of small intestinal mucosal epithelium, apoptosis and necrosis of crypt cells, reduction of crypt number, mucosal inflammation and intestinal flora ectopic. In addition, the kidneys close to the pelvic cavity are also sensitive organs in radiotherapy. Radiation nephropathy (RN) is one of the important radiation toxicities during radiotherapy for gastrointestinal and gynecological cancers, upper abdominal sarcomas or lymphomas. It is reported that victims of acute radiation accidents and more than 20% of patients who receive total body irradiation (TBI) before hematopoietic stem cell transplantation will develop RN. Currently, most drugs for preventing and treating RT complications are in the research stage. Amifostine (WR-2721), the only anti-radiation drug approved by the FDA, has problems such as high toxicity and long half-life, which greatly limits its clinical use. There is an urgent need to develop related radiation protection drugs.
[0003] Schisandrin B (Schisandrin B) is the benzocyclooctene lignan with the highest content in the traditional Chinese medicine Schisandra chinensis. Studies have shown that it has the effects of preventing inflammation, oxidative stress and tumor cell proliferation. However, the potential protective effect of Schisandrin B on related tissue damage caused by ionizing radiation has not been reported so far. Summary of the invention
[0004] The present invention aims to provide a new use of schisandrin B in preparing a drug for preventing or treating intestinal and kidney damage caused by ionizing radiation.
[0005] In order to achieve the above object, the present invention adopts the following technical solution:
[0006] Schisandrin B, as one of the important active ingredients of Schisandrin, has been verified to have biological functions such as antioxidant, anti-stress, and anti-inflammatory. The present invention established a moderate to severe acute radiation sickness model by whole-body irradiation of mice to explore the radiation protection effect of Schisandrin B on mice, and then focused on the protective effect of Schisandrin B on acute intestinal and renal damage caused by abdominal irradiation in mice.
[0007] Based on the above technical scheme, the first aspect of the present invention provides the use of schisandrin B in the preparation of drugs for preventing or treating intestinal and kidney damage caused by ionizing radiation.
[0008] Furthermore, the drug for preventing or treating intestinal and kidney damage caused by ionizing radiation is a drug composition composed of schisandrin B as the only active monomer component, or schisandrin B and at least one pharmaceutically acceptable excipient.
[0009] Furthermore, the purity of schisandrin B in the drug for preventing or treating intestinal and kidney damage caused by ionizing radiation is not less than 99.9%.
[0010] Furthermore, the dosage of schisandrin B in the drug for preventing or treating intestinal and kidney damage caused by ionizing radiation is 20 mg / kg.
[0011] Furthermore, the drug for preventing or treating intestinal and kidney damage caused by ionizing radiation is an oral preparation or an intravenous injection.
[0012] Furthermore, the site of intestinal damage caused by ionizing radiation is selected from at least one of the small intestine, colon, and rectum. The site of renal damage caused by ionizing radiation is selected from the renal tubule-related area.
[0013] Furthermore, the radiation in the ionizing radiation is selected from gamma rays.
[0014] Furthermore, the drug for preventing or treating intestinal and kidney damage caused by ionizing radiation is a drug that reduces intestinal and kidney pathological damage, maintains the stability of the intestinal mucosal barrier, and reduces the content of kidney injury markers.
[0015] Furthermore, the drug for preventing or treating intestinal and kidney damage caused by ionizing radiation is a drug that prolongs survival time after irradiation, significantly improves survival rate, and reduces intestinal damage after whole-body irradiation.
[0016] Furthermore, the drug for preventing or treating intestinal and kidney damage caused by ionizing radiation is a drug for alleviating acute kidney damage caused by radiation.
[0017] The second aspect of the present invention provides a drug for preventing or treating intestinal and kidney damage caused by ionizing radiation. The drug for preventing or treating intestinal and kidney damage caused by ionizing radiation contains schisandrin B as the only active monomer ingredient, or schisandrin B and at least one pharmaceutically acceptable excipient to form a pharmaceutical composition.
[0018] The advantages of the present invention are:
[0019] The present invention provides the use of schisandrin B in preventing or treating intestinal and renal damage caused by ionizing radiation. A moderate to severe acute radiation sickness model is established by whole-body irradiation of mice, and the radiation protection effect of schisandrin B on mice is explored. The results show that schisandrin B can significantly prolong the survival time of mice after whole-body irradiation and improve the survival rate of mice. Further studies show that schisandrin B can significantly alleviate intestinal damage, and then focus on the protective effect of acute intestinal and renal damage caused by abdominal irradiation. A mouse acute intestinal and renal damage model is established by local abdominal irradiation of mice. The results show that schisandrin B can significantly alleviate radiation-induced intestinal and renal damage in mice, indicating that it has a certain radiation protection effect. Therefore, schisandrin B has a wide range of application prospects in the treatment of radiation intestinal and renal damage and drug development as a potential radiation protector. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] Figure 1 This is a schematic diagram of the protective effect of schisandrin B on whole-body irradiation in mice.
[0021] Figure 2 This is a schematic diagram of the protective effect of Schisandra chinensis on intestinal damage induced by abdominal irradiation in mice.
[0022] Figure 3 This is a schematic diagram of the protective effect of Schisandra chinensis on kidney damage induced by abdominal irradiation in mice. DETAILED DESCRIPTION
[0023] The specific implementation methods provided by the present invention are described in detail below in conjunction with examples.
[0024] The schisandrae B used in the experiment in the embodiment of the present invention was purchased from (MCE China). The SPF-grade male C57BL / 6J mice used in the experiment were 6-8 weeks old and weighed about 20±3g, purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd. (Beijing, China). The room temperature was about 20-25°C, and they were free to drink distilled water and eat standard mouse food. The experimental process and related operations were approved by the Experimental Animal Center of the Academy of Military Medical Sciences.
[0025] Example 1: Study on the protective effect of schisandrin B on whole body irradiation in mice
[0026] 1. Experimental Grouping and Drug Administration
[0027] Animal experiment: divided into normal group, irradiation group (IR), irradiation + Schisandrin B (IR + Sch B). Normal group: mice were injected with 200 μL normal saline in the abdomen every day; irradiation group: normal saline was injected intraperitoneally 12h before irradiation, 2h, 12h, 24h after irradiation, and whole-body γ-ray (8.5Gy) irradiation; irradiation + Schisandrin B group: Sch B was injected intraperitoneally 12h before irradiation, 2h, 12h, 24h after irradiation, and whole-body γ-ray (8.5Gy) irradiation.
[0028] 2. Experimental Methods
[0029] (1) Monitoring of the survival rate of mice in each group
[0030] After each group was treated accordingly, the mortality rate of mice in each group was checked every day and recorded in real time, and the survival rate of mice in each group was counted within 30 days.
[0031] (2) Evaluation of mouse excretion status / colon length statistics
[0032] After the corresponding treatment, the colon length of each group of mice was counted and photographed for comparison. The excretion (feces volume) of each group of mice was counted for three consecutive days and photographed for comparison.
[0033] (3) Hematoxylin-eosin staining to detect intestinal tissue pathological damage
[0034] The tissue sections were immersed in xylene I and II for 30 minutes respectively; then immersed in different concentrations of ethanol in turn, and then rinsed with distilled water for 1 minute; then stained with hematoxylin-eosin; the stained slides were sealed with neutral resin and observed under an optical microscope.
[0035] (4) Immunohistochemical staining of intestinal tissue
[0036] 1) The intestinal tissue was fixed with 4% paraformaldehyde for two days, then replaced with 70% ethanol to preserve the tissue, and then the tissue was trimmed into 5 mm tissue blocks and placed in an embedding box;
[0037] 2) The tissue was dehydrated by soaking in 70% ethanol, 85% ethanol, 90% ethanol, 95% ethanol and anhydrous ethanol for 60 min, then treated with dewaxing solution I, dewaxing solution II and dewaxing solution III for 30 min, and then treated with paraffin I, paraffin II and paraffin III for 60 min;
[0038] 3) After the tissue was embedded with an embedding machine, the paraffin sections were cut into 3 μm, and then the paraffin sections were dewaxed. The process was as follows: baking (60°C, 120 min) → soaking in dewaxing solution for 5 min (xylene II, xylene I, alcohol + xylene, 100% alcohol II, 100% alcohol I, 95% alcohol, 90% alcohol, 85% alcohol, 70% alcohol) → washing with distilled water twice;
[0039] 4) Antigen retrieval was performed using EDTA (pH 9.0) at 121°C (treatment for 2 min), and the sections were washed twice with PBS after natural cooling, and then blocked with hydrogen peroxide and 1% BSA in sequence (washed twice with PBS in between), and then incubated with primary antibody (prepared with 1% BSA) at 4°C overnight;
[0040] 5) After incubation with primary antibody overnight, the primary antibody was discarded and the slides were washed three times with PBS. Then, the slides were incubated with horseradish peroxidase-labeled secondary antibody for 90 min and washed three times with PBS. Then, the slides were developed with 3-diaminobenzidine (DAB), counterstained with hematoxylin, and washed to blue. Finally, the slides were dehydrated and sealed with neutral gum.
[0041] 3. Experimental results
[0042] like Figure 1 As shown, Figure 1 This is a schematic diagram of the results of the study on the radiation protection effect of Schisandrin B. Figure 1 It can be seen that Figure 1 A and B show that Schisandrin B can significantly prolong the survival time of mice after irradiation, significantly improve the survival rate of mice, and reduce the intestinal damage of mice after whole-body irradiation ( Figure 1 C,D).
[0043] Example 2: Study on the protective effect of Schisandra chinensis on abdominal irradiation intestinal damage in mice
[0044] 1. Experimental Grouping and Drug Administration
[0045] Animal experiment: divided into normal group, irradiation group (IR), irradiation + Schisandrin B (IR + Sch B). Normal group: mice were injected with 200 μL of normal saline in the abdomen every day; irradiation group: normal saline was injected intraperitoneally 12h before irradiation, 2h, 12h, and 24h after irradiation, and the abdomen was irradiated with γ-rays (13Gy); irradiation + Schisandrin B group: Schisandrin was injected intraperitoneally 12h before irradiation, 2h, 12h, and 24h after irradiation, and the abdomen was irradiated with γ-rays (13Gy).
[0046] 2. Experimental Methods
[0047] (1) Statistics of colon length of mice in each group
[0048] After each group was treated accordingly, the colon length of each group of mice was counted and photographed for comparison.
[0049] (2) Hematoxylin-eosin staining to detect intestinal tissue pathological damage
[0050] The method is the same as the detection method in Example 1.
[0051] (3) Immunofluorescence staining of intestinal tissue
[0052] The method is the same as the detection method in Example 1.
[0053] 3. Experimental results
[0054] like Figure 2 As shown, Figure 2 This is a schematic diagram of the protective effect of schisandrin B on intestinal damage induced by abdominal irradiation in mice.
[0055] from Figure 2 It can be seen that Schisandrin B can significantly reduce the colon atrophy of mice after abdominal irradiation ( Figure 2 A, F), reducing intestinal tissue damage after abdominal irradiation in mice ( Figure 2 B, C), the intestinal mucosal barrier is an important defense against the invasion of pathogenic bacteria, and irradiation greatly damages the intestinal mucosal barrier. ZO-1 and Occludin are two intestinal tight junction proteins. Immunofluorescence staining of ZO-1 and Occludin was used to detect the effect of Schisandrin B on the intestinal mucosal barrier. The results showed that the administration of Schisandrin B could significantly improve the intestinal barrier damage after abdominal irradiation in mice, and had a certain protective effect on radiation-induced intestinal damage ( Figure 2 D, E).
[0056] Example 3: Study on the protective effect of schisandrin B on kidney damage caused by abdominal irradiation in mice
[0057] 1. Experimental Grouping and Drug Administration
[0058] The specific method is the same as the grouping and administration scheme in Example 2
[0059] 2. Experimental Methods
[0060] (1) Hematoxylin-eosin staining to detect pathological damage of renal tissue
[0061] The method is the same as the detection method in Example 1.
[0062] (2) ELISA was used to determine the levels of related injury indicators in the kidney tissues of mice in each group
[0063] 1) Add 100 μl of standard samples with different gradient concentrations and samples to be tested into the ELISA plate in duplicate.
[0064] 2) Seal the ELISA plate with a sealing film and place it in a 37°C incubator for 90 minutes.
[0065] 3) After the reaction is completed, the ELISA plate is dried, and then 100 μl of the corresponding indicator antibody working solution is added to each well in turn. The pipette is repeatedly blown to mix, and the plate is sealed with a sealing film and incubated at 37°C for 60 minutes.
[0066] 4) After the reaction is completed, pour out the reagents in the wells of the ELISA plate, wash five times by adding 300 μl of washing buffer to each well, and then tap out the washing solution.
[0067] 5) After washing the ELISA plate, use a pipette to add 90 μl of TMB colorimetric solution to each well, and then incubate in a 37°C incubator in the dark for 20 minutes.
[0068] 6) After the reaction, take out the ELISA plate and add 100 μl of the stop solution to each well using a pipette. Immediately after adding, place the plate in an ELISA reader for detection and measure its absorbance at 450 nm.
[0069] 2. Experimental results
[0070] like Figure 3 As shown, Figure 3 This is a schematic diagram of the protective effect of schisandrin B on renal damage induced by abdominal irradiation in mice.
[0071] from Figure 3 As can be seen, the HE staining results showed that compared with the model group, Schisandrin B could significantly reduce the renal tissue damage caused by abdominal irradiation in mice ( Figure 3 A), modern research shows that neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) are one of the most effective biological markers for diagnosing acute kidney injury. They are usually expressed at very low concentrations in human tissues, but after acute kidney injury, NGAL and KIM-1 levels increase rapidly, which has important clinical value in the diagnosis of acute kidney injury. When kidneys are injured, serum creatinine, as a traditional biological marker, will also increase rapidly. The content of these three key kidney injury biomarkers was detected by enzyme-linked immunosorbent assay. The results showed that Schisandrin B can significantly reduce radiation-induced acute kidney injury ( Figure 3 BD).
[0072] The preferred embodiments of the present invention have been specifically described above, but the present invention is not limited to the embodiments. Those skilled in the art may make various equivalent modifications or substitutions without violating the spirit of the present invention. These equivalent modifications or substitutions are all included in the scope defined by the claims of this application.
Claims
1. Application of schisandrin B in the preparation of drugs for preventing or treating intestinal and kidney damage caused by ionizing radiation.
2. The use of Schisandrin B according to claim 1 in the preparation of a drug for preventing or treating intestinal and kidney damage caused by ionizing radiation, characterized in that: The drug for preventing or treating intestinal and kidney damage caused by ionizing radiation is a drug composition composed of schisandrin B as the only active monomer component, or schisandrin B and at least one pharmaceutically permitted auxiliary material.
3. The use of Schisandrin B according to claim 2 in the preparation of a drug for preventing or treating intestinal and kidney damage caused by ionizing radiation, characterized in that: The purity of schisandrin B in the medicine for preventing or treating intestinal and kidney damage caused by ionizing radiation is not less than 99.9%.
4. The use of Schisandrin B according to claim 2 in the preparation of a drug for preventing or treating intestinal and kidney damage caused by ionizing radiation, characterized in that: The dosage of schisandrin B in the medicine for preventing or treating intestinal and kidney damage caused by ionizing radiation is 20 mg / kg.
5. The use of Schisandrin B according to claim 2 in the preparation of a drug for preventing or treating intestinal and kidney damage caused by ionizing radiation, characterized in that: The medicine for preventing or treating intestinal and kidney damage caused by ionizing radiation is an oral preparation or an intravenous injection.
6. The use of Schisandrin B according to claim 1 in the preparation of a drug for preventing or treating intestinal and kidney damage caused by ionizing radiation, characterized in that: The site of intestinal damage caused by ionizing radiation is selected from at least one of the small intestine, colon, and rectum; the site of renal damage caused by ionizing radiation is selected from the renal tubule-related area.
7. The use of Schisandrin B according to claim 1 in the preparation of a drug for preventing or treating intestinal and kidney damage caused by ionizing radiation, characterized in that: The radiation in the ionizing radiation is selected from gamma rays.
8. The use of Schisandrin B according to claim 1 in the preparation of a drug for preventing or treating intestinal and kidney damage caused by ionizing radiation, characterized in that: The drug for preventing or treating intestinal and kidney damage caused by ionizing radiation is a drug that reduces intestinal and kidney pathological damage, maintains the stability of the intestinal mucosal barrier, and reduces the content of kidney damage markers.
9. The use of Schisandrin B according to claim 1 in the preparation of a drug for preventing or treating intestinal and kidney damage caused by ionizing radiation, characterized in that: The drug for preventing or treating intestinal and kidney damage caused by ionizing radiation is a drug that prolongs the survival time after irradiation, significantly improves the survival rate, reduces intestinal damage after whole-body irradiation, and reduces acute kidney damage caused by radiation.
10. A drug for preventing or treating intestinal and kidney damage caused by ionizing radiation, characterized in that: The drug for preventing or treating intestinal and kidney damage caused by ionizing radiation uses schisandrin B as the only active monomer component, or schisandrin B and at least one pharmaceutically permitted auxiliary material to form a drug composition.