Application of hemp seed oil in the preparation of drugs for the prevention or mitigation of acute ionizing radiation-induced diseases.

Abstract

This invention discloses the application of hemp seed oil in the preparation of drugs for the prevention or mitigation of acute ionizing radiation-induced diseases, relating to the field of biomedical technology. This invention discovers that hemp seed oil can protect the intestinal epithelium and crypt structures under acute high-dose irradiation, maintaining the intestinal barrier and digestive function. Simultaneously, it can protect splenic immune function by slowing the reduction in the white pulp region of the spleen and increasing the number of nucleated cells and lymphoid immune cells in the spleen, thereby significantly improving the individual survival rate after lethal doses of radiation, providing a new approach for treating acute ionizing radiation-induced diseases.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, and more specifically, to the use of hemp seed oil in the preparation of remedies for the prevention or mitigation of acute ionizing radiation-induced diseases. Background Technology

[0002] In specific production accidents and extreme wartime environments, workers and the general public may face the risk of acute ionizing radiation. High-dose ionizing radiation causes water molecules in cells to ionize, producing excessive reactive oxygen species (ROS), putting cells into a state of oxidative stress. Simultaneously, it induces DNA breaks, activates downstream pathways such as p38 and p53, leading to cellular senescence, apoptosis, and abnormal proliferation. As the radiation dose increases, organs such as the bone marrow, intestines, and brain suffer severe damage in sequence, leading to abnormalities in hematopoietic, immune, digestive, and cognitive functions, and potentially causing death within a short period. Therefore, developing drugs that can effectively prevent and treat high-dose ionizing radiation damage is crucial for improving survival time after acute high-dose radiation exposure and enhancing public awareness of radiation exposure risks.

[0003] Currently, various chemical drugs have been proven to have radiation damage prevention and treatment effects, including estrogens, thiol-based drugs, and antioxidants. These drugs mainly exert their radiation protection effects by reducing ROS levels, inhibiting inflammatory signals, and promoting cell proliferation. However, there are many limitations in terms of their types and applications. For example, WR-2721, which is currently the most widely accepted, not only causes serious side effects such as hypotension and dizziness, but also requires injection within half an hour before radiation exposure, and the concentration of its secondary effective metabolites in tissues drops rapidly within half an hour, resulting in a very limited application window.

[0004] In comparison, natural ingredient-based drugs have the advantages of high safety and long-lasting efficacy. Reported natural radiation-protective drug components include flavonoids, polysaccharides, and polyphenols. Flavonoids and polyphenols achieve their anti-oxidative stress effects by providing phenolic hydroxyl radicals to reduce free radicals, while polysaccharides possess multiple pharmacological effects, including immunomodulation, antioxidation, and anti-inflammation. These natural ingredient-based drugs have lower toxicity and side effects than chemical drugs; however, the efficacy evidence for their radiation-protective effects in animal experiments needs further refinement, and clinical application has not yet been realized.

[0005] In view of this, the present invention is proposed. Summary of the Invention

[0006] The purpose of this invention is to provide the application of hemp seed oil in the preparation of drugs for the prevention or mitigation of acute ionizing radiation-induced diseases.

[0007] This invention is implemented as follows:

[0008] In a first aspect, the present invention provides the use of hemp seed oil in the preparation of drugs for the prevention or mitigation of acute ionizing radiation-induced diseases.

[0009] In an optional implementation, the radiation dose of the acute ionizing radiation injury is greater than the half-lethal radiation dose.

[0010] In an optional embodiment, the dose rate of the acute ionizing radiation injury is 0.5-1 Gy / min.

[0011] In an optional embodiment, the radiation source of the acute ionizing radiation injury includes a gamma radiation source;

[0012] Preferably, the gamma radiation source includes at least one of 60Co and 137Cs.

[0013] In an optional embodiment, the hemp seed oil is administered by injection or oral administration.

[0014] Preferably, the hemp seed oil is administered 0.1-12 hours before radiation.

[0015] In an alternative implementation, the prevention or mitigation of acute ionizing radiation-induced diseases is achieved by improving post-irradiation survival rates.

[0016] In an optional implementation, improving survival rate after irradiation is achieved by protecting the intestinal epithelium and crypt structures, maintaining the intestinal barrier and digestive function.

[0017] In an alternative implementation, improving post-irradiation survival is achieved by protecting the spleen's immune function;

[0018] Preferably, the protection of spleen immune function is achieved by increasing the number of nucleated cells and lymphoid immune cells in the spleen;

[0019] Preferably, the protection of spleen immune function is achieved by slowing down the reduction of the white pulp region of the spleen.

[0020] In a second aspect, the present invention provides the use of a pharmaceutical composition in the preparation of a medicament for preventing or mitigating acute ionizing radiation injury to improve survival rate, said pharmaceutical composition comprising hemp seed oil.

[0021] In an optional embodiment, the pharmaceutical composition may further include one or more pharmaceutically acceptable excipients.

[0022] The present invention has the following beneficial effects:

[0023] This application proposes the use of hemp seed oil in the preparation of drugs for the prevention or mitigation of acute ionizing radiation-induced diseases. This expands the drug selection for high-dose radiation injuries. Furthermore, hemp seed oil is a natural drug with few side effects. It can protect the integrity of the intestinal epithelial barrier and crypt structure, preventing infections caused by acute radiation injury. It can also, to some extent, avoid lymphocyte depletion caused by lethal doses of radiation, thereby significantly improving individual survival rates after lethal doses of radiation. Pharmaceutical compositions containing this hemp seed oil also possess similar technical effects. Attached Figure Description

[0024] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0025] Figure 1 This is a schematic diagram showing the changes in the 30-day survival rate of different groups of mice after 8.5 Gy radiation, as provided in Example 1 of this application.

[0026] Figure 2 This is a schematic diagram showing the changes in body weight of different groups of mice after 30 days following 8.5 Gy radiation, as provided in Example 1 of this application.

[0027] Figure 3 HE staining images of multiple tissues 5 days after 8.5 Gy radiation provided in Example 2 of this application;

[0028] Figure 4 A statistical chart of the number of nucleated cells in the spleen on day 5 after 8.5 Gy radiation, provided in Example 3 of this application;

[0029] Figure 5 A statistical chart showing the percentage of spleen lymphocytes on day 5 after 8.5 Gy radiation, provided in Example 3 of this application. Detailed Implementation

[0030] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0031] Hemp seeds (Fructus cannabis), also known as hemp seeds, white hemp seeds, or hemp kernels, are the dried, mature seeds of the artificially cultivated cannabis plant (Cannabis sativa L.) belonging to the Moraceae family. They were first mentioned in the *Daily Materia Medica* and *Shennong's Classic of Materia Medica*, and their medicinal use was first recorded in the *Shennong's Classic of Materia Medica*. They are an important medicinal and edible herb in traditional Chinese medicine. Hemp seed oil is one of the most common processed products of hemp seeds. It contains a large amount of unsaturated fatty acids such as oleic acid, linoleic acid, and α-linolenic acid, and has effects such as lowering blood lipids, anti-oxidation, and improving memory. It is widely used as a typical medicinal and edible herb and a health-promoting food.

[0032] Previous studies by Hu Xuefeng et al., who fed rats 10 mL / kg of hemp seed oil for 90 days, found that compared with the soybean oil group and the control group, the rats' serum levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were significantly increased, while the lipid peroxidation product malondialdehyde (MDA) was significantly decreased, suggesting that hemp seed oil has a significant effect on enhancing the body's antioxidant capacity. Su Jing et al. treated D-galactose-induced aging mice with hemp seed oil and found that the mice's learning and memory abilities were significantly improved, which may be related to hemp seed oil's ability to enhance the brain tissue's antioxidant and free radical scavenging capabilities and strengthen the central cholinergic nervous system. In the same model mice, after administration of 6 and 12 mL / kg of hemp seed oil for 42 days, the serum levels of C-reactive protein, tumor necrosis factor α, and Toll receptor 4 were significantly reduced, indicating that hemp seed oil also has an anti-inflammatory effect. However, to date, there are no studies on the radiation protection effects of hemp seed oil.

[0033] Therefore, this application provides a new use for hemp seed oil. The research in this application found that hemp seed oil has potential application value in the preparation of drugs for the prevention or mitigation of acute ionizing radiation-induced diseases.

[0034] It should be noted that the acute ionizing radiation referred to in this application differs from conventional solar radiation or electromagnetic radiation. Electromagnetic radiation typically causes sunburn and is generally safe for humans. Ionizing radiation, however, exhibits a degree of randomness at low doses, meaning the effects are uncertain when the radiation dose received is small. High doses, on the other hand, have a deterministic effect; when the radiation dose received is substantial, corresponding radiation damage will occur. This application addresses acute ionizing radiation damage resulting from high radiation doses.

[0035] Specifically, in this application, the radiation dose for acute ionizing radiation injury is greater than the half-lethal radiation dose. The dose rate for acute ionizing radiation injury is 0.5-1 Gy / min.

[0036] The term "half-lethal radiation dose" refers to the absorbed dose required to kill 50% of a biological population within a specified time. For acute whole-body irradiation, the mortality rate is about 50% when the human body receives a dose equivalent of 450 rem. If the dose equivalent exceeds 600 rem, the mortality rate is almost 100%.

[0037] The acute effects of radiation refer to the effects that occur when a person receives a large dose of radiation in a single instance or over a short period of time. For the human body, radiation exceeding 1 Gy causes a rapid decrease in white blood cells in the blood, weakened immunity, and impaired self-renewal capacity of hematopoietic stem cells in the bone marrow; radiation exceeding 5 Gy causes gastrointestinal damage; radiation exceeding 8 Gy causes severe heart damage and cardiovascular disease; and radiation exceeding 20 Gy causes severe damage to the brain and central nervous system.

[0038] Radiation sources causing acute ionizing radiation injury include gamma radiation sources; preferably, gamma radiation sources include at least one of 60Co and 137Cs. It should be understood that the hemp seed oil provided in this application can also be used for protection against other similar radiation sources.

[0039] Hemp seed oil can be administered by injection or oral administration; preferably, it is administered 0.1-12 hours before radiation. Better results can be achieved by administering the oil before radiation, and in practice, multiple administrations can be given before and after irradiation to enhance the protective effect.

[0040] In this application, the prevention or mitigation of acute ionizing radiation-induced diseases is achieved by increasing the survival rate after irradiation.

[0041] Improving post-irradiation survival rate can be achieved by protecting the intestinal epithelium and crypt structures, maintaining the intestinal barrier and digestive function; or by protecting the spleen's immune function, which is achieved by increasing the number of nucleated cells and lymphoid immune cells in the spleen; and by slowing down the reduction in the white pulp region of the spleen.

[0042] Furthermore, this invention provides the use of a pharmaceutical composition in the preparation of a medicament for preventing or mitigating acute ionizing radiation injury to improve survival rates, the pharmaceutical composition comprising hemp seed oil. The pharmaceutical composition also includes one or more pharmaceutically acceptable excipients.

[0043] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0044] The hemp seed oil used in this application embodiment is a commercially available edible hemp seed oil, branded as Daoxinyuan, and its product name is: Daoxinyuan Bama Hemp Seed Oil.

[0045] Example 1: Verification that hemp seed oil can improve the survival rate after irradiation.

[0046] Forty-five mice were divided into three groups (unirradiated group Ctrl, irradiated group IR, and irradiated group + hemp seed oil administration group IR+HSO), with 15 mice in each group.

[0047] In both the irradiation group (IR) and the irradiation + hemp seed oil administration group (IR+HSO), a single irradiation at a cobalt-60 radiation source lasting 12 min 9 s was performed, with a dose rate of 0.7 Gy / min and a total dose of 8.5 Gy. The irradiation group (IR) received no medication, while the irradiation + hemp seed oil administration group (IR+HSO) received a single dose of 7.5 mL per kilogram of body weight, with hemp seed oil injected 12 hours and 0.5 hours before irradiation, and 12 hours and 24 hours after irradiation.

[0048] The survival rate and body weight of mice 30 days after irradiation were statistically analyzed. (See attached statistical results). Figure 1 and Figure 2 .

[0049] from Figure 1 and Figure 2 It can be seen that, compared with IR, the survival rate of mice in the IR+HSO group increased from 0 to over 60% after 30 days of irradiation, and the weight of surviving mice remained at a stable level, which is better than that of the irradiation group (IR), demonstrating a very good acute high-dose radiation protection effect.

[0050] Example 2: Verification that hemp seed oil can reduce radiation damage to multiple organs.

[0051] Sixty mice were divided into four groups (unirradiated group Ctrl, irradiated group IR, irradiated group + amifostine group IR + Amifostine, and irradiated group + hemp seed oil group IR + HSO), with 15 mice in each group.

[0052] In the irradiation group (IR), the irradiation + amifostine administration group (IR + Amifostine), and the irradiation + hemp seed oil administration group (IR + HSO), all underwent a single 12 min 9 s irradiation from a cobalt-60 radiation source at a dose rate of 0.7 Gy / min, with a total dose of 8.5 Gy. The irradiation group (IR) received no medication. In the irradiation + amifostine administration group, the single dose of amifostine was 150 mg per kilogram of body weight, administered as a single injection 0.5 hours before irradiation. In the irradiation hemp seed oil administration group, the single dose of hemp seed oil was 7.5 mL per kilogram of body weight, administered as injections 12 hours and 0.5 hours before irradiation, and 12 hours and 24 hours after irradiation.

[0053] HE staining was performed on multiple tissues of mice 5 days after irradiation. The specific procedure is as follows:

[0054] Multiple tissues, including the brain, spleen, intestines, and bone marrow, were fixed, dehydrated, embedded, sectioned, stained with hematoxylin and eosin (HE), destained, and mounted. They were then observed under a microscope for analysis of radiation-sensitive tissues.

[0055] Please see Figure 3 Bone marrow analysis showed that, compared to the unirradiated group (Control), the irradiated group (IR) and the irradiated + hemp seed oil treatment group (IR+HSO) had virtually no nucleated cells in their bone marrow, with red blood cells being the main component. The irradiated + amifostine treatment group (IR+Amifostine) also showed a lack of nucleated cells in its bone marrow, with obvious gaps.

[0056] Analysis of the intestines showed that, compared with the unirradiated group (Control), the villi in the irradiated group (IR) were significantly shortened and damaged, and the crypt structure was not obvious. In contrast, compared with the unirradiated group (Control), the irradiated group (IR+HSO) showed no significant changes in villi length and epithelial integrity, and the crypt structure was still clearly discernible. The morphology of intestinal epithelial cells in the irradiated group (IR+Amifostine) was not significantly different from both the unirradiated group (Control) and the irradiated group (IR+HSO).

[0057] Analysis of the spleen showed that the white pulp region of the spleen was significantly reduced in the irradiation group (IR), with a decrease in the number of germinal centers and marginal zones, and indistinct boundaries. In contrast, the white pulp region of the spleen in the irradiation + hemp seed oil administration group (IR+HSO) was reduced, but the internal structures and boundaries were still clearly distinguishable, and the area was significantly larger than that in the IR group. Meanwhile, the irradiation + amifostine administration group and the irradiation + hemp seed oil administration group (IR+HSO) showed similar effects, with the white pulp region still clearly distinguishable, and the area reduced but significantly larger than that in the IR group.

[0058] Analysis of the brain showed no significant changes in brain structure between groups, suggesting that the 8.5 Gy irradiation dose did not cause structural changes in the brain.

[0059] Therefore, HSO can protect the intestinal epithelium and crypt structures during lethal dose irradiation, maintain the intestinal barrier and digestive function, and has a protective effect on the crypt structures where small intestinal stem cells are located; at the same time, it can protect the integrity of immune response-related structures in the spleen, suggesting that it has a protective effect against acute radiation injury to the spleen's immune function. Therefore, hemp seed oil can significantly reduce radiation damage to multiple organs.

[0060] Example 3: Verification that hemp seed oil can have a radioprotective effect on T cell-mediated spleen immune function.

[0061] Forty-five mice were divided into three groups (unirradiated group Ctrl, irradiated group IR, and irradiated group + hemp seed oil administration group IR+HSO), with 15 mice in each group.

[0062] In both the irradiation group (IR) and the irradiation + hemp seed oil administration group (IR + HSO), a single irradiation at a cobalt-60 radiation source lasting 12 min 9 s was performed, with a dose rate of 0.7 Gy / min and a total dose of 8.5 Gy. The irradiation group (IR) received no medication, while the irradiation group (IR + HSO) received a single dose of 7.5 mL per kilogram of body weight, administered 12 hours and 0.5 hours before irradiation, and 12 hours and 24 hours after irradiation.

[0063] Five days after irradiation, cell counts and flow cytometry phenotypic analysis were performed on the spleens of the mice.

[0064] Cell count results showed that, from Figure 4 It can be seen that the number of nucleated cells in the IR group decreased to an extremely low level. In contrast, the number of nucleated cells in the spleen of the IR+HSO group increased significantly, further suggesting that hemp seed oil has a radioprotective effect on the spleen.

[0065] Flow cytometry phenotyping analysis showed that, from Figure 5 It can be seen that in nucleated cells, CD8 in the IR group + T cell and B cell counts were significantly lower in the IR+HSO group compared to the Control group, while CD4 counts were significantly lower in the IR+HSO group. + T cells and CD8 + The proportion of T cells was significantly increased compared to the IR group, indicating that hemp seed oil has a radioprotective effect on T cell-mediated spleen immune function.

[0066] In summary, repeated administration of hemp seed oil can protect the integrity of the intestinal mucosal barrier and the number of immune cells under acute high-dose radiation conditions, thereby significantly improving individual survival rates. This application proposes the use of hemp seed oil in the preparation of drugs for the prevention or mitigation of acute ionizing radiation-induced diseases, which can expand the drug selection for high-dose radiation injury. Furthermore, hemp seed oil is a natural drug with few side effects. It can protect the integrity of the intestinal epithelial barrier and crypt structure, prevent infection caused by acute radiation injury, and can also, to some extent, avoid lymphocyte depletion caused by lethal dose radiation, thus significantly improving individual survival rates after lethal dose radiation. Pharmaceutical compositions containing this hemp seed oil also have similar technical effects.

[0067] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. The use of hemp seed oil in the preparation of medicines for the prevention or mitigation of acute ionizing radiation-induced diseases, characterized in that, The radiation dose of the acute ionizing radiation injury is greater than the half-lethal radiation dose. Preventing or mitigating the aforementioned acute ionizing radiation-induced diseases is achieved by improving post-irradiation survival rates, which is achieved by protecting the intestinal epithelium and crypt structures, maintaining the intestinal barrier and digestive function, or by protecting the spleen's immune function.

2. The application according to claim 1, characterized in that, The dose rate of the acute ionizing radiation injury is 0.5-1 Gy / min.

3. The application according to claim 1, characterized in that, The radiation sources of the acute ionizing radiation injury include gamma radiation sources.

4. The application according to claim 3, characterized in that, The gamma radiation source includes at least one of 60Co and 137Cs.

5. The application according to claim 1, characterized in that, The hemp seed oil is administered by injection or orally.

6. The application according to claim 5, characterized in that, The hemp seed oil was administered 0.1-12 hours before radiation.

7. The use of a pharmaceutical composition in the preparation of a medicament for preventing or mitigating acute ionizing radiation injury to improve survival rate, characterized in that, The pharmaceutical composition is made from hemp seed oil, and the radiation dose of the acute ionizing radiation injury is greater than the half-lethal radiation dose.

8. The application according to claim 7, characterized in that, The pharmaceutical composition also includes one or more pharmaceutically acceptable excipients.

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