CN115089626A - Application of dogwood in preparation of anti-radiation medicine - Google Patents

Abstract

The invention discloses an application of dogwood in preparing a medicine for treating or preventing ionizing radiation damage, in particular an application in preparing a medicine for preventing/treating radioactive bone marrow damage caused by ionizing radiation. The dogwood has a protection effect on natural ionizing radiation and artificial radiation, can be used as a medicament for protecting living body damage caused by radioactive substance radiation, nuclear power station leakage, space ionizing radiation and tumor radiotherapy, and solves the current situation that radioprotection/treatment medicaments are in short supply.

Description

Application of dogwood in preparation of anti-radiation medicine

Technical Field

The invention belongs to the technical field of medicines, and relates to application of dogwood in preparation of medicines for treating or preventing ionizing radiation damage, in particular to application of dogwood in preparation of medicines for preventing/treating radioactive bone marrow damage caused by ionizing radiation. The dogwood has a protective effect on natural ionizing radiation and artificial radiation, and can be used as a medicine for protecting living body injury caused by radioactive substance radiation, nuclear substance leakage, space ionizing radiation and tumor radiotherapy.

Background

The following discussion is presented in the context of a description of the invention to aid in understanding the invention, but is not to be construed as prior art to the invention. All cited publications are incorporated herein by reference in their entirety.

In daily life, radiation is ubiquitous and can be divided into non-ionizing radiation and ionizing radiation.

Non-ionizing radiation refers to radiation that is relatively low in energy and does not ionize material atoms or molecules. Non-ionizing radiation includes low energy electromagnetic radiation, such as ultraviolet, light, infrared, microwave, and radio waves. Their energy is not high, but only causes the particles in the material to vibrate and the temperature to rise.

Ionizing radiation has stronger energy than non-electric radiation and has greater influence on human body. Ionizing radiation includes natural ionizing radiation, artificial radiation. Natural ionizing radiation includes cosmic rays, earth's native radionuclides, decay products of radon in the air, and various naturally occurring radionuclides; the artificial radiation includes medical diagnostic X-ray, radioactive dust from atmospheric nuclear weapons test, radioactive waste discharged from nuclear industry, industrial gamma-ray, ⁶⁰ Co rays, heavy ion radiation, radiation in radiotherapy treatment of tumors and the like. In recent years, the nuclear energy utilization of human beings is deepened (nuclear power plants, medical diagnosis and the like), and the human beings are exposed to more ionizing radiation pollution risks.

Currently, there are very limited drugs for the treatment of ionizing radiation damage, and only about 11 radiation prevention/treatment drugs are on the market worldwide. Based on the risk, relevant researches are carried out, and the research on numerous traditional Chinese medicines and natural medicines for a long time shows that the dogwood extract has an obvious radiation protection effect.

Cornus officinalis (Cornus officinalis sieb. et Zucc.) is mainly produced in shanxi, gansu and shandong of China. The fruit is named as cornus pulp, commonly named as jujube peel, is used for medicine, has sour and astringent taste and mild property, has the efficacies of tonifying liver and kidney and controlling nocturnal emission and controlling depletion, and is a common traditional Chinese medicine with a formula for regulating immunity.

The Corni fructus contains Corni fructus glycoside, Corni fructus neo-glycoside, ursolic acid, gallic acid, malic acid, vitamins, tannin and saponin, and has antibacterial, tranquilizing, pathogenic wind expelling, blood pressure lowering, immunity improving, and cancer cell diffusion inhibiting effects. Modern medical research shows that the dogwood has the function of resisting the AIDS virus.

The dogwood can adopt dogwood medicinal materials and powder thereof, dogwood extracts and the like. The Corni fructus extract comprises methanol extract, ethanol extract, methanol-water mixed solvent extract, and ethanol-water mixed solvent extract.

The existing research shows that the dogwood can improve the T cell subset imbalance in the radiation-induced spleen cells and enhance the immune reconstruction advantage of the Th1 cell subset in the spleen cells after radiation injury, the existing research only inspects the influence on the proportion of the spleen immune cells, and the comprehensive and deep research is not carried out; furthermore, the protective effect of ionizing radiation on other physiological systems, such as hematopoietic system injury, bone marrow system injury and intestinal system injury, is not involved. See: immune reconstitution characteristics of T cell subsets after radiation injury and regulation of traditional Chinese medicine dogwood, the journal of immunology 2015; 000(006): 769-73.

At present, no research report on the effect of dogwood in preparing medicaments for preventing/treating injuries such as ionizing radiation induced hematopoietic injury, bone marrow system injury, intestinal injury, DNA injury and the like is found.

Disclosure of Invention

Aiming at the current situation that medicines for preventing and treating ionizing radiation injury are very limited, the application discloses the application of a dogwood extract in preparing medicines for treating/preventing radiation injury, in particular to the application in treating ionizing radiation injury.

The ionizing radiation includes natural ionizing radiation and artificial radiation. The radiation types include, but are not limited to, X-ray radiation, gamma-ray radiation, heavy ion radiation, alpha-ray radiation, gamma-ray radiation, gamma-ray radiation, gamma-radiation, ⁶⁰ co ray, ¹³⁶ Cs、 ¹⁹² Ir, etc. The specific scenarios include: the medicine for preventing and treating radiation damage to body during radiotherapy and chemotherapy of tumor and nuclear material leakage.

Meanwhile, the application discloses application of the dogwood in preparation of medicines for preventing/treating injuries caused by ionizing radiation, particularly hematopoietic system injuries, bone marrow injuries, radioactive intestinal injuries and the like.

The invention provides a novel radiation injury treatment/protection medicament, which can relieve the current situation that radiation prevention/treatment medicaments are very scarce.

Drawings

FIG. 1 is a diagram showing the radical scavenging ability of the extract of Cornus officinalis.

FIG. 2 is a graph showing the effect of Corni fructus extract on the viability, apoptosis and reactive oxygen species level of bone marrow cells: A. bone marrow cell viability; bone marrow apoptosis level; C. bone marrow cell reactive oxygen levels.

FIG. 3 is a graph showing the effect of extract of Cornus officinalis on the activity of IEC-6 cells as epithelial cells in the irradiated intestinal tract.

FIG. 4 graph of the effect of Corni fructus extract on peripheral blood and bone marrow cell counts in the control mice: A. peripheral blood count results of mice; B. bone marrow cell count results in mice.

FIG. 5 graph of the effect of Cornus officinalis extract on reactive oxygen species levels and DNA damage in BRNC, HPC and HSC of control mice: A. reactive oxygen species levels in mouse BRNC, HPC and HSC; B. γ H2AX expression in mouse BMNC, HPC and HSC.

Detailed Description

The invention is further described with reference to the following figures and examples:

and (3) extracting dogwood fruit: offered by Tianjin medical university;

illumination source: ¹³⁷ a Cs gamma irradiation source provided by the institute of radiology, academy of Chinese medical sciences;

experimental animals: c57BL/6 mouse, purchased from Beijing Huafukang, Inc.;

reagent: 1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH) was purchased from Chinesia; CellTiter-Lumi ^TM The luminescence method cell viability detection kit and the active oxygen detection kit are purchased from Shanghai Biyuntian biotechnology limited company; annexin V-FITC/PI apoptosis detection kit was purchased from Shanghai assist in bioscience;

the instrument comprises: flow cytometry, full-automatic blood analyzer, microplate reader, cell counter, centrifuge, super clean bench, etc.

Example 1 preparation of extract of Cornus officinalis

100g of dogwood is extracted twice by refluxing with 800ml of methanol for 1.5 hours each time. Mixing extractive solutions, and drying under reduced pressure to obtain Corni fructus methanol extract.

EXAMPLE 2 free radical scavenging Activity assay

The antioxidant capacity of the extract of Cornus officinalis was measured by DPPH free radical scavenging method. Dissolving Corni fructus extract in anhydrous ethanol at concentrations of 1, 2, 3, 4, and 5mg/ml respectively. The blank samples were filled with the same volume of innumerable ethanol. After incubation for 30 minutes in the absence of light, the absorbance of the sample at 517nm was measured. The results are shown in figure 1 of the specification.

The results show that the extract of Cornus officinalis has a strong scavenging ability for free radicals in the range of 1 to 5mg/ml, and that the scavenging ability is improved with increasing concentration in the range of 1 to 3 mg/ml.

Example 3 Effect on irradiated bone marrow cell viability, apoptosis and ROS levels

Extracting mouse bone marrow cells, culturing in vitro, adding Corni fructus extract, incubating for 30min, irradiating at an irradiation dose of 1.0Gy, culturing for 18h, and passing through CellTiter-Lumi ^TM Detecting the cell activity by a luminescence method; the bone marrow cells are given 0.001mg/ml of Corni fructus extract 1 hr before irradiation,after 18 hours of culture, bone marrow cells were stained for apoptosis and ROS levels, and the results are shown in FIG. 2 (FIG. 2: A. bone marrow cell viability; B. bone marrow cell apoptosis level; C. bone marrow cell reactive oxygen species level).

The results show that 0.001mg/ml and 0.01mg/ml of the dogwood extract have no toxic effect on bone marrow cells, the cell viability of the bone marrow cells is remarkably reduced after irradiation (p is less than 0.01), and the cell viability of the irradiated bone marrow cells can be remarkably improved by administering 0.001mg/ml of the dogwood extract (p is less than 0.01). Compared with a control group, the irradiation causes the apoptosis of the bone marrow cells and the ROS level to be increased, the average P is less than 0.01, the dogwood extract can obviously reduce the apoptosis of the irradiated bone marrow cells and the ROS level, the average P is less than 0.01, and the dogwood extract has obvious radiation protection effect.

Example 4 Effect on IEC-6 cell viability of irradiated intestinal epithelial cells

Culturing IEC-6 intestinal epithelial cells, adding the dogwood extract, incubating for 30min, irradiating at 9.0Gy dose, culturing for 18h, and detecting cell viability through cck-8; the results are shown in figure 3IEC-6 cell viability.

The results show that 0.001, 0.005 and 0.01mg/ml of the dogwood extract has no toxic effect on IEC-6 cells, the cell viability of IE-6 cells is obviously reduced after irradiation (p is less than 0.01), and the cell viability of the irradiated IEC-6 cells can be obviously improved by administering 0.005 and 0.01mg/ml of the dogwood extract (p is less than 0.01), which indicates that the dogwood extract has obvious radiation protection effect on intestinal tract cells.

Example 5 Effect on peripheral blood and bone marrow cell count in radiation-exposed mice

C57BL/6J male mice were randomly divided into a control group, an irradiated group, and a dogwood + irradiated group, with 5 mice per group. The irradiation dose was 2.0Gy, and control mice were sham irradiated (0 Gy). The Corni fructus extract is administered by gavage 1h before irradiation at a dose of 400 mg/kg, and the control group and the irradiation group are administered with equal amount of normal saline. After 7 days of continuous administration after irradiation, peripheral blood and bone marrow cells of the mice were counted, and the results are shown in the attached figure 4 (FIG. 4: A. peripheral blood count results of mice; B. bone marrow cell count results of mice).

The results show that the irradiation causes the number of White Blood Cells (WBC), Red Blood Cells (RBC) and Platelets (PLT) in the peripheral blood of the mice to be remarkably reduced (P < 0.01), and the number of unilateral femoral nucleated cells (BMNC), Hematopoietic Progenitor Cells (HPC) and Hematopoietic Stem Cells (HSC) is also reduced to different degrees; the extract of Cornus officinalis improves the number of HPC and HSC in peripheral blood cells and bone marrow, wherein the increase of the number of RBC and HPC is statistically significant (P < 0.05). The result shows that the dogwood extract can promote the recovery of peripheral blood and bone marrow cells of mice after radiation exposure and has certain treatment effect on hematopoietic system injury caused by radiation.

Example 6 Effect on reactive oxygen species levels and DNA Damage in radiation-exposed mice BMNC, HPC and HSC

The groups of mice, the dose of irradiation and the administration of the dogwood extract are shown in example 4. After the irradiation was completed, mouse bone marrow cells were taken and examined for ROS levels and DNA damage, and the results are shown in FIG. 5 (FIG. 5: A. reactive oxygen species levels in mouse BMNC, HPC and HSC; B. gamma H2AX expression in mouse BMNC, HPC and HSC).

The results show that the irradiation increases the reactive oxygen species level of HPC and HSC in mice, and the reactive oxygen species level of the cornel + irradiated group is decreased compared to the irradiated group, but there is no statistical difference. DNA damage caused by radiation to BMNC, HPC and HSC was examined by analysis of gamma H2AX expression, and it was found that irradiation significantly increased gamma H2AX expression (P < 0.01) in HPC and HSC and that Cornus officinalis extract significantly decreased gamma H2AX expression (P < 0.05) in HPC and HSC. The above results suggest that the protective effect of the extract of Cornus officinalis on the radiation damage of hematopoietic system caused by ionizing radiation may be achieved by reducing the DNA damage in HPC and HSC.

Claims (6)

1. Application of Corni fructus in preparing medicine for preventing/treating radioactive hematopoietic system injury caused by ionizing radiation is provided.

2. Application of Corni fructus in preparing medicine for preventing/treating radioactive bone marrow injury caused by ionizing radiation is provided.

3. Application of Corni fructus in preparing medicine for preventing/treating radioactive intestinal injury caused by ionizing radiation is provided.

4. Application of Corni fructus in preparing medicine for preventing/treating body DNA injury caused by ionizing radiation is provided.

5. Use according to claims 1-4, characterized by the use in the preparation of an adjuvant drug for tumor radiotherapy.

6. Use according to claim 5, characterized in that the tumors include, but are not limited to, intestinal tumors, hematopoietic tumors, myelomas.

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