CN114533710A - Application of curcumin in preparation of medicine for preventing and treating reproductive system injury caused by ionizing radiation - Google Patents
Application of curcumin in preparation of medicine for preventing and treating reproductive system injury caused by ionizing radiation Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
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Abstract
The invention discloses application of curcumin or pharmaceutically acceptable salts thereof in preparing a medicament for preventing and/or treating reproductive system injury caused by ionizing radiation. The invention adopts cell and animal experiments to verify the performance of curcumin, and the experimental results show that: curcumin has no toxicity to mouse spermatocyte within the concentration range of 1-16 umol/L; 4-12umol/L curcumin can remarkably inhibit the reduction of spermatocyte viability caused by ionizing radiation; curcumin of 10 mu mol/L can obviously improve the apoptosis of spermatocyte caused by ionizing radiation; the testis injury of BALB/c mice caused by 4Gy irradiation of cobalt 60 rays can be reduced by 80mg/kg of preventive administration, which shows that curcumin has the medicinal activity of protecting reproductive system injury caused by ionizing radiation, and can be used for preparing a medicament for inhibiting the reproductive system injury.
Description
Technical Field
The invention relates to the technical field of medicines related to reproductive system injury, in particular to application of curcumin in preparation of medicines for preventing and/or treating reproductive system injury caused by ionizing radiation.
Background
With the development of science and technology, the utilization of atomic energy is increasing. With the widespread use of atomic energy in agriculture, medicine, and military, there is an increasing chance that people will be exposed to ionizing radiation and cause injury. Ionizing radiation can induce a variety of biological damage, and the consequences of ionizing radiation in humans can be serious or even fatal.
Because the spermatids of the testis renew very rapidly, the testis is very sensitive to ionizing radiation and is an important target organ for ionizing radiation. Scientific research shows that ionizing radiation can interfere the normal proliferation and differentiation process of spermatogenic cells, can cause physiological index changes such as sperm malformation, reduction in the number of spermatogenic cells, reduction in the activity and the like, and finally can cause infertility. The number of sperms in the semen of the adult male can be reduced by 0.45Gy of X-ray, and the semen-lacking symptom can be caused by 6Gy of X-ray, and the semen can be recovered after more than two years of X-ray. But the recovered spermatogenic cells may also be associated with DNA damage, resulting in a reduced chance of conception. Even if the next generation is born by an assisted reproductive technique, the offspring are at higher risk of developing the genetic disease than the normal person. Therefore, the development of the protective medicine for the reproductive system injury caused by ionizing radiation is very important for the construction of a nuclear safety guarantee system.
The radioprotectant is a compound which is given a certain substance in an early stage before and after the body is irradiated by ionizing radiation to reduce the radiation damage and promote the recovery of the body, and the compound with the effect is called the radioprotectant. Some existing compounds for effectively preventing injury mainly aim at complications occurring after radiation injury, are not preventive medicaments in the true sense, are mostly sulfur-containing substances, have high toxicity and limit the exertion of the effects of the sulfur-containing substances. Therefore, it is expected that products with low toxicity and being used for clinical prevention and treatment can be developed.
The traditional Chinese medicine has the characteristics of wide source, small toxic and side effect, low price, easy acquisition and the like, and has wide prospect of finding the traditional Chinese medicine which can play a role in resisting radiation. Curcumin is a naturally occurring phytochemical compound of the plant curcuma longa, has been used for thousands of years in many traditional drug therapies, has multiple effects of benefiting gallbladder, reducing blood fat, resisting tumors, resisting oxidation, regulating immunity and the like, but has not been reported as a preventive drug for relieving radiation injury.
Disclosure of Invention
In order to overcome the problems in the prior art, based on the fact that ionizing radiation directly or indirectly acts on cells, the structure and stability of DNA in the cells can be changed, and a large amount of ROS can be generated, so that a plurality of apoptosis signal molecules in the cells can be excited, and apoptosis can be caused, and curcumin has an anti-oxidation effect, and can prevent damage of excessive oxidants generated by ionizing radiation irradiation to a reproductive system. The invention provides a new application of curcumin in ionizing radiation protection, which can be used for preparing a medicament for preventing or treating the injury of ionizing radiation to a reproductive system.
In order to achieve the purpose, the invention adopts the following technical scheme:
the first aspect of the invention provides the application of curcumin or pharmaceutically acceptable salt thereof in preparing a medicament for preventing and/or treating reproductive system injury caused by ionizing radiation.
Further, the curcumin or a pharmaceutically acceptable salt thereof has a pharmaceutical activity of protecting against ionizing radiation damage.
Further, the ionizing radiation includes cobalt 60 radiation, and specifically may be radiation of an intensity of 0.45Gy to 6Gy, more specifically 4 Gy.
Further, the reproductive system injury comprises: decreased spermatocyte activity, increased spermatocyte apoptosis, and damaged testis.
Further, the drug is a drug for inhibiting the decline of the activity of spermatocytes caused by ionizing radiation; and/or, the drug is a drug that ameliorates spermatocyte apoptosis caused by ionizing radiation; and/or the drug is a drug for reducing testicular damage caused by radiation irradiation.
Further, the pharmaceutically acceptable salt is a salt formed by curcumin and an acid of an inorganic acid, an organic acid or an acidic amino acid.
The second aspect of the invention provides a medicament for preventing and/or treating reproductive system injury caused by ionizing radiation, wherein the medicament is a preparation prepared by taking curcumin or pharmaceutically acceptable salts thereof as an active ingredient, and the preparation further comprises pharmaceutically acceptable medicinal auxiliary ingredients and/or pharmaceutical excipients.
Further, the preparation comprises 1-99 wt% of curcumin or pharmaceutically acceptable salts thereof and 99-1 wt% of medicinal auxiliary components and/or medicinal excipients, preferably 10-80 wt% of curcumin or pharmaceutically acceptable salts thereof and 90-20 wt% of medicinal auxiliary components and/or medicinal excipients. It is understood that the above-mentioned pharmaceutically acceptable auxiliary ingredients and/or pharmaceutical excipients are all agents conventionally used in the art, such as dimethyl sulfoxide, sodium carboxymethyl cellulose aqueous solution, tween, cyclodextrin and the like.
Further, the types of the preparation include liquid preparation, granule, capsule, tablet, granule, sustained release agent or injection.
Further, in certain embodiments, the formulation is a liquid formulation made by dissolving curcumin in dimethyl sulfoxide, or made by dissolving curcumin in an aqueous solution of sodium carboxymethyl cellulose. More specifically, curcumin was dissolved in dimethyl sulfoxide, and the concentration of curcumin was 20 mmol/L; curcumin is dissolved in sodium carboxymethylcellulose water solution, and the concentration of curcumin is 80 mg/mL.
Further, the pharmaceutically acceptable salt is a salt formed by curcumin and an acid of an inorganic acid, an organic acid or an acidic amino acid.
Further, the inorganic acid includes hydrochloric acid, phosphoric acid, nitric acid, etc., the organic acid includes formic acid, acetic acid, maleic acid, citric acid, methanesulfonic acid, etc., and the acidic amino acid is aspartic acid or glutamic acid.
The third aspect of the invention provides application of a plant taking curcumin as a main component in preparing a medicament or food for preventing and/or treating reproductive system injury caused by ionizing radiation.
Compared with the prior art, the invention has the following beneficial effects by adopting the technical scheme:
the invention discovers the new medical application of the curcumin for the first time, and the curcumin can be used for preventing and treating the reproductive system injury caused by ionizing radiation;
the curcumin has wide source and small toxic and side effects, and experimental verification shows that the 16 mu mol/L drug concentration is used for treating mouse spermatocyte, the cell state is good, no obvious adverse reaction exists when the curcumin is treated under the weight concentration of 80mg/kg, and repeated verification is carried out on the activity of the curcumin for resisting the reproductive system damage caused by ionizing radiation through the cell level and the animal level, and the experimental result shows that the curcumin can inhibit the reduction of the activity of the mouse spermatocyte caused by cobalt 60 irradiation, improve the apoptosis of the spermatocyte caused by the cobalt 60 irradiation and reduce the BALB/c mouse testicle damage caused by the cobalt 60 irradiation.
The curcumin adopted by the invention can be a beneficial supplement of a high-efficiency low-toxicity anti-ionizing radiation induced reproductive system injury medicine, and has a good development prospect.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention without limiting the invention. In the drawings:
FIG. 1 is a graph showing the toxicity of curcumin to spermatocytes in one embodiment of the present invention;
FIG. 2 is a graph showing the results of the protective effect of different concentrations of curcumin on the reduction in cell viability caused by ionizing radiation in one embodiment of the present invention;
FIG. 3 is a graph showing the results of the protective effect of curcumin on the process of apoptosis by ionizing radiation in one embodiment of the present invention;
fig. 4 is a graph showing the protective effect of curcumin on reduction of sperm concentration in mice caused by ionizing radiation in one embodiment of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. The experimental methods of the following examples, which are not specified under specific conditions, are generally determined according to national standards. The experimental materials not shown in the following examples are all commercially available materials. The equipment used in each step in the following examples is conventional. If there is no corresponding national standard, it is carried out according to the usual international standards, to the conventional conditions or to the conditions recommended by the manufacturer. Unless otherwise indicated, all parts are parts by weight and all percentages are percentages by mass. Unless defined or indicated otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention.
It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict. The invention is further described with reference to the following drawings and specific examples, which are not intended to be limiting.
In the following embodiments, the radioactive source used is 60Co gamma ray source of navy military medical university, and the dosage rate is 0.8 Gy/min; the irradiation method for experimental animals comprises placing mice in a plastic box, and subjecting to one-time whole body irradiation of 4Gy 60Co gamma; in cell experiments, curcumin (curcumin) is dissolved in dimethyl sulfoxide, and the concentration of mother liquor is 20 mmol/L.
Example 1 cytotoxicity assay of curcumin on mouse spermatocytes
In the embodiment, mouse spermatocyte GC-2 which is generally recognized for the genital system injury in vitro research is selected as a model. Dissolving curcumin in dimethyl sulfoxide, wherein the concentration of mother liquor is 20mmol/L, preparing curcumin solutions with the use concentrations of 4, 8, 12, 16 and 20 mu mol/L by adopting a stepwise dilution method, and the dilution solution is DMEM culture solution for culturing cells. The control group was treated with an equal volume of dimethyl sulfoxide to a 20. mu. mol/L syringaldehyde solution. GC-2 cells are inoculated in a 96-well plate, the curcumin solution and the dimethyl sulfoxide with different concentrations are respectively added when the cells are in a logarithmic growth phase, and the growth condition of the cells is detected by CCK8(Dojindo, operation method follows instructions) after 48 hours of treatment.
The detection result of the experiment is shown in figure 1, the curcumin concentration below 16 mu mol/L still does not obviously change the proliferation rate of the GC-2 cells, namely the curcumin concentration does not influence the activity of the GC-2 cells.
Example 2 protective Effect of curcumin on reduction of mouse spermatocyte viability by ionizing radiation
In this example, GC-2 cells (1X 10 cells) were collected5one/mL) was inoculated into a 96-well plate at 100. mu.L/well of drug (curcumin solution) at a final concentration of 12, 8, 4. mu. mol/L in 5 wells of each concentration, and a blank control cell suspension and an irradiation cell suspension were added. 4Gy irradiation is carried out after 2 hours of drug action, and the cell growth condition is detected by adopting CCK8 after 24 hours of irradiation.
The detection results of the above experiments are shown in fig. 2, the spermatocyte viability of curcumin irradiated by gamma ray 4Gy at 4-12 μmol/L is increased by more than 10%, which shows that curcumin pretreatment plays a protective role in spermatocyte viability reduction caused by ionizing radiation.
Example 3 protective Effect of curcumin on apoptosis of mouse spermatocytes caused by ionizing radiation
In this example, GC-2 cells were seeded in 6-well culture plates, and four groups were set up: (1) no treatment group; (2) curcumin group at 10 μmol/L; (3)4Gy ionizing radiation group; (4)4Gy ionizing radiation + 10. mu. mol/L curcumin group. 24h after treating the cells according to the experimental conditions, the cell culture solution and the cells after trypsinization are collected by a centrifuge tube, centrifuged at 2000 Xg for 5 minutes, the supernatant is discarded, and the cells are resuspended in PBS and counted. 5-10 ten thousand of the resuspended cells were centrifuged at 1000 Xg for 5 minutes, the supernatant was discarded, and 195. mu. Lannexin V-FITC conjugate was added to gently resuspend the cells. Add 5. mu.L of Annexin V-FITC and mix gently. Add 10. mu.L of propidium iodide staining solution and mix gently. Incubate for 10 minutes at room temperature in the dark and detect with flow cytometry.
The detection results of the above experiments are shown in fig. 3, the apoptosis of the cells is not affected by pure curcumin treatment, and the apoptosis rate of the GC-2 cells can be remarkably reduced by adopting 10 mu mol/L curcumin treatment.
Example 4 protective Effect of curcumin on sperm concentration reduction in BALB/c mice caused by ionizing radiation
This example dissolved curcumin in a 1% by mass aqueous solution of sodium carboxymethylcellulose, wherein the concentration of curcumin was 80 mg/mL.
Eight-week-old male BALB/c mice were acclimatized and divided into 4 groups as needed: normal group, dosing group (curcumin), radiation group and radiation + dosing group (curcumin), 5 per group. The blank group and the radiation group mice are given sodium carboxymethylcellulose aqueous solution with the same volume and the mass concentration of 1 percent. Curcumin solution was administered by gavage at a dose of 80mg/kg for 8 weeks. After 9 days of 4Gy ionizing radiation, mice are killed by cervical dislocation, and testicles and epididymis of the mice on both sides are separated, and tunica mesenterium and fat around the testicles and epididymis are removed. Cutting epididymis head into several segments, incubating in water bath at 37 deg.C for 10min to allow sperm to disperse automatically. The pipette gun pipetted 10. mu.L of liquid, dropped on a hemocytometer and counted.
The detection results of the above experiments are shown in fig. 4, the pure curcumin treatment does not affect the sperm concentration of the mouse, and the curcumin treatment can significantly improve the sperm concentration of the mouse.
As can be seen from the above examples, curcumin has no toxicity to mouse spermatocyte in the concentration range of 1-16 umol/L; 4-12umol/L curcumin can remarkably inhibit the reduction of spermatocyte viability caused by ionizing radiation; curcumin of 10 mu mol/L can obviously improve the apoptosis of spermatocyte caused by ionizing radiation; the testis injury of BALB/c mice caused by 4Gy irradiation of cobalt 60 rays can be reduced by 80mg/kg of preventive administration, and the result shows that curcumin has the medicinal activity of protecting reproductive system injury caused by ionizing radiation, and can be used for preparing the medicines for inhibiting the reproductive system injury.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.
Claims (10)
1. Application of curcumin or pharmaceutically acceptable salts thereof in preparing medicines for preventing and/or treating reproductive system injury caused by ionizing radiation.
2. The use as claimed in claim 1, wherein the curcumin or pharmaceutically acceptable salt thereof has pharmaceutical activity against damage to the reproductive system caused by ionizing radiation.
3. Use according to claim 1, wherein the ionising radiation comprises cobalt 60 radiation.
4. The use of claim 1, wherein the medicament is a medicament for inhibiting a decline in spermatocyte viability caused by ionizing radiation; and/or, the drug is a drug that ameliorates spermatocyte apoptosis caused by ionizing radiation; and/or the drug is a drug for reducing testicular damage caused by radiation irradiation.
5. The use according to any one of claims 1 to 4, wherein the pharmaceutically acceptable salt is a salt of curcumin with an acid of an inorganic acid, an organic acid or an acidic amino acid.
6. The medicine for preventing and/or treating reproductive system injury caused by ionizing radiation is a preparation prepared by taking curcumin or pharmaceutically acceptable salts thereof as an active ingredient, and further comprises pharmaceutically acceptable medicinal auxiliary ingredients and/or medicinal excipients.
7. The medicament for preventing and/or treating the injury of the reproductive system caused by ionizing radiation according to claim 6, wherein the preparation comprises 1-99 wt% of curcumin or a pharmaceutically acceptable salt thereof and 99-1 wt% of a pharmaceutically acceptable auxiliary ingredient and/or a pharmaceutical excipient.
8. The medicament for preventing and/or treating genital system injury caused by ionizing radiation according to claim 6, wherein the type of the preparation comprises liquid preparation, granule, capsule, tablet, granule, sustained-release preparation or injection.
9. The medicament for preventing and/or treating reproductive system injury caused by ionizing radiation according to any one of claims 6 to 8, wherein the pharmaceutically acceptable salt is a salt of curcumin and an acid of an inorganic acid, an organic acid or an acidic amino acid.
10. Application of plant containing curcumin as main ingredient in preparing medicine or food for preventing and/or treating reproductive system injury caused by ionizing radiation is provided.
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