CN111529557A - Application of plant fruit oil in preparation of medicines and health foods for preventing ovarian injury caused by radiotherapy and chemotherapy - Google Patents
Application of plant fruit oil in preparation of medicines and health foods for preventing ovarian injury caused by radiotherapy and chemotherapy Download PDFInfo
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- CN111529557A CN111529557A CN202010525333.1A CN202010525333A CN111529557A CN 111529557 A CN111529557 A CN 111529557A CN 202010525333 A CN202010525333 A CN 202010525333A CN 111529557 A CN111529557 A CN 111529557A
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/08—Drugs for genital or sexual disorders; Contraceptives for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
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- A—HUMAN NECESSITIES
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Abstract
The application discloses application of plant fruit oil in preparation of medicines and health-care foods for preventing ovarian damage caused by radiotherapy and chemotherapy, wherein the plant fruit oil is sea buckthorn seed oil. The application finds that the sea buckthorn seed oil has a good prevention effect on ovarian injury caused by radiotherapy and chemotherapy. The sea buckthorn seed oil can prevent the ovarian injury and the disturbance of the estrus cycle of mice caused by radiotherapy and chemotherapy, and can effectively improve the ovarian reserve function and the like. The invention provides a basis for directly utilizing the seabuckthorn seed oil to prevent ovarian injury caused by radiotherapy and chemotherapy.
Description
Technical Field
The application relates to the technical field of application of plant fruit oil, in particular to application of plant fruit oil in preparing medicines and health-care foods for preventing ovarian damage caused by radiotherapy and chemotherapy.
Background
Although generally, the tumor incidence in men is higher than in women, the tumor incidence in women increases very rapidly with the current change of lifestyle. Recently, there has been a report: in the next two decades, the incidence of cancer in uk women is 6 times higher than in men, and this phenomenon is not only in the west but also in our countries, so that the terms breast cancer, cervical cancer, ovarian cancer, gastric cancer, etc. are no longer unknown to us, and the burden of the neoplastic disease is very heavy both for the individual patient and for the whole society. However, for tumors, the treatments that we can select are often very limited. At present, patients who can be treated by surgery mostly adopt surgical resection assistance to carry out radiotherapy and chemotherapy, and the radiotherapy and chemotherapy become the biggest choice for patients who lose surgical chances. Although the radiotherapy and chemotherapy improve the symptoms of patients to a certain extent and prolong life, the damage of radiotherapy and chemotherapy to other organs of the body is always an inevitable problem in clinic. The ovary is the most sensitive part to chemotherapy and radiation in the female reproductive system, and the ovary can be damaged by large-dose and long-time radiotherapy and chemotherapy, so that the ovary is damaged. In recent years, due to the rapid increase of the number of tumors suffered by women, ovarian functions of patients are damaged when the patients receive chemoradiotherapy, perimenopause symptoms such as menstrual disorder and amenorrhea occur, even infertility can be caused to bring great influence on physical and mental health of the women, but for the influence, the restraint is often not done clinically because of the lack of effective preventive medicines. The world health organization estimates that two-fifths of the 1400 ten thousand cancers diagnosed per year are preventable. Therefore, it is important to take a series of measures to protect the structure and function of the ovary before applying a treatment for damage to the ovary. At present, the effect of ovarian protection measures is not ideal, and better medicines are required to be searched for to deal with the damage, so that how to protect the ovarian function of patients undergoing radiotherapy and chemotherapy from being damaged is paid more and more attention by researchers.
Currently, the most studied protective drugs include gonadotropin releasing analogues, oral contraceptives, SIP, amifostine and other antioxidants. However, gonadotropin releasing analogues such as triptorelin and oral contraceptives belong to hormone drugs, and symptoms such as hot flashes, night sweats and the like caused by osteoporosis and low estrogen states can appear after long-term administration; exogenous antioxidant drugs are difficult to be popularized clinically due to factors such as high price, many side effects of drugs, uncertain long-term treatment effect and the like. Therefore, drug developers and clinicians try to find a comprehensive, effective, cheap and convenient drug which can reduce and prevent the damage of ovaries caused by radiotherapy and chemotherapy.
The Chinese medicine seabuckthorn has a long medicinal history in China, and the seabuckthorn has the functions of relieving cough and asthma, promoting blood circulation and removing blood stasis and the like which are recorded in the four medical dictionaries. More than 400 active ingredients of seabuckthorn fruit mainly comprise: nutrient and active ingredients such as flavonoids, crude fibers, polysaccharides, amino acids, vitamins, trace elements, carotene, proteins, organic acids, chlorophyll and the like; can be widely applied to various fields of national economy such as food, medicine, light industry, spaceflight, agriculture, animal husbandry, fishery and the like. The sea-buckthorn seed oil is the main component of sea-buckthorn fruit, is rich in nutrition, contains more than 100 active substances such as various vitamins, fatty acids, trace elements, phytosterol, linolein, sea-buckthorn flavone, alpha-tocopherol, superoxide and the like and various amino acids required by a human body according to determination, has the effects of resisting inflammation, promoting granulation, repairing ulcer, inhibiting tumor and resisting cancer, and particularly has obvious curative effects on cerebral arteriosclerosis, angina, hyperlipidemia and the like. So far, no prevention effect of sea buckthorn seed oil on ovarian injury caused by radiotherapy and chemotherapy is reported.
Disclosure of Invention
The application discloses application of plant fruit oil in preparing a medicament and health-care food for preventing ovarian damage caused by radiotherapy and chemotherapy, namely application of sea buckthorn seed oil in preparing a medicament and health-care food for preventing ovarian damage caused by radiotherapy and chemotherapy.
According to the embodiment of the application, the application of the plant fruit oil in preparing the medicine for preventing the ovarian damage caused by the radiotherapy and chemotherapy is provided, and the plant fruit oil is the seabuckthorn seed oil. .
According to the embodiment of the application, the application of the plant fruit oil in preparing the health-care food for preventing the ovarian damage caused by the radiotherapy and chemotherapy is provided, and the plant fruit oil is the seabuckthorn seed oil.
Furthermore, the dosage form of the medicine comprises decoction, soft capsules, dripping pills, emulsion, tablets, lotion, liniment, ointment or suppository.
Furthermore, the dosage form of the health food comprises decoction, soft capsules, dripping pills, emulsion, tablets, lotion, liniment, ointment or suppository.
The application proves that the seabuckthorn seed oil has new functions completely different from the functions described above for the first time: the sea buckthorn seed oil has good prevention effect on ovarian injury caused by radiotherapy and chemotherapy. The sea buckthorn seed oil can prevent the ovarian injury and the disturbance of the estrus cycle of mice caused by radiotherapy and chemotherapy, and can effectively improve the ovarian reserve function and the like. The application provides a basis for directly utilizing the seabuckthorn seed oil to prevent ovarian injury caused by radiotherapy and chemotherapy.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 is an experimental design of the present application;
FIG. 2 is a technical roadmap for the present application;
FIG. 3 shows the H & E staining results of paraffin sections of the ovarian tissues of various groups of mice;
FIG. 4 is the vaginal secretion observation results of the mice in each group;
FIG. 5 is a graph of the change in estrogen E2 levels in groups of mice;
FIG. 6 shows the results of immunohistochemical AMH expression in ovarian tissues of various groups of mice;
FIG. 7 shows the change in serum AMH content of each group of mice.
Detailed Description
The application provides an application of plant fruit oil in preparing a medicine for preventing ovarian injury caused by radiotherapy and chemotherapy, wherein the plant fruit oil is sea buckthorn seed oil.
The application provides an application of plant fruit oil in preparing health food for preventing ovarian damage caused by radiotherapy and chemotherapy, wherein the plant fruit oil is sea buckthorn seed oil.
Furthermore, the dosage form of the medicine comprises decoction, soft capsules, dripping pills, emulsion, tablets, lotion, liniment, ointment or suppository.
Furthermore, the dosage form of the health food comprises decoction, soft capsules, dripping pills, emulsion, tablets, lotion, liniment, ointment or suppository.
The application proves that the seabuckthorn seed oil has new functions completely different from the functions described above for the first time: the sea buckthorn seed oil has good prevention effect on ovarian injury caused by radiotherapy and chemotherapy. The sea buckthorn seed oil can prevent the ovarian injury and the disturbance of the estrus cycle of mice caused by radiotherapy and chemotherapy, and can effectively improve the ovarian reserve function and the like. The application provides a basis for directly utilizing the seabuckthorn seed oil to prevent ovarian injury caused by radiotherapy and chemotherapy.
The application aims to discuss the protective effect of the preventive application of the seabuckthorn seed oil on ovarian tissue damage caused by radiotherapy and chemotherapy, and provides an effective protection method and scientific basis for ovarian damage caused by application of a large amount of chemotherapy drugs in clinical tumor treatment.
In order to confirm that the seabuckthorn seed oil has the new functions, the following experiments are carried out.
1. Technical scheme
(1) Establishing an animal model of ovarian injury caused by chemoradiotherapy drugs:
the experimental design is shown in figure 1.
Cyclophosphamide (CY) and Busulfan (BUS) are common anticancer drugs in clinic and are also representative ovarian toxicity drugs. In the experiment, an ovary injury mouse model is established by injecting 120mg/kg of cyclophosphamide and 10mg/kg of busulfan into the abdominal cavity of an adult female mouse at one time, and the continuously intragastric administration of 2.5ml/kg of seabuckthorn seed oil is carried out on the ovary injury mouse every day from 7 days before the modeling to 18 days after the modeling to serve as a seabuckthorn seed oil prevention group (seabuckthorn seed oil protection group); meanwhile, GnRHA triptorelin (38mg/kg) is injected subcutaneously for one time 7 days before modeling to serve as a positive control group (GnRHA group), normal saline (0.2mL/d) is used for replacing CY + BUS to be continuously perfused with stomach to serve as an ovary injury control group (model group) (the perfusion in each day is consistent with that of the model group and a sea buckthorn seed oil prevention group, and the difference caused by the perfusion operation is avoided), and a normal control group is additionally arranged, namely, a normal mouse without ovary injury is perfused with normal saline with the same volume. Materials are taken 18 days after molding, the histomorphology change of the ovary is observed, the number of all levels of follicles in the ovary tissue is counted, the level of serum E2 and anti-mullerian hormone (AMH) is detected, and the expression level of AMH protein is detected by immunohistochemistry.
(2) Intervention in an ovarian injury animal model induced by chemotherapeutic agents:
the experiment is divided into a normal control group, a model group, a positive control group (GnRh-a (triptorelin)), and a seabuckthorn seed oil prevention group. After the success of molding is confirmed, the normal control group and the model group are intragastrically administered with the same amount of saline water every day, and the seabuckthorn seed oil group is intragastrically administered with 2.5ml/kg (0.1ml) of seabuckthorn seed oil every day for 18 days.
(3) Experimental results monitoring and data collection
Mice in each group were tested every 4 days and recorded for weight, feeding and estrous cycle changes. On day 18 after the treatment, the material was taken and the ovarian morphological changes were observed by paraffin embedding medium section staining technique. Changes in AMH were detected by immunohistochemical techniques.
2. Technical route map
A technical roadmap is shown in fig. 2.
3. Main techniques
1) Animal model for establishing ovarian injury caused by chemotherapeutic drugs
Cyclophosphamide (CY) and Busulfan (BUS) are common anticancer drugs in clinic and are also representative ovarian toxicity drugs. In the research, an ovary injury mouse model is established by injecting 120mg/kg cyclophosphamide and 10mg/kg busulfan into the abdominal cavity of an adult female mouse at one time.
2) Examination of estrus cycle by smear method of vaginal cast-off cell flushing fluid
Sucking a proper amount of normal saline by a dropper, washing and sucking vaginal secretion on a glass slide at the same time point every 4 days, rapidly observing cell morphology under a microscope, and judging estrus cycle.
4. Results of the experiment
4.1 the seabuckthorn seed oil can prevent the lesion of the mouse ovarian tissue caused by radiotherapy and chemotherapy.
Chemotherapy can directly result in damage to ovarian tissue. Previous studies have demonstrated that chemotherapy causes ovarian volume reduction, cortical atrophy, follicular loss, interstitial fibrosis and hyalinization, granulosa cells manifest acute necrotic lesions, such as a reduction in the number of granulosa cells, the appearance of numerous myelin-like structures, intracellular and intercellular edema, severe arteriosclerosis of the ovaries, marked endomysial hyperplasia and often total luminal occlusion.
As shown in fig. 3, (1) general observations: the ovary of the normal control group mouse is normal in shape and ruddy in color; the ovary of the model group mouse is atrophic, the surface is pale, and single or more cystic follicles can be seen on the surface of the ovary; the ovary surface of the seabuckthorn seed oil mouse is smooth and ruddy and is close to the normal tissue. (2) And (4) observing under a mirror: the ovary of a normal control group mouse can see a plurality of follicles and corpus luteum at different development stages under HE staining, granular cells are complete in shape and are regularly arranged, and the number of the granular cells is 8-9; the ovary of the model group mouse can see a plurality of primary follicles and a small number of secondary follicles, no egg cells are seen in the follicles close to the mature period, the number of visible granular cell layers is thinned, the arrangement is loose, part of the follicle falls off, no radial crown is seen, the number of visible granular cell layers can be reduced to 5-6, the corpus luteum is less, the ovarian atresia is increased and is saccular expansion, the follicular membrane cells are proliferated, the oocyte in the follicular is disappeared, and the layer of the follicular membrane cells is thicker; the ovarian tissue follicle of the positive control group is intact and is close to the normal tissue, but the interstitial substance still has obvious fibrosis; the ovary of the seabuckthorn seed oil mouse can be provided with a corpus luteum and a plurality of follicles at different development stages, the follicles close to the maturation stage can be provided with ova and a radial crown, the arrangement of granular cells is compact, the number of layers is increased, the cell layer of the follicular membrane is thin, and the shape of the ovarian follicle is close to that of a normal control group.
4.2 the seabuckthorn seed oil can prevent the disturbance of the estrus cycle of mice caused by radiotherapy and chemotherapy.
The estrus cycle refers to the time interval from the start of the last estrus to the start of the next estrus, or the last ovulation to the next ovulation, in a female mammal. The oestrus cycle is generally divided into a prophase oestrus (promestrus), an oestrus, a postoestrus (postoestrus) and an interestrus (interval) according to several main aspects including the mental state of the animal, the sexual desire performance of the female on the male, the growth and development of follicles in the ovaries, the ovulation and the formation of corpus luteum, and the physiological changes of the female reproductive tract (uterus, vagina, etc.). Major accessory organs such as ovary, uterus and vagina are constantly changing during the oestrus cycle. Having a regular estrus cycle is a marker of sexual maturity in mammals, meaning that the ovaries begin to function. In patients with ovarian impairment, menstrual disorders are one of the most typical clinical manifestations, manifested as infrequent menstruation, scanty menstruation and even amenorrhea. The expression in mice is indicative of a disturbance in the estrous cycle, i.e., a sustained interval.
As shown in fig. 4, the prematurity (positive control) was observed with spindle-shaped epithelial cells in clusters or alone, and with fewer leukocytes. In estrus (normal control group), a large amount of squamous epithelial cells are seen, the shape is large and irregular, no nucleus is formed, and white blood cells disappear; in the anaphase of estrus (seabuckthorn seed oil), various cell types, white blood cells, anucleate epithelial cells and fusiform epithelial cells can be seen; large numbers of leukocytes and very few fusiform epithelial cells were visible during estrus (model group). After the molding, the observation of the continuous estrus cycle every 4 days shows that: the control group of mice all have obvious 4-5 d estrus cycles and are distributed relatively evenly in each phase; the model group mice do not show obvious existence of estrus cycle, more continuous estrus intervals can be seen, and other phases of the estrus cycle show less expression, thus prompting no ovulation of the estrus cycle disorder; the sea-buckthorn seed oil group mice have obvious estrus cycles, and the recovery of the estrus cycles after the sea-buckthorn seed oil is applied is prompted. Therefore, the sea-buckthorn seed oil has a good prevention effect on the ovarian injury mouse estrus cycle disorder caused by radiotherapy and chemotherapy.
4.3 the sea-buckthorn seed oil can prevent the decline of the level of the estrogen E2 of mice caused by radiotherapy and chemotherapy.
The exact mechanism by which chemotherapy drugs cause ovarian dysfunction is not yet fully understood, and it is now mainly believed that the primary route by which chemotherapy causes ovarian dysfunction may be the promotion of follicular cell apoptosis. It is generally believed that actively dividing cells are more sensitive to the action of chemotherapeutic drugs which may cause abnormal growth or disappearance of follicles, affecting secretion of relevant hormones, and other symptoms secondary to hormonal dyssecretion, such as decreased estrogen secretion which may feedback to hypothalamic GnRH. Increased secretion of pituitary gonadotropins (FSH, LH), which in turn accelerates the development of primary follicles into growing follicles, so that the growing follicles are destroyed again by the chemotherapeutic drugs, forming a vicious circle, eventually leading to the end of ovarian failure with thirst from the primary follicle reserve.
As shown in fig. 5: the levels of estrogen E2 in the serum of mice in the normal control group, the sea buckthorn seed oil group and the positive control group are similar and are all obviously higher than those in the model group, and the sea buckthorn seed oil group is slightly higher than that in the positive control group, which further proves that the sea buckthorn seed oil is a better medicament for preventing ovarian injury caused by radiotherapy and chemotherapy and has definite curative effect.
4.4 the seabuckthorn seed oil can prevent the hypofunction of the ovary reserve of mice caused by radiotherapy and chemotherapy.
AMH is secreted by granulosa cells of pre-antral follicles and antral follicles, inhibits the growth of the follicles, prevents the premature consumption of the follicles, and preserves the reserve of ovaries, so that the AMH is the best method for detecting the reserve function of the ovaries under different clinical conditions at present. When follicular granulosa cells are abundant and ovarian function is better, the level of AMH will be at a higher level. Conversely, as follicular granulosa cells gradually decrease, ovarian function gradually deteriorates and the level of AMH also gradually decreases. AMH is used as the best index for evaluating the ovarian reserve function, and the higher AMH index indicates that the larger the stock of the ova, the stronger the fertility is relatively. Decreased AMH suggests that the ovaries are aging, indicating a decline in female fertility.
Immunohistochemistry was used to detect AMH expression in ovarian tissue. As shown in fig. 6, there was slight yellow staining around the follicles and between granulosa cells in the normal control group, demonstrating AMH expression; the model group had little visible yellow-stained parts; in the positive control group and the seabuckthorn seed oil group, large areas of yellow stain are formed around the follicles and among granular cells, which indicates that the AMH is highly expressed in the ovaries of the two groups of mice. It can be seen that the yellow-staining fraction in the seabuckthorn oil group was nearly equal to the positive control group, which tells us that seabuckthorn oil had nearly the same efficacy as triptorelin in ovarian reserve.
4.5 the sea buckthorn seed oil can prevent the reduction of the AMH hormone content of mice caused by radiotherapy and chemotherapy.
The present application uses an enzyme-linked immunosorbent assay (ELISA) to determine serum AMH levels. As shown in figure 7, compared with the model group, the seabuckthorn seed oil can obviously improve the expression of AMH in blood plasma of mice with ovarian injury caused by chemotherapy, and further proves that the seabuckthorn seed oil has a good protective effect on ovarian reserve function reduction caused by chemotherapeutic drugs.
As can be seen from the above experimental results,
(1) the oleum Hippophae can prevent change of ovary tissue structure of mouse caused by radiotherapy and chemotherapy, and improve ovary tissue morphology
(2) The oleum Hippophae can prevent the disturbance of estrus cycle of mice caused by radiotherapy and chemotherapy, and maintain normal physiological cycle of mice
(3) The oleum Hippophae can be used for preventing hypoestrogenic level of mice caused by radiotherapy and chemotherapy
(4) The oleum Hippophae can prevent the damage of ovary reserve function of mice caused by radiotherapy and chemotherapy
(5) The similarity of radiotherapy and chemotherapy to the ovarian injury of mice, and the seabuckthorn seed oil has the prevention and protection effects on the ovarian dysfunction caused by radiotherapy and chemotherapy.
Other embodiments of the present application will be apparent to those skilled in the art from consideration of the specification and practice of the application disclosed herein. This application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the application and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the application being indicated by the following claims.
It is to be understood that the present application is not limited to what has been described above, and that various modifications and changes may be made without departing from the scope thereof. The scope of the application is limited only by the appended claims.
Claims (4)
1. An application of a plant fruit oil in preparing a medicament for preventing ovarian injury caused by radiotherapy and chemotherapy is disclosed, wherein the plant fruit oil is sea buckthorn seed oil.
2. An application of a plant fruit oil in preparing a health food for preventing ovarian injury caused by radiotherapy and chemotherapy is disclosed, wherein the plant fruit oil is sea buckthorn seed oil.
3. The use of claim 1, wherein the medicament is in the form of decoction, soft capsule, dripping pill, emulsion, tablet, lotion, liniment, ointment or suppository.
4. The use of claim 2, wherein the health food is in the form of decoction, soft capsule, dripping pill, emulsion, tablet, lotion, liniment, ointment or suppository.
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JP2007023012A (en) * | 2005-07-20 | 2007-02-01 | Reino:Kk | Skin cream containing hippophae oil and enzymes |
CN110075139A (en) * | 2019-05-29 | 2019-08-02 | 宁夏医科大学 | Application of the Seabuckthorm Seed Oil in prevention Stein-Leventhal syndrome drug |
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