CN107982269B

CN107982269B - Application of crocetin in preparation of medicine for preventing and treating polycystic ovarian syndrome

Info

Publication number: CN107982269B
Application number: CN201711146515.2A
Authority: CN
Inventors: 盛亮; 金晶; 胡巧云
Original assignee: Nanjing Medical University
Current assignee: Nanjing Medical University
Priority date: 2017-11-17
Filing date: 2017-11-17
Publication date: 2020-03-17
Anticipated expiration: 2037-11-17
Also published as: CN107982269A

Abstract

The invention discloses application of crocetin in preparation of a medicine for preventing and treating polycystic ovarian syndrome. On the basis of the existing efficacy of crocetin, a polycystic ovary syndrome mouse model induced by dihydrotestosterone in a fetal period is adopted to deeply research the treatment effect of the crocetin. The results show that the crocetin can obviously improve the estrus cycle rhythm disorder of a polycystic ovarian syndrome model mouse, reduce the weight of ovaries and the number of locked ovaries, increase the number of corpus luteum, large antral follicles and preovulatory follicles, obviously improve the levels of estradiol, luteinizing hormone, follicle stimulating hormone, progesterone and testosterone of the polycystic ovarian syndrome model mouse, and has good effect of treating the polycystic ovarian syndrome. In addition, in the experimental process, no obvious adverse reaction occurs, and the safety is high.

Description

Application of crocetin in preparation of medicine for preventing and treating polycystic ovarian syndrome

Technical Field

The invention relates to a new application of a traditional Chinese medicine monomer crocetin in treating polycystic ovarian syndrome, belonging to the technical field of medicines.

Background

Polycystic ovarian syndrome is a common disorder of the reproductive endocrine system affecting 4-8% of women in the reproductive age. It is mainly characterized in that: thin hair, ovulatory or anovulation, hyperandrogenic clinical manifestations (hirsutism, acne, etc.) and/or hyperandrogenism, ovarian polycystic changes. Anovulatory hemorrhage or menstruation at adolescence, ovulation disorder at birth is an important reason for infertility, and risks of diabetes, non-alcoholic fatty liver, hyperlipidemia, hypertension, gestational diabetes, gestational hypertension and endometrial cancer can be increased at a long term. Is a disease with multiple phenotypes, difficult cure and double risks of near term and long term.

Polycystic ovarian syndrome is currently treated primarily with hormone modulating drugs. Dandelion 35 is suitable for patients with high androgen clinical manifestations or high androgen blood diseases, and is the most common medicine for lowering androgen at present. Periodic withdrawal of progestogen and other short-acting oral contraceptives may also be employed to regulate the menstrual cycle. Non-hormonal treatments have also entered the line of sight in recent years due to the many contraindications and side effects of hormonal treatment. For example, vitamin D levels are often low in patients with polycystic ovarian syndrome clinically, and vitamin D supplementation is considered as a method for ameliorating polycystic ovarian syndrome.

The traditional Chinese medicine considers that the polycystic ovarian syndrome belongs to the ranges of infertility, hypomenorrhea, delayed menstrual period, amenorrhea, abdominal mass and the like. The primary cause of this disease is the disorder of zang-fu organs, with turbid phlegm and blood stasis as the secondary causes. The disease is closely related to kidney, liver and spleen, but mainly caused by kidney deficiency and spleen deficiency, and pathogenic products such as phlegm dampness and blood stasis act on the body to cause the disorder of the genital axis of kidney-Tian Dec-Chong and ren-uterus. The existing Chinese medicine treatment prescription and syndrome are combined into one addition and subtraction.

Saffron is a perennial flower of crocus of the family iridaceae, and is also a common spice. Has tranquilizing, expectorant, and spasmolytic effects, and can be used for treating gastropathy, menstruation regulating, measles, fever, jaundice, and hepatosplenomegaly. Crocetin is one of the main components of stigma croci Sativi, and can also be extracted from fructus Gardeniae. It is reported that crocetin can be used for treating tumors such as breast cancer, lung cancer, pancreatic cancer and the like, and can also be used for treating Alzheimer disease, insulin resistance and cardiovascular diseases. However, no report related to the application of crocetin in treating polycystic ovarian syndrome exists at present.

Disclosure of Invention

The purpose of the invention is as follows: on the basis of the existing application of crocetin, the invention develops new clinical application of crocetin through a large number of experimental screens, and provides new application of crocetin in preventing and treating polycystic ovarian syndrome.

The technical scheme is as follows: the invention provides application of crocetin in preparing a medicine for preventing and treating polycystic ovarian syndrome.

The structural formula of crocetin is:

molecular formula C₂₀H₂₄O₄。

The invention applies dihydrotestosterone to pregnant mice to induce offspring female mouse polycystic ovary syndrome. After the progeny have matured and developed polycystic ovarian syndrome, they are treated with crocetin. H & E staining is used for observing the number of atretic ovary, luteal phase, antral follicle and mature antral follicle, the weight of the ovary, the estrus cycle and the levels of estradiol, luteinizing hormone, follicle stimulating hormone, progesterone and testosterone, and the result is used as an evaluation index of the polycystic ovary syndrome.

The experimental result shows that the crocetin can obviously improve the estrus cycle rhythm disorder of a polycystic ovarian syndrome model mouse, reduce the weight of ovaries and the number of locked ovaries, increase the number of corpus luteum, antral follicles and mature antral follicles, obviously improve the levels of estradiol, luteinizing hormone, follicle stimulating hormone, progesterone and testosterone of the polycystic ovarian syndrome model mouse, and has good effect of preventing and treating polycystic ovarian syndrome. The crocetin can obviously improve the estrus cycle rhythm disorder of a polycystic ovarian syndrome model mouse, reduce the weight of ovaries and the number of locked ovaries, increase the number of corpus luteum, antral follicles and mature antral follicles, obviously improve the levels of estradiol, luteinizing hormone, follicle stimulating hormone, progesterone and testosterone of the polycystic ovarian syndrome model mouse, and has good effect of preventing and treating polycystic ovarian syndrome.

Drawings

Figure 1 is a bar graph of the change in body weight and ovarian weight of mice following testosterone making and crocetin treatment.

FIG. 2 is a graph of the effect of crocetin on the estrous cycle of polycystic ovary syndrome mice.

FIG. 3 is a bar graph of the effect of crocetin on the estrous cycle of polycystic ovary syndrome mice.

FIG. 4 is a bar graph of the effect of crocetin on the number of atretic follicles, corpus luteum, large sinus follicles, and preovulatory follicles in a polycystic ovary syndrome mouse.

Figure 5 is a bar graph of levels of luteinizing hormone, follitropin, estradiol, testosterone and progesterone in mouse serum after dihydrotestosterone modelling and crocetin treatment.

Detailed Description

The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the specific material ratios, process conditions and results thereof described in the examples are illustrative only and should not be taken as limiting the invention as detailed in the claims.

Example 1 therapeutic effect of crocetin on dihydrotestosterone induced polycystic ovary syndrome in mice.

1. Experimental Material

1.1 drugs and reagents for experiments

Crocetin is available from shanxi forest friend natural products, ltd.

1.2 Experimental animals and raising

Adult male and female mice (C57Bl/6J × A/J) F1(B6A) were used in this study, provided by Kavens, and certification numbers: 201602427. all mice are raised in an SPF animal house with 12-hour light and shade alternation, 22-24 ℃ and 55-65% of humidity. Free water intake.

2. Method of producing a composite material

2.1 Dihydrotestosterone-induced polycystic ovary syndrome mouse model modeling method

The male and female mice were housed and injected daily with dihydrotestosterone (250 μ g) subcutaneously during 16-18 days of gestation, while the control mice were injected with the solvent.

2.2 therapeutic effect of dihydrotestosterone crocetin on polycystic ovary syndrome mice.

Offspring females grew up to 8 weeks, treatment groups: the administration was continued for 5 weeks with a dose of 50mg/kg/d of gastric crocetin. Mice were sacrificed at either pre-estrus or inter-estrus. The model group and the control group were both given physiological saline.

2.3 mouse POCS index

[1] Mouse ovary and body weight: the mice were weighed before sacrifice, and ovaries were harvested after sacrifice and weighed.

[2] And (3) determining the estrus cycle of the mouse: after the female mice mature sexually, the reproductive organs are proliferated and degenerated periodically and changed into an estrus period (4-5 d). Generally divided into four phases, namely an estrus early phase (0.5-1.5 d), an estrus phase (1-3 d), an estrus late phase (1-3 d) and an estrus interval (2-4 d). The characteristics of mouse vaginal cast-off cells in different estrus cycles are different, and the estrus cycle of the mouse is judged by vaginal cell smear observation:

the prophase of estrus is all nucleated epithelial cells and occasionally non-nucleated keratinized epithelial cells

The estrus is composed of anucleated keratinized epithelial cells with a small number of nucleated epithelial cells in between

Leucocyte and keratinized epithelial cell decrease appear in the anaphase of estrus, and all three cells have

Large amount of leucocytes, small amount of nucleated epithelial cells and mucous membrane in estrus

[3] And (3) observing the structure of the ovary of the mouse: one side of the ovaries was fixed by formalin immersion for 24H, paraffin embedded, H & E stained, followed by pathological structural analysis.

[4] And (3) serum hormone detection: the serum kit detects the levels of estradiol, luteinizing hormone, follicle stimulating hormone, progesterone and testosterone in the serum of the mouse at the prophase of estrus.

2.4 statistical analysis

Data mining

Expressed, tested using one-way analysis of variance (ANOVA), p<A difference of 0.05 is statistically significant.

3. Results of the experiment

3.1 crocetin reduces ovarian weight gain in polycystic ovarian syndrome mice.

The experimental results in fig. 1 show that the weights of the mice in the control group, the model group and the administration group are not significantly different, but the weight of the ovary in the model group is significantly increased (p <0.05), and the results show that the crocetin can significantly reverse the abnormal weight gain of the ovary (p < 0.05).

3.2 crocetin improves the estrus cycle of polycystic ovary syndrome mice.

The experimental results of fig. 2 and 3 show that the estrous cycle of the model group mice is prolonged remarkably in total and is expressed by the increase of the postestrous and estrous interval (p < 0.05). After crocetin treatment, the anagen, the interestrous and the total estrous cycle were all significantly shortened (# p < 0.05).

3.3 crocetin improves pathological structures of polycystic ovary syndrome mouse ovary

From the experimental results in figure 4, it was shown that the modelling of dihydrotestosterone caused a significant increase in the number of atretic follicles in mice, while reducing the number of corpus luteum, antrum follicles and preovulatory follicles (. p < 0.05). After the treatment with crocetin, the number of atretic follicles decreased significantly, and the number of corpus luteum, antrum magnum follicles and preovulatory follicles increased significantly (# p < 0.05).

3.4 crocetin improves polycystic ovary syndrome mouse hormone levels

The experimental results in fig. 5 show that the serum of the model group mice has obviously increased luteinizing hormone and testosterone, and the serum of the model group mice has obviously decreased estradiol, follicle stimulating hormone and progesterone (p < 0.05). Treatment with crocetin reverses the abnormal elevation of luteinizing hormone and testosterone levels and elevates lower estradiol, follitropin and progesterone levels (# p < 0.05).

The invention adopts dihydrotestosterone to treat pregnant mice pregnant for 16-18 days, so as to cause female offspring polycystic ovary syndrome. The ovary weight of a model mouse is increased, the estrus cycle is disordered, luteinizing hormone and testosterone are increased, and estradiol, follicle stimulating hormone and progesterone are obviously reduced; a significant increase in the number of atretic follicles, a decrease in the number of corpus luteum, large antral follicles and preovulatory follicles. The model can well simulate typical symptoms of polycystic ovarian syndrome, and has good reproducibility, effectiveness and usability. The pharmacodynamic test results show that the crocetin obviously reduces abnormal ovarian weight gain, improves the estrus cycle and disorder of serum hormone level, finally reduces the number of atretic follicles, and up-regulates the number of corpus luteum, antrum magnum follicles and preovulatory follicles, and shows good efficacy of treating polycystic ovarian syndrome.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims

1. Application of crocetin in preparing medicine for preventing and treating polycystic ovarian syndrome is provided.

2. The use of crocetin for the preparation of medicaments for the prevention and treatment of polycystic ovary syndrome according to claim 1, wherein crocetin and pharmaceutically acceptable carriers are prepared into tablets, granules, pills, capsules, oral liquids or injections.