阅读说明：本技术 一种治疗早期子宫内膜异位症的中药单体及其应用 (Traditional Chinese medicine monomer for treating early endometriosis and application thereof ) 是由 曹阳 张婷婷 朱焰 庄梦斐 崔金刚 张瑜 杜尘 王馨悦 王唯迪 于 2020-08-10 设计创作，主要内容包括：本发明涉及一种治疗早期子宫内膜异位症的中药单体及其应用,所述中药单体是人参皂苷Rg3。结果表明,人参皂苷Rg3能抑制子宫内膜异位症大鼠的异位内膜的血管生成,但又不会影响在位内膜的形态,说明生理性的血管未受其影响,故其对正常组织毒性极小。因此,人参皂苷Rg3可应用于子宫内膜异位症的早期治疗及腹腔镜术后消除残留术中残留的病灶的治疗,防止复发又不妨碍排卵,符合理想的早期治疗及预防内异症发生的药物。另人参皂苷Rg3能明显下调内异症大鼠血清E2的水平,但对血清P水平无显著影响,提示其可能通过调节体内激素水平,调整体内内分泌环境,在治疗疾病的同时不影响受孕,同时怀孕本身也是对EMs最有效的治疗,实用性强。(The invention relates to a traditional Chinese medicine monomer for treating early endometriosis and application thereof, wherein the traditional Chinese medicine monomer is ginsenoside Rg 3. The results show that the ginsenoside Rg3 can inhibit the angiogenesis of the ectopic intima of the rat with endometriosis, but can not affect the shape of the ectopic intima, which indicates that physiological blood vessels are not affected by the angiogenesis, so the toxicity to normal tissues is extremely low. Therefore, the ginsenoside Rg3 can be applied to the early treatment of endometriosis and the treatment of eliminating residual focus in operation after laparoscopy, prevents relapse without obstructing ovulation, and meets the ideal medicine for early treatment and prevention of endometriosis. In addition, the ginsenoside Rg3 can obviously reduce the level of the serum E2 of rats with the endometriosis, but has no obvious influence on the level of serum P, which suggests that the ginsenoside Rg3 can regulate the hormone level in vivo and adjust the endocrine environment in vivo by regulating the hormone level in vivo, does not influence conception while treating diseases, is the most effective treatment on EMs during pregnancy, and has strong practicability.)

1. Application of Chinese medicinal monomer ginsenoside Rg3 in preparing medicine for treating early stage endometriosis is provided.

2. Application of traditional Chinese medicine monomer ginsenoside Rg3 in preparation of medicine for inhibiting ectopic intima volume and angiogenesis of rat with endometriosis.

3. Application of traditional Chinese medicine monomer ginsenoside Rg3 in preparation of medicine for eliminating residual focus in laparoscopy.

4. Application of traditional Chinese medicine monomer ginsenoside Rg3 in preparation of medicine for reducing level of serum E2 of early-stage endometriosis patients is provided.

5. Application of traditional Chinese medicine monomer ginsenoside Rg3 in preparing medicine for regulating hormone level in early-stage endometriosis patients is provided.

6. Application of traditional Chinese medicine monomer ginsenoside Rg3 in preparation of medicine for promoting ovulation of early-stage endometriosis patients is provided.

7. Application of traditional Chinese medicine monomer ginsenoside Rg3 in preparation of medicine for improving conception of early-stage endometriosis patients is provided.

8. The use of any one of claims 1 to 7, wherein the medicament comprises monomeric ginsenoside Rg3 and pharmaceutically acceptable adjuvants.

9. A medicine for treating early endometriosis is characterized in that the medicine is prepared from traditional Chinese medicine monomer ginsenoside Rg3 and pharmaceutically acceptable auxiliary agents.

10. The pharmaceutical composition of claim 9, wherein the pharmaceutical composition comprises granules, powders, capsules, tablets, mixtures or oral liquids.

Technical Field

The invention relates to the technical field of application of traditional Chinese medicine monomers, in particular to a traditional Chinese medicine monomer for treating early endometriosis and application thereof.

Background

Endometriosis (EMs) is a persistent disease characterized by pelvic pain and infertility caused by the presence of endometrium with growth function outside the uterine cavity, common and difficult gynecological diseases. The most typical symptoms of the disease are secondary and progressive lower abdominal pain and lumbosacral pain, which usually occur 1-2 days before menstruation, are the most severe in 1 st day after menstruation, and are accompanied by feeling of anal tenesmus and distension, and usually disappear in the clear menstruation. The etiology and pathogenesis of endometriosis are complex and changeable, and complications such as pain, infertility and ovarian cancer induction risk seriously affect the life quality of women. The pathogenesis of endometriosis is not clear clinically, researchers think that the endometriosis is caused by menstrual blood countercurrent implantation, and research data show that the pathogenesis of endometriosis is related to factors such as blood vessel-machine lymphatic metastasis, coelomic epithelium biochemical treatment and the like. The occurrence and development of the disease are reported to involve a plurality of pathological processes such as autophagic function deficiency, ectopic cell anoikis, angiogenesis, inflammation, steroid hormone generation, oxidative stress and the like, which also determine the complexity and difficulty of the disease, and particularly have more obstacles to the diagnosis agent treatment of early diseases.

Research shows that a process is needed for development of the EMs from dysmenorrhea or pelvic pain, the EMs in early stage do not necessarily combine typical clinical symptoms, patients often see a doctor only when the dysmenorrhea is aggravated or pelvic masses are combined, the time of about 7 years is needed for forming EMs focuses seen by naked eyes in human bodies, how to fully discriminate the EMs in early stage of occurrence and perform effective intervention to cut off the pathological development process of the EMs in time, and the method has important significance for improving the effective rate, preventing the EMs and relieving the pain of the patients.

Traditional Chinese medicine has unique advantages in this respect. The research shows that the ginsenoside Rg3 can inhibit the generation of the blood vessels of the ectopic intima of the rat with endometriosis, but can not affect the shape of the in-situ intima, so that the physiological blood vessels are not affected by the influence of the ginsenoside Rg3, and the toxicity to normal tissues is very low. Therefore, the ginsenoside Rg3 can be applied to the early treatment of endometriosis and the treatment of eliminating residual focus in operation after laparoscopy, prevents relapse without obstructing ovulation, and meets the ideal medicine for early treatment and prevention of endometriosis. In addition, ginsenoside Rg3 can obviously reduce the level of E2 in the serum of rats with the endometriosis, but has no obvious influence on the level of P in the serum, which suggests that the ginsenoside Rg3 can regulate the hormone level in vivo and adjust the endocrine environment in vivo by regulating the hormone level in vivo, does not influence conception while treating diseases, and simultaneously, the pregnancy is the most effective treatment on EMs.

In summary, the subject group finds that the ginsenoside Rg3 can be used for early treatment of endometriosis, preventing the occurrence of endometriosis, and promoting conception during administration, and is an effective traditional Chinese medicine for treating endometriosis. Other studies on the treatment of internal disorders with this drug have not been reported.

Disclosure of Invention

The invention aims to provide application of a traditional Chinese medicine monomer ginsenoside Rg3 aiming at the defects of the prior art.

It is still another object of the present invention to provide a medicament for treating early endometriosis.

In order to achieve the purpose, the invention adopts the technical scheme that:

in a first aspect, the invention provides application of a traditional Chinese medicine monomer ginsenoside Rg3 in preparation of a medicine for treating early endometriosis.

In a second aspect, the invention provides application of a traditional Chinese medicine monomer ginsenoside Rg3 in preparation of a medicine for inhibiting the volume and angiogenesis of an ectopic intima of a rat with endometriosis.

In a third aspect, the invention provides application of a traditional Chinese medicine monomer ginsenoside Rg3 in preparation of a medicine for eliminating residual focus in a surgery after a laparoscopic surgery.

In a fourth aspect, the invention provides application of a traditional Chinese medicine monomer ginsenoside Rg3 in preparation of a medicine for reducing the level of serum E2 of a patient with early endometriosis.

In a fifth aspect, the invention provides application of a traditional Chinese medicine monomer ginsenoside Rg3 in preparing a medicine for regulating hormone level in an early-stage endometriosis patient.

In a sixth aspect, the invention provides application of a traditional Chinese medicine monomer ginsenoside Rg3 in preparation of a medicine for promoting ovulation of patients with early endometriosis.

In a seventh aspect, the invention provides application of a traditional Chinese medicine monomer ginsenoside Rg3 in preparation of a medicine for improving pregnancy of patients with early endometriosis.

Preferably, the medicine comprises traditional Chinese medicine monomer ginsenoside Rg3 and pharmaceutically acceptable auxiliary agents.

In order to achieve the second object, the invention adopts the technical scheme that:

a medicine for treating early endometriosis is prepared from ginsenoside Rg3 as monomer and pharmacologically acceptable auxiliaries.

Preferably, the pharmaceutical preparation comprises granules, powders, capsules, tablets, mixtures or oral liquids.

The ginsenoside Rg3 has the following molecular formula:

the invention has the advantages that:

the invention firstly provides the application of the ginsenoside Rg3 in treating early endometriosis. As is well known to those skilled in the art, endometriosis is a common and difficult gynecological disease, persistent pathological changes characterized by pelvic pain and infertility are caused by the appearance of endometrium with growth function outside uterine cavity, research shows that development of dysmenorrhea or pelvic pain into EMs requires a process, EMs at early stage do not necessarily have typical clinical symptoms, patients often see a diagnosis only because of aggravation of dysmenorrhea or pelvic mass, and EMs focus seen by naked eyes in human bodies needs about 7 years, because the etiology and mechanism of the disease are complex, many in the prior art, late diagnosis and treatment of endometriosis are related, but treatment of early endometriosis is very little, the invention provides a new traditional Chinese medicine monomer for early treatment of endometriosis, and experimental results show that: ginsenoside Rg3 can inhibit the volume and angiogenesis of endometrium of rat with endometriosis, but will not affect the shape of endometrium, so physiological blood vessel is not affected by it, so it has little toxicity to normal tissue. Therefore, the ginsenoside Rg3 can be applied to the early treatment of endometriosis and the treatment of eliminating residual focus in operation after laparoscopy, prevents relapse without hindering ovulation, and meets the ideal medicine for treating endometriosis infertility. In addition, ginsenoside Rg3 can obviously reduce the level of E2 in the serum of rats with the endometriosis, but has no obvious influence on the level of P in the serum, which suggests that the ginsenoside Rg3 can regulate the endocrine environment in vivo by regulating the level of hormone in vivo and does not influence conception while treating diseases.

In addition, the research also shows that the ginsenoside Rg3 can inhibit the angiogenesis of endometriosis through inhibiting a PI3K/Akt/mTOR signal channel mediated by VEGFR-2, and the latest research on the channel shows that the static primordial follicle activation of the ovary of an endometriosis patient can be accelerated, namely the decline of the ovary reserve function is related to the abnormal activation of the PI3K/Akt channel, so that the ginsenoside Rg3 also can treat an EMs infertility patient with the decline of the ovary reserve function through inhibiting the channel, provides a powerful basis, relieves the pain of the patient, has strong practicability and good application prospect,

drawings

FIG. 1 is a vaginal smear test of adult female SD rats. A: the estrus vaginal smear is entirely anucleated keratinocytes; b: the estrus vaginal smear has a large amount of white blood cells or a small amount of epithelial cells among the white blood cells and no white blood cells.

FIG. 2 is the morphological observation of the ectopic intima tissue of EMs rats. A: white saccular ectopic intima; b: red nodular ectopic intima; → to ectopic intima.

FIG. 3 is a HE staining assay of endometrial tissue from EMs rats. A: an ectopic intima; b: an in-place inner membrane; → denotes glandular epithelial cells, × 100.

FIG. 4 shows the body weight gain in the individual groups of EMs rats before and after drug treatment. A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (n is 12, P <0.05)

FIG. 5 shows the ectopic intima volumes of EMs rats in the groups before treatment. A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (n is 12)

FIG. 6 shows the ectopic intima volume of EMs rats in each group after treatment. A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (n: 12; P < 0.05; P <0.01)

FIG. 7 shows the inhibition of ectopic intima after each treatment. A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (n is 12, P is 0.01)

FIG. 8 shows the observation of the lesion before and after administration to rats in the model group of EMs. A large amount of liquid in the graft is gathered and is in a transparent sacculus shape, and the sacculus is not obviously reduced after the drug is administrated. A: before treatment, B: and (5) after treatment.

FIG. 9 shows the observation of the lesion before and after treatment in the rats of castrate EMs. A: before castration treatment, B: after castration treatment. The atrophy of the graft disappears after administration.

FIG. 10 shows the lesion observations before and after administration of EMs to rats in the gestrinone (positive control) group. A: before administration of gestrinone, B: after administration of gestrinone. The graft size is obviously reduced after the drug is applied.

FIG. 11 is an observation of the lesion before and after administration of low dose ginsenoside Rg3 group EMs rats. A: prior to administration of the low dose Rg3, B: low dose Rg3 was administered. The graft tends to shrink, but the clear capsule cavity remains clearly visible.

FIG. 12 is an observation of the lesion before and after administration of high dose ginsenoside Rg3 group EMs rats. A: prior to administration of high dose Rg3, B: high dose Rg3 was administered. The graft had substantially collapsed and disappeared after the administration of the drug.

FIG. 13 is a morphological observation (X100) of the in-situ intima of EMs rats, A: low dose of ginsenoside Rg3, B: high dose of ginsenoside Rg3, C: gestrinone group, D: model control group, E: and (4) castration groups.

FIG. 14 shows the change in thickness of the inner membrane at ectopic sites among the groups (. times.100), A: ginsenoside Rg3 low dose group, B: ginsenoside Rg high dose group, C: gestrinone group, D: model control group, E: and (4) castration groups. (→: pointing to the ectopic intima; group B, group C, group E all have significantly lower ectopic intimal epithelial height than group D)

Figure 15 is an illustration of the effect of ginsenoside Rg3 on the height of ectopic intimal gland epithelium, a: low dose group of ginsenoside Rg 3; b: low dose group of ginsenoside Rg 3; c: a group of gestrins; d: a model control group; e: and (4) castration groups. (n ═ 6;. P <0.01 compared to group D).

Figure 16 is a graph of the effect of ginsenoside Rg3 on serum E2 and P levels in EMs rats, a: low dose group of ginsenoside Rg 3; b: low dose group of ginsenoside Rg 3; c: a group of gestrins; d: a model control group; e: and (4) castration groups. (left: E2; right: P; n: 12;. P <0.05 or 0.01 compared to group D).

FIG. 17 is an immunohistochemical assay for VEGF protein in EMs rat ectopic intima tissue, A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (. times 400)

FIG. 18 is an immunohistochemical assay for VEGFR-2 protein in EMs rat ectopic intima tissue. A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (. times 400)

FIG. 19 is an immunohistochemical assay for p-Akt protein in EMs rat ectopic intima tissue. A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (. times 400)

FIG. 20 is an immunohistochemical assay for p-mTOR protein in EMs rat ectopic intima tissue. A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (. times 400)

FIG. 21 shows the average gray value of IHC detection of each protein to be detected in the ectopic intima of EMs rats. A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (p < 0.05;. p < 0.01; compare with group D)

FIG. 22 shows WB detection of VEGF, VEGFR-2, Akt and mTOR proteins in the ectopic intima of EMs rats. A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups.

FIG. 23 shows the relative expression levels of the respective proteins to be tested in the ectopic intima of EMs rats (WB assay). A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (p < 0.05;. p < 0.01; compare with group D)

FIG. 24 shows the relative expression levels of each mRNA tested in the ectopic intima of EMs rats (RT-PCR assay). A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (p < 0.05;. p < 0.01; compare with group D)

FIG. 25 shows the effect of ginsenoside Rg3 on ectopic intima apoptosis. A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (. times 200)

FIG. 26 shows the measurement of apoptotic activity in ectopic intima tissue of EMs rats in each group (TUNEL method). A: low dose Rg3 group; b: high dose Rg3 group; c: a group of gestrins; d: a model group; e: and (4) castration groups. (p <0.05, compare with group D; n ═ 6)

Detailed Description

The invention will be further illustrated with reference to specific embodiments. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Furthermore, it should be understood that various changes and modifications can be made by those skilled in the art after reading the disclosure of the present invention, and equivalents fall within the scope of the appended claims.