RU-2861521-C1 - METHOD FOR MODELLING ENDOMETRIOSIS IN MICE

RU2861521C1RU 2861521 C1RU2861521 C1RU 2861521C1RU-2861521-C1

Abstract

FIELD: experimental medicine; biotechnology. SUBSTANCE: in female CD-1 mice weighing at least 18 g, making an incision in the peritoneum. Next, ligating the uterine horn at both ends, after which it is removed, cut into pieces of the same size, area 4-6 mm 2 . Said pieces are implanted into the peritoneum or mesentery of the same animal. After 14±4 days from the start of the experiment, the animals are sacrificed and the endometriomas obtained at the implantation site are measured. EFFECT: more valid, with short preparation time, model of endometriosis in mice. 3 cl, 2 ex

Inventors

Ilichev Aleksandr Vladimirovich
Belopolskaya Mariya Vladimirovna
Bustios-Guryanova Anna Albetrovna
Areshidze David Aleksandrovich
Lovat Maksim Lvovich
Mikhaleva Lyudmila Mikhajlovna
Kontorshchikov Andrej Sergeevich

Dates

Publication Date: 20260505
Application Date: 20250702

Claims (3)

1. A method for modeling endometriosis in mice, characterized in that surgical modeling of endometriosis is carried out on female mice of the CD-1 line, weighing at least 18 g, whereby an incision is made in the peritoneum, the uterine horn is ligated at both ends, after which the uterine horn is removed, cut into pieces of the same size, with an area of 4-6 mm2 , and the said pieces are implanted into the peritoneum or mesentery of the same animal, after 14±4 days from the start of the experiment, the animals are dissected and the endometriomas obtained at the site of implantation are measured.
2. The method according to paragraph 1, characterized in that the removed uterine horn is cut into 2-3 parts.
3. The method according to paragraph 1, characterized in that the administration of the medicinal product to determine its pharmacological activity begins in the period from the first to the seventh day after the operation.

Description

The invention relates to the field of biotechnology, specifically to modeling endometriosis in laboratory animals. It can be used to study the therapeutic efficacy of drugs or treatments for endometriosis. Endometriosis is a pathological process characterized by the presence of tissue outside the uterine cavity that has morphological and functional properties similar to the endometrium. It is estimated that at least 10% of all women, primarily of reproductive age, suffer from endometriosis. The most significant clinical manifestations of endometriosis are pelvic pain, female infertility, and the presence of endometrioid cysts in the pelvis. Despite extensive study of endometriosis, the etiology of this pathological process remains unclear. Numerous theories have been proposed for the pathogenesis of endometriosis, including retrograde menstruation, metaplastic, embryonic, dyshormonal, and immune imbalance theories. Endometriosis is treated with medication and/or surgery, with the choice of treatment strategy depending on numerous factors (Endometriosis. Clinical Guidelines. // Approved by the Scientific and Practical Council of the Ministry of Health of the Russian Federation. - 2024). The development of new treatments for endometriosis is impossible without experimental animal models that mimic the disease. The spontaneous endometriosis model in primates is considered the best, but the high financial and time costs of working with these animal species limit their use as experimental models. Therefore, disease modeling methods in rodents and rabbits are widely used. Experimental models of endometriosis are based on the transplantation of uterine fragments or endometrial cells into the peritoneal cavity of an animal. Depending on the origin of the grafted tissue, such models are divided into homologous and heterologous (Rodent animal models of endometriosis-associated pain: unmet needs and resources available for improving translational research in endometriosis. / Tejada et al. // International Journal of Molecular Sciences - 2023, 24 , 2422). In the homologous model, endometrial tissue is obtained from the animal uterus and transplanted or dispersed into the abdominal cavity of immunocompetent animals. Such uterine transplantations are performed in rabbits (Increased expression levels of metalloprotease, tissue inhibitor of metalloprotease, metallothionein, and p63 in ectopic endometrium: an animal experimental study. / Brandão et al. // Rev Bras Ginecol Obstet - 2018, 40 , 705-712) and rats (A modern view on the experimental creation of endometriosis in an animal model (own data) / L.V. Adamyan, L.M. Mikhaleva, et al. // Problemy reproductsii 2024, Vol. 30, No. 6, pp. 55-60). The use of transgenic mice with “green fluorescent protein” (GFP) allows for a non-invasive method of identifying and assessing the size and location of endometrioid cysts, and monitoring the angiogenesis of blood vessels surrounding a fragment of the endometrium when it is transplanted into non-fluorescent mice (Transgenic mice applications in the study of endometriosis pathogenesis / Zhao et al. // Front Cell Dev Biol - 2024, 12, 1376414). In a heterologous model, portions of human endometrial biopsies are implanted intraperitoneally or subcutaneously into immunodeficient or immunosuppressed mice. Although the implantation rate is approximately 30%, this percentage can increase to 100% if the implants are sutured or adhered with tissue adhesive. Traditionally, this type of research most often uses athymic mice (Nude-NU), which lack mature T cells. Despite this, the implants cease to function after the fourth week due to B cells appearing in the third week. Solutions to avoid these problems include using other rodent strains, such as SCID (severe combined immunodeficiency) mice or NOD/SCID (severe combined immunodeficiency and diabetes) mice. These mice lack both T cells and functional B cells, have higher implantation rates than thymus-ablated mice, and implants persist for more than 4 weeks. Because of the role of other immune cells, such as NK cells and macrophages, NOD/SCID/γCnull (NOG) or RAG2/CD47/IL2RG mice have been generated that lack B, T, and NK cells and have impaired dendritic and macrophage cell function (Rodent animal models of endometriosis-associated pain: unmet needs and resources available for improving translational research in endometriosis. / Tejada et al. // International Journal of Molecular Sciences - 2023, 24, 2422). A key limitation of these models is the use of immunocompromised mice as recipients, which does not allow a full assessment of immune responses, which play a major role in the etiology of the disease. A method for modeling juvenile endometriosis using a xenograft model is known from the prior art. According to this method, a 2x2 mm fragment of an endometriotic lesion, obtained from women in the early reproductive period, is sutured to the peritoneum of homozygous female Balb/c-nude mice weighin