Efficacy, safety and recurrence of new progestins and selective progesterone receptor modulator for the treatment of endometriosis: a comparison study in mice

Abstract

Background: Current medical treatments for endometriosis are very limited. Progestin and selective progesterone receptor modulators (SPRM) are developed but their efficacy, safety, mechanism and recurrence in endometriosis are not fully studied.

Methods: In order to compare therapeutic, side effects and therapeutic actions of Esmya, Duphaston and Dienogest in endometriosis. Experimental endometriosis was induced by either intraperitoneal or subcutaneous mouse endometrium transplantation. Lesion size, weight and histology at the end of intervention were compared. Expression of related markers in the endometriotic lesions were examined. Body, uterus and ovary weights, endometrial glands and thickness (ETI), and follicle count were measured. For recurrent study, lesion growth before and after intervention was monitored.

Results: After Esmya, Duphaston, Dienogest treatment, lesion size and weight were significantly decreased. Proliferation Pcna expression was significantly decreased in all groups, but proliferation cells were significantly decreased only in Duphaston group. Apoptosis Mapk1 expression and TUNEL-positive cells were significantly increased in Duphaston group. Adhesion Mmp2 and Itgavβ3 expression were significantly increased in Esmya group. Plau, Hif1α and Vegfa expression, peritoneal fluid PGE2 levels, and ERα and ERβ expression were not affected; while PR expression was significantly lower in all groups. Endometrial gland count in uterus was significantly increased in Dienogest group, ETI was significantly decreased in Duphaston group, and AFC were significantly increased in Esmya group. Upon treatment cessation, lesion growth rebound quickly in Dienogest and Duphaston groups, but slowly in Esmya group.

Conclusion: Esmya, Duphaston and Dienogest are effective anti-endometriosis drugs targeting proliferation, apoptosis and adhesion. Esmya, Duphaston and Dienogest are all well tolerable, although endometrial glandular hyperplasia was found in Dienogest, endometrial atrophy in Duphaston, follicle accumulation in Esmya.

Keywords: Endometrium; Progestin; Selective progesterone receptor modulator.

Fig. 1

Esmya, Duphaston and Dienogest suppress the growth and development of endometriotic lesions in mice. a Lesion size was measured by caliper at the end of intervention. Representative in situ pictures of the endometriotic lesions in vehicle, Esmya, Duphaston and Dienogest group are shown. Red arrows indicate the locations of the lesions. b Endometriotic lesion size and weight. Data are shown as mean ± SEM, n = 9 for each group, *: p < 0.05 when compared to the control group. c Histological change of lesions after medication. This panels show the representative cross-sections of the endometriotic lesion as endometrial cyst-like structure. Scar bar: 400um. d Proliferation of the endometriotic lesions. Representative pictures of immunohistochemical staining of Ki67 in endometriotic-like lesions in different groups after treatment. Scar bar: 200um. e Comparison of Ki67 positively stained epithelial and stroma cells in the lesions. f Representative pictures of TUNEL staining in endometriotic-like lesions in different groups after treatment. Scar bar: 200um. g Comparison of TUNEL positively cells in the lesions. Data are shown as mean ± SEM, n = 3 for each group, *: p < 0.05 when compared to the control group

Fig. 2

Comparisons of anti-endometriosis mechanism of Esmya, Duphaston and Dienogest. Relative mRNA expression of a-c, adhesion (Mmp2 and Itgav β3) and invasion (Plau) marker genes, d-f, apoptosis-related genes (Mapk1, NF-kB p105 and pro-Caspase 3), g-i, angiogenesis and proliferation mrakers (Vegf, Hif1α and Pcna) and j-k, hormone receptors (ERα/ERβ ratio and PR) in lesions; and ELISA quantitation of l, Prostaglandin E2 (PGE2) concentrations in peritoneal fluid. Data are shown as mean ± SEM, n = 9 for each group, *: p < 0.05 when compared to the control group

Fig. 3

Safety profiles of Esmya, Duphaston and Dienogest. a Body weight changes during the study intervention. Data was recorded every 5 days from day 1 to day 25 after transplantation. b Serum estrogen and progesterone concentrations after treatment in the mice. c Upper panels show the representative cross section images of uterus, structure of endometrium gland, stroma and lumen cavity. Scar bar: 200um. Lower panels compare the uterus size, ratio of uterus and body weight, endometrium thickness index (ETI) and uterus gland count from left to right. d Upper panels show the histological change of ovary after treatment. Scar bar: 200um. Lower panels compared the ovary size, ratio of ovary and body weight and the follicle count from left to right. Data are shown as mean ± SEM. *: p < 0.05 when compared to the control group

Fig. 4

Dynamic change of endometriotic lesion size during and after treatment of Esmya, Duphaston and Dienogest. The endometriotic lesions were transplanted subcutaneously in day − 7. Intervention was started at day 0 and withdrawn at day 21 (yellow dot line). Data were shown as mean ± SEM, n = 10 for each group, *: p < 0.05 when compared to the control group in the corresponding days and groups