| Object：Asoprisnil is a selective progesterone receptor modulators, the aim of this part is toexplore the anti-implantation effect of asoprisnil.Methods：Healthy adult female kunming mice, weighing25±5g, were purchased. Vaginal smearwas taken daily, and only estrus female mice were caged individually with fertile males. Positivemating was verified by identifying copulation plugs in vagina, and designated as Day1of pregnancy.Post-coitus mice were randomized divided into vehicle control group, low dosage (0.05mg/10g/d),medium dosage (0.1mg/10g/d) and high dosage (0.2mg/10g/d) asoprisnil group. Post-coitus animalswere gavaged Asoprisnil administered from Day1to Day3of pregnancy, once every day. Animalswere sacrificed on Day8of pregnancy, and then checked the presence or absence of nidation and thenumber of implantation embryos. And their uteri were sampled for hematoxylin and eosin (HE)staining. Implantation rate was calculated using the following formula: Implantation rate= Successful nidation mice/Total test mice×100%, Average number of implantation embryos=Total implantation embryos/Total test mice.Results：①Implantationrateinvehiclecontrolgroup,lowdosegroup,mediumdosegroupand high dose group were respectively as95%,75%,35%and20%. The differences amongmedium dose group, high dose group and vehicle group were statistically significant（P<0.01）. When the implantation rate in medium dose group and high dose group werecompared with the low dose group, the differences were significant (P<0.01).②Theaverage number of embryo implantation site in vehicle control group, low dose group,medium dose group and high dose group were respectively as11.80±3.38,6.40±5.50,4.45±6.44and2.15±4.88. The difference was significant when compared with the vehiclecontrol group, the difference among asoprisnil group were significant (P<0.01).③Invehicle control group, the type of luminal epithelium was monolayer columnar cells, the line wascurved and folded, the gland showed active proliferation; There were plenty of gland and vessels inthe lamina propria, stromal cells were loose scattered and showed decidua change. While inasoprisnil group, the luminal epithelium cells were showed proliferative change, especially inmedium dose group and high dose group, the development of the gland was not significant; Glandsand vessels were rare in the lamina propria, stromal cells were compact distributed withoutsignificant decidua change.Conclusion Asoprisnil can inhibit the embryo implantation in mice by affecting thedecidua change of the stromal cells in endometrium. Objective: Asoprisnil is a member of the selective receptor modulator, the purpose of theexperiment is to investigate the effect of asoprisnil on endometrial receptivity.Methods: Healthy adult female kunming mice, weighing25±5g, were purchased. Vaginal smearwas taken daily, and only estrus female mice were caged individually with fertile males. Positivemating was verified by identifying copulation plugs in vagina, and designated as Day1ofpregnancy. Post-coitus mice were randomized divided into vehicle control group, low dosage(0.05mg/10g/d), medium dosage (0.1mg/10g/d) and high dosage (0.2mg/10g/d) asoprisnil group.Post-coitus animals were gavaged Asoprisnil administered from Day1to Day3of pregnancy, onceevery day. Animals were sacrificed on Day5of pregnancy, and uteri were sampled. Some of theuteri were fixed by4%neutral paraformaldehyde; hematoxylin and eosin stain was used to evaluatethe morphology changes of endometrium under optical microscope. Immunohistochemistry (IHC)was then used to test the expression of PCNA in endometrium. The other uteri were fixed by2.5%glutaraldehyde, transmission electron microscopy and scanning electron microscopy were used toevaluate the ultrastructure changes of endometrium.Results:①The morphology changes of endometrium in implantation period underoptical microscope: In the vehicle control group, the endometrium of the implantationperiod was characterized by luminal gland proliferation with columnar surface epithelialcells, glands and vascular were congestion and equality scattered among the decidua cellswhich was big and with abundant matrix. While in asoprisnil groups, the gland proliferationwas not significant, the luminal epithelial cells were single and multiple layers distributedirregularly. Stromal cells were compact and small, without decidua transformation.Glandular and vascular was infrequent in endometrium. ②The expression of the PCNA in endometrium in implantation period: IHC indicated thatPCNA was expressed both in luminal epithelium cells and stromal cells. In luminalepithelium cells, the AIOD in vehicle control group, low dose group, medium dose groupand high dose group were respectively as0.21±0.03,0.14±0.02,0.16±0.03and0.15±0.01,the difference were statistically significant when compared with the vehicle control group(P<0.01). while in stromal cells, the AIOD in vehicle control group, low dose group,medium dose group and high dose group were respectively as0.15±0.02,0.17±0.02,0.18±0.03and0.17±0.01, and the difference was statistically significant (P<0.001).③The expression of the pinopodes under scanning electronic microscope: In vehiclecontrol group, the borders of the uterine epithelium cells were clear and microvilli wereabsent from the lining cells. The pinopodes were abundant and well developed as largespongy projections. The endometrium surface was coverd with large amount of developedtpinopodes.While in high dose asoprisnil group, there was a reduced number of maturepinopodes as evidenced by patches of luminal surface dominated by regressing pinopodes.Small tips of microvilli reappeared on the membranes of pinopodes, and the membraneswere significantly wrinkled. The cell size was decreased. However, in low dose asoprisnilgroup, the luminal epithelium cells were slightly bulging and covered with dense microvilli,and they were defined as developing pinopodes.④Ultrastructure change of the implantation period endometrium under transmissionelectron microscope: The transmission electron microscope result indicated that the luminalepithelium was covered with monolayer, neatly arranged columnar cells in vehicle controlgroup. The tight junction between epithelium cells were up-regulated in asoprisnil group,the tight junction was curver and longer in asoprisnil than in vehicle control group. Invehicle control group, there was large number of lipid drops, glycogen and mitochondria oflow electron density in the cytoplasm of the epithelium cells, as well as rough endoplasmic reticulum and Golgi apparatus that indicated active secretory. While in low dosage groupand high dosage asoprisnil group, the luminal epithelium cells were monolayer or multiplelayers. Glycogen and lipids were rare in the cytoplasm of the epithelium cells. Also therewere plenty of mitochondria of high electron density.Conclusion Asoprisnil suppressed the proliferation of the luminal epithelium cells,promoted the proliferation of the stromal cells and obstructed the decidua process of thestromal cells in endometrium of implantation period in mice. Furthermore, asoprisnilinterrupted the development of pinopodes, up-regulated the tight junction between theluminal epithelium. All these morphology and function changes of the pereimplantationendometrium caused the implantation failure in mice. |
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