猪子宫内膜肥大细胞消涨规律初探

雌性小型猪１５头，分为二月龄、间情期、民情前期、发情期、妊娠期等六组。颈动脉放血处死，取子宫组织，经甲苯胺改良法和免疫组化法染色，观察肥大细胞的动态变化。子宫内膜肥大细胞的数量随个体的生长而增加。在发情周期中肥大细胞的数量呈规律性的变化：在民情前期一时性地剧增，至发情期陡然降至显著低于间情期的水平，在妊娠期也维持低于间情期的水平。本实验的结果提示，子宫内膜肥大细胞的数量可能反映子宫局部的免疫水平。     

脂肪酸结合蛋白4在小鼠子宫蜕膜中的表达

[目的]检测脂肪酸结合蛋白4(FABP4)在小鼠子宫蜕膜细胞中的表达。[方法]获取正常的小鼠子宫蜕膜组织和人工诱导蜕膜化组织,同时利用酶消化分离小鼠子宫内膜基质细胞,进行体外诱导蜕膜化,利用实时定量PCR检测脂肪酸结合蛋白4在子宫蜕膜组织细胞中的表达。[结果]FABP 4在小鼠正常子宫蜕膜组织、人工诱导蜕膜化组织及子宫内膜蜕膜化基质细胞中的表达水平均显著高于对照组。[结论]FABP4可能参与了小鼠子宫内膜蜕膜化过程,为研究蜕膜化机理奠定了基础。     

Comparative analysis of proteomic profiles between endometrial caruncular and intercaruncular areas in ewes during the peri-implantation period

The endometrium of sheep consists of plenty of raised aglandular areas called caruncular (C), and intensely glandular intercaruncular areas (IC). In order to better understand the endometrium involved mechanisms of implantation, we used LC-MS/MS technique to profile the proteome of ovine endometrial C areas and IC areas separately during the peri-implantation period, and then compared the proteomic profiles between these two areas. We successfully detected 1740 and 1813 proteins in C areas and IC areas respectively. By comparing the proteome of these two areas, we found 170 differentially expressed proteins (DEPs) (P < 0.05), functional bioinformatics analysis showed these DEPs were mainly involved in growth and remodeling of endometrial tissue, cell adhesion and protein transport, and so on. Our study, for the first time, provided a proteomic reference for elucidating the differences between C and IC areas, as an integrated function unit respectively, during the peri-implantation period. The results could help us to better understand the implantation in the ewes. In addition, we established a relatively detailed protein database of ovine endometrium, which provide a unique reference for further studies.     

Effect of acupuncture on CD4+ CD25+ Foxp3+ regulatory T-cells in rats of embryo implantation failure

AIM：To observe the effect of acupuncture on CD4+ CD25+ Foxp3(forkhead box P3)+ regulatory T-cells(Treg cells) in rats with embryo implantation failure. METHODS：One hundred and forty-four pregnant rats were randomly divided into control group(N), mifepristone treatment group(M), mifepristone+acupuncture treatment group(A) and mifepristone+progestin treatment group(W). The rats in groups M, A and W were treated with mifepristone-sesame oil solution on day 1, while the rats in group N were injected with the same amount of sesame oil. The Housanli(ST36) and Sanyinjiao(SP6) points were selected for acupuncture. From day 1 to the time of death, the rats in group A were fasten up and then the acupuncture was performed. Accordingly, the rats in group N and group M were only fixed, and the rats in group W were given progestin. Implanted embryos in each group were counted. The proportions of CD4+ CD25+ Foxp3+ Treg cells in peripheral blood and CD4+ Foxp3+ Treg cells in the endometrium were detected by flow cytometry. The mRNA and protein levels of Foxp3 were determined by real-time PCR and Western blotting, respectively. RESULTS：Compared with group N, the number of implanted embryos, the percentages of CD4+ CD25+ Foxp3+ Treg cells in peripheral blood and CD4+ Foxp3+ Treg cells in the endometrium, and the expression of Foxp3 protein and mRNA in the endometrium were significantly decreased in group M(P<005). Compared with group M, the above indexes in group A and group W were significantly increased. CONCLUSION: The effect of acupuncture in rats with embryo implantation failure may be closely correlated with the modulation of CD4+ CD25+ Foxp3+ Treg cells.     

Comparative analysis of proteomic profiles between endometrial caruncular and intercaruncular areas in ewes during the peri-implantation period

The endometrium of sheep consists of plenty of raised intercaruncular areas （IC）. In order to better understand aglandular areas called caruncular （C）, and intensely glandular the endometrium involved mechanisms of implantation, we used LC-MS/MS technique to profile the proteome of ovine endometrial C areas and IC areas separately during the peri-implantation period, and then compared the proteomic profiles between these two areas. We successfully detected 1740 and 1813 proteins in C areas and IC areas respectively. By comparing the proteome of these two areas, we found 170 differentially expressed proteins （DEPs） （P 〈 0.05）, functional bioinformatics analysis showed these DEPs were mainly involved in growth and remodeling of endometrial tissue, cell adhesion and protein transport, and so on Our study, for the first time, provided a proteomic reference for elucidating the differences between C and IC areas, as an integrated function unit respectively, during the peri-implantation period. The results could help us to better understand the implantation in the ewes. In addition, we established a relatively detailed protein database of ovine endometrium, which provide a unique reference for further studies.     

Non-genomic actions of progesterone and estrogens in regulating reproductive events in domestic animals

It has been established that nuclear receptors mediate the action of estrogens and progestins in regulating gene expression in the hypothalamic-hypophyseal-gonadal axis of domestic animals during various reproductive states. Results of recent in vitro studies suggest that estradiol-17beta and progesterone can act non-genomically to affect signal transduction responses in target cells by binding to receptors in the plasma membrane. The genomic action of steroids is generally detectable in hours to days whereas non-genomic responses of cells occur in seconds to minutes. The nature of the plasma membrane receptors for estrogens and progesterone has been explored but has not been conclusively established for all cell types studied. In the ewe, estradiol-17beta or estradiol-bovine serum albumin conjugate has been shown by in vitro and in vivo approaches to act non-genomically to suppress luteinizing hormone secretion by gonadotropes and stimulate production of nitric oxide by uterine arterial endothelial cells. Progesterone has been shown to inhibit oxytocin (OT) binding to its receptor in isolated ovine endometrial plasma membranes. This non-genomic action of progesterone blocks OT activation of the phosphoinositide cascade and production of prostaglandin F(2alpha) by ovine and bovine endometrium. The acrosome reaction of caprine and porcine spermatozoa is activated by the non-genomic action of progesterone. Further research is required to define the biological significances of the non-genomic actions of estrogens and progestins.     

The Effect of Cysteine-Rich Secretory Protein-3 and Lactoferrin on Endometrial Cytokine mRNA Expression After Breeding in the Horse

The equine uterus undergoes a transient innate immune response after breeding, also known as mating-induced endometritis. The deposition of spermatozoa triggers the expression of pro-inflammatory cytokines, which results in the migration of polymorphonuclear neutrophils (PMNs) into the endometrium and the uterine lumen. Select seminal plasma proteins, specifically cysteine-rich secretory protein 3 (CRISP-3) and lactoferrin, have been shown to affect the activity of the PMNs, either by suppressing (CRISP-3) or promoting (lactoferrin) the phagocytosis of spermatozoa based on their viability in vitro. Conjointly, many components of inseminate, including seminal plasma, bacteria, and spermatozoa itself, have shown to have an effect on the expression of endometrial cytokines after breeding. The objective of this study was to determine if select proteins affect the mRNA expression of endometrial cytokines after insemination. Six mares were bred during four consecutive estrous cycles with treatments in randomized order of: 1mg/mL CRISP-3, 150 ug/mL lactoferrin, seminal plasma, or Lactated Ringer’s Solution (LRS) to a total volume of 10 mL combined with 1×10⁹ progressively motile spermatozoa pooled from two stallions. Six hours after treatment, an endometrial biopsy was obtained for qPCR analysis. No treatment effects were found for the mRNA expression of IL-1β, IL-6, IL-8, IL-10, TNFα, and IFNγ, while lactoferrin significantly suppressed the mRNA expression of IL-1RN when compared to LRS. In conclusion, the seminal plasma proteins CRISP-3 and lactoferrin have minimal effect on the expression of select endometrial cytokines at 6 hours post breeding.