Jacobs A L, Sehgal P B, Julian J, Carson D D
Department of Biochemistry and Molecular Biology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Endocrinology. 1992 Sep;131(3):1037-46. doi: 10.1210/endo.131.3.1505448.
Uterine stromal (USC) and uterine epithelial (UEC) cells were isolated from immature and mature mice to determine their ability to secrete interleukin-6 (IL-6) in response to ovarian steroids, IL-1 alpha, and soluble products produced by the heterologous cell type. In addition, the effect of IL-6 on embryo attachment and outgrowth in vitro was determined. UEC cultured on nitrocellulose filter inserts in a polarized manner secreted IL-6 with a 2.5- to 5-fold apical vs. basal preference, as determined by a B9 hybridoma cell proliferation assay and enzyme-linked immunosorbent assay. The hormonal status of animals at the time uteri were removed did not influence subsequent secretion of IL-6, as UEC isolated from immature, diestrous, and estrous stage mice exhibited both a similar amount and had a similar apical preference for secretion of IL-6. The addition of 17 beta-estradiol (E) to UEC cultures markedly inhibited total IL-6 secretion, but did not affect vectorial secretion. The inhibitory effect of E on IL-6 secretion by UEC was consistent with an apparent decrease in IL-6 transcript observed by a reverse transcriptase polymerase chain reaction assay. Other transcripts detected by this assay in UEC included IL-1 alpha, but not IL-1 beta or tumor necrosis factor-alpha. Secretion of IL-6 by UEC was not stimulated by IL-1 alpha, conditioned medium from USC, or coculture with USC. USC secreted IL-6, and while this also was inhibited by E, progesterone was more effective in this regard at physiological concentrations. In addition, there was a synergistic effect of E plus progesterone on inhibition of IL-6 secretion by USC. Secretion of IL-6 by USC was stimulated by IL-1 alpha, and coculture studies demonstrated the ability of UEC to stimulate a several-fold increase in IL-6 secretion by USC. The cytokine transcripts detected in USC cultures included IL-6 and IL-1 alpha, but not IL-1 beta. Transcripts for tumor necrosis factor-alpha were present in USC only after culture with IL-1 alpha. IL-6 added to blastocysts on laminin-coated tissue culture wells resulted in a transient inhibition of the rate of blastocyst attachment and, to a greater extent, an inhibition of the rate of embryo outgrowth. In addition, IL-6 inhibited the size of embryo outgrowths at 24 and 48 h of culture.(ABSTRACT TRUNCATED AT 400 WORDS)
从小鼠的未成熟和成熟阶段分离出子宫基质细胞（USC）和子宫上皮细胞（UEC），以确定它们对卵巢类固醇、白细胞介素-1α（IL-1α）以及异源细胞类型产生的可溶性产物作出反应时分泌白细胞介素-6（IL-6）的能力。此外，还确定了IL-6对体外胚胎着床和生长的影响。通过B9杂交瘤细胞增殖试验和酶联免疫吸附试验测定，以极化方式培养在硝酸纤维素滤膜插入物上的UEC分泌IL-6时，其顶端分泌量是基底分泌量的2.5至5倍。去除子宫时动物的激素状态并不影响随后IL-6的分泌，因为从未成熟、动情间期和动情期小鼠分离出的UEC分泌的IL-6量相似，且顶端分泌偏好也相似。向UEC培养物中添加17β-雌二醇（E）可显著抑制IL-6的总分泌，但不影响其定向分泌。E对UEC分泌IL-6的抑制作用与逆转录聚合酶链反应试验观察到的IL-6转录本明显减少一致。该试验在UEC中检测到的其他转录本包括IL-1α，但不包括IL-1β或肿瘤坏死因子-α。UEC分泌IL-6不受IL-1α、USC的条件培养基或与USC共培养的刺激。USC分泌IL-6，虽然这也受E抑制，但在生理浓度下孕酮在这方面更有效。此外，E加孕酮对USC分泌IL-6有协同抑制作用。USC分泌IL-6受IL-1α刺激，共培养研究表明UEC有能力刺激USC的IL-6分泌增加几倍。在USC培养物中检测到的细胞因子转录本包括IL-Il-6和IL-1α，但不包括IL-1β。仅在与IL-1α共培养后，USC中才出现肿瘤坏死因子-α的转录本。添加到包被层粘连蛋白的组织培养孔中的囊胚上的IL-6导致囊胚附着率短暂降低，并且在更大程度上抑制胚胎生长率。此外，IL-6在培养24小时和48小时时抑制胚胎生长的大小。（摘要截短于400字）
