Buhi W C, Alvarez I M, Pickard A R, McIntush E W, Kouba A J, Ashworth C J, Smith M F
Department of Obstetrics and Gynecology, University of Florida College of Medicine, Gainesville 32610-0294, USA.
Biol Reprod. 1997 Jul;57(1):7-15. doi: 10.1095/biolreprod57.1.7.
It has been suggested that tissue inhibitor of metalloproteinases (TIMP)-1 has a role in reproductive tissues, regulating tissue remodeling or enhancing embryonic development. Oviductal TIMP-1 mRNA levels and protein expression were examined in gilts during the estrous cycle and early pregnancy and in steroid-treated ovariectomized (OVX) gilts by explant culture, two-dimensional SDS-PAGE and fluorography, dot-blot hybridization, immunoblot analysis, RIA, and immunocytochemical studies. TIMP-1 mRNA levels in the oviduct during the estrous cycle were greater (p < 0.02) on Days 2, 15, and 18 than on other days examined, and analysis of oviductal functional segments indicated an effect of day (p < 0.003), an effect of segment (p < 0.007), and a day x segment effect (p < 0.03). The level of TIMP-1 mRNA was greater (p < 0.003) in the isthmus (I) on Day 2 than in the ampulla (A) or infundibulum (INF) or on other days examined (0 and 12). In steroid-treated OVX gilts, an effect of treatment with estradiol valerate (EV) + progesterone (P4) was shown with increased (p < 0.003) TIMP-1 mRNA levels. De novo synthesis of TIMP-1 protein was found throughout the estrous cycle and early pregnancy in all functional segments, but protein expression was greater in the I and greatest on Day 2. In steroid-treated OVX gilts, TIMP-1 protein synthesis was greatest in the I regardless of treatment, but with increased intensity after EV+P4 treatment. TIMP-1 protein was found in oviductal flushings during the estrous cycle and early pregnancy, and in steroid-treated OVX gilts regardless of day, status, or treatment. Differences in TIMP-1 concentrations in oviductal fluid were found by day (p < 0.001), with breed differences detected between the Meishan and standard Western breeds. TIMP-1 protein was immunolocalized primarily to luminal epithelium of the INF, A, and I on all days of the estrous cycle and early pregnancy and to some cells in the stroma and blood vessel walls. Staining intensity correlated with TIMP-1 protein levels in oviductal flushings. The role of TIMP-1 in the oviduct remains to be established.
有人提出金属蛋白酶组织抑制剂（TIMP）-1在生殖组织中发挥作用，调节组织重塑或促进胚胎发育。通过外植体培养、二维SDS-PAGE和荧光成像、斑点印迹杂交、免疫印迹分析、放射免疫分析和免疫细胞化学研究，检测了发情周期和妊娠早期后备母猪以及经类固醇处理的去卵巢（OVX）后备母猪输卵管中TIMP-1 mRNA水平和蛋白表达。发情周期中，输卵管TIMP-1 mRNA水平在第2、15和18天高于其他检测日（p<0.02），对输卵管功能段的分析表明存在日效应（p<0.003）、段效应（p<0.007）和日×段效应（p<0.03）。第2天峡部（I）的TIMP-1 mRNA水平高于壶腹部（A）或漏斗部（INF）或其他检测日（0和12）（p<0.003）。在经类固醇处理的OVX后备母猪中，戊酸雌二醇（EV）+孕酮（P4）处理显示出TIMP-1 mRNA水平升高（p<0.003）。在发情周期和妊娠早期的所有功能段均发现了TIMP-1蛋白的从头合成，但峡部的蛋白表达更高，且在第2天最高。在经类固醇处理的OVX后备母猪中，无论处理如何，峡部的TIMP-1蛋白合成最高，但经EV+P4处理后强度增加。在发情周期和妊娠早期的输卵管冲洗液中以及经类固醇处理的OVX后备母猪中，无论日、状态或处理如何，均发现了TIMP-1蛋白。输卵管液中TIMP-1浓度存在日差异（p<0.001），梅山猪和标准西方品种之间存在品种差异。在发情周期和妊娠早期的所有日子里，TIMP-1蛋白主要免疫定位在漏斗部、壶腹部和峡部的腔上皮以及基质和血管壁的一些细胞中。染色强度与输卵管冲洗液中TIMP-1蛋白水平相关。TIMP-1在输卵管中的作用仍有待确定。
