Ito M, Imaizumi H, Hayakawa S, Katou K, Nishiyama Y, Sugiyama Y, Nose Y
Nihon Naibunpi Gakkai Zasshi. 1983 Jul 20;59(7):993-1003. doi: 10.1507/endocrine1927.59.7_993.
We examined the localization of five pregnancy-associated proteins such as pregnancy specific beta 1-glycoprotein (SP1), human chorionic gonadotropin (HCG), human placental lactogen (HPL), alpha 2-PA-glycoprotein (SP3) and pregnancy-associated plasma protein-A (PAPP-A) in the placentae of early and term pregnancies by means of the PAP method in immunohistochemical technology. It was found that the degree of staining of these proteins did not reflect their concentration in serum at each gestational age. Therefore, the concentrations of these proteins in the solution extracted from the placenta were measured. 1) Placental tissues were fixed with a 10% neutral formaldehyde solution and these were embedded into paraffin blocks. These specimens were used for the PAP method. The localizations of five pregnancy-associated proteins from 10 placentae at the gestational age of 7 to 10 weeks were compared with those at the gestation age of 38 to 41 weeks from 7 patients. The staining degree of SP1 in free villi was pale in both early and term pregnancies. HCG was stained deeply in early pregnancy but pale at term pregnancy. HPL was stained deeply at both gestational ages. Horseradish peroxidase reaction products from SP1, HCG and HPL were located chiefly in the syncytiotrophoblast. SP3 was not stained on the placental tissue. PAPP-A was stained to a greater extent in the cytotrophoblast in early pregnancy but pale in the syncytiotrophoblast at term pregnancy. 2) The concentrations of SP1, HCG, HPL, SP3 and PAPP-A were measured in the placental tissues in both early and term pregnancies. Placental tissues were obtained from 24 normal pregnancy patients aborted artificially at 8 to 11 weeks gestation and from 28 patients terminated by normal deliveries at 38 to 40 weeks gestation. The tissue was homogenated with 3 volumes of 1/2 PBS (0.005M phosphate-buffered 0.07M sodium chloride, pH 7.4). The supernatant was removed after centrifugation and stored at -20 degrees C until assayed. The concentration of SP1 was measured by single radial immunodiffusion. That of HCG was assayed by a directed Latex agglutination test (Gestate Slide Eiken). The concentrations of HPL, SP3 and PAPP-A were quantified by rocket immunoelectrophoresis. No significant difference in SP1 levels was shown between early and term pregnancies, and the SP1 level was 4.3 +/- 1.3 (mean +/- 1 S.D.) mg/dl at term pregnancy. The HCG level was 1,100 +/- 300 IU/ml in early pregnancy and at least 20-fold higher than that at term pregnancy.(ABSTRACT TRUNCATED AT 400 WORDS)
我们采用免疫组织化学技术中的PAP法,研究了5种妊娠相关蛋白,即妊娠特异性β1 -糖蛋白(SP1)、人绒毛膜促性腺激素(HCG)、人胎盘催乳素(HPL)、α2 - PA -糖蛋白(SP3)和妊娠相关血浆蛋白 - A(PAPP - A)在早孕和足月妊娠胎盘组织中的定位。结果发现,这些蛋白的染色程度并不能反映其在各孕周血清中的浓度。因此,我们对从胎盘中提取的溶液中这些蛋白的浓度进行了测定。1)胎盘组织用10%中性甲醛溶液固定后,包埋于石蜡块中。这些标本用于PAP法检测。将10例孕7至10周胎盘组织中5种妊娠相关蛋白的定位,与7例孕38至41周胎盘组织中的定位进行了比较。SP1在游离绒毛中的染色程度在早孕和足月妊娠时均较淡。HCG在早孕时染色深,但在足月妊娠时染色淡。HPL在两个孕周时染色均深。SP1、HCG和HPL的辣根过氧化物酶反应产物主要位于合体滋养层。SP3在胎盘组织上未染色。PAPP - A在早孕时细胞滋养层中的染色程度较高,但在足月妊娠时合体滋养层中的染色淡。2)测定了早孕和足月妊娠胎盘组织中SP1、HCG、HPL、SP3和PAPP - A的浓度。胎盘组织取自24例妊娠8至11周人工流产的正常孕妇以及28例妊娠38至40周正常分娩的孕妇。将组织用3倍体积的1/2 PBS(0.005M磷酸盐缓冲的0.07M氯化钠,pH 7.4)匀浆。离心后取上清液,于 - 20℃保存直至检测。SP1的浓度通过单向免疫扩散法测定。HCG的浓度通过乳胶凝集试验(Gestate Slide Eiken)检测。HPL、SP3和PAPP - A的浓度通过火箭免疫电泳法定量。早孕和足月妊娠之间SP1水平无显著差异,足月妊娠时SP1水平为4.3±1.3(均值±1标准差)mg/dl。早孕时HCG水平为1100±300 IU/ml,至少比足月妊娠时高20倍。(摘要截断于400字)