目的:探讨促性腺激素预处理的小鼠排卵过程中ADAMTS-1在卵巢中的表达与排卵的关系。方法:将22~24天大小的C57/BL6雌性小鼠随机分成10组,分别为孕马血清促性腺激素(PMSG)组(D1组)、注射hCG后2h、4h、8h、12h、16h、24h、48h、72h组(D2~D9组)及对照组(DO组)。采用原位杂交进行组织学定位,逆转录聚合酶链反应(RT-PCR)进行半定量,在mRNA水平上检测排卵过程中ADAMTS-1在小鼠卵巢中的动态表达。结果:DO、D1、D8、D9组均无ADAMTS-1的表达,132-177组均有ADAMTS-1的表达,主要表达于卵巢表面大卵泡的颗粒细胞。注射hCG后2~12h卵巢ADAMTS-1mRNA的表达逐渐增强,注射hCG后12h达到高峰,其ADAMTS-1mRNA表达的相对值为(1.357±0.328),与各组比较均有极显著差异(P〈0.05),此后表达逐渐下降,至注射hCG后48h无表达,但D2、D3、D4、D6、D7组之间无显著性差异(P〉0.05)。结论:ADATMS-1可能是排卵过程中重要的信号传导分子。
Objective: To detect ovarian expression of a relatively new type of disintegrin and metalloproteinase with throm- bospondin motifs ( ADAMTS - 1 ) during ovulation in the gonadotropin - primed immature mice. Methods: Immature C57/BL6 mice were primed with 4IU PMSG, and the temporal pattern of expression of the ADAMTS - 1 gene was delineated by extracting ovarian RNA at 0, 2, 4, 8, 12, 16, 24, 48 and 72 h after induction of ovulation by injecting the primed animals with 5 IU hCG. Forty mice were randomly allocated into 10 groups of 4 mice. In situ hybridization was used to locate the ADAMTS - 1 mRNA epression and reverse transcription - polymerase chain reaction (RT- PCR) was used to semiquantitatively detect the expression of ADAMTS -1 mRNA during ovulation. Results: In situ hybridization micrographs showed that the ADAMTS - 1 mRNA was expressed primarily in the granulosa layer of the larger follicles around the periphery of the ovarian mass. In contrast, the gene was not expressed in smaller follicles located toward the center of the ovaries, or in atretic follicles. ADAMTS - 1 was not detected at 0, 48 and 72 h after hCG administration and the control group. The RT - PCR data and in situ hybridization micrographs all showed that the expression of ADAMTS - 1 gene increased gradually from 2 h after hCG and reached a peak at 12 h after hCG. The relative quantity of ADAMTS -1 mRNA expression at 12 h after hCG was 1. 357±0. 328, significantly higher than all the other groups (P 〈0. 05) . The expression declined later. But there was not a significant difference among 2, 4, 8, 16, 24 h after hCG (P 〉 0. 05 ) . Conclusion: ADAMTS -1 probably plays an important role in signal transduction during ovulation.