Chang C C, Hsieh Y Y, Chung J G, Tsai H D, Tsai C H
Department of Obstetrics and Gynecology, China Medical College Hospital, Taichung, Taiwan.
J Assist Reprod Genet. 2001 Dec;18(12):660-4. doi: 10.1023/a:1013167317997.
To evaluate the activities of acetyl coenzyme A (AcCoA):arylamine N-acetyltransferase (NAT) of intact cumulus granulosa cells and the role of leukemia inhibitory factor (LIF) upon their NAT activities.
Thirty women accepted controlled ovarian hyperstimulation (COH) and oocyte retrievals. Human cumulus granulosa cells were obtained during oocyte retrievals. Using 2-aminofluorene (2-AF) and p-aminobenzoic acid (PABA) as substrates, NAT activity of all samples was determined by high pressure liquid chromatography. After the incubation with different time and concentrations of 2-AF, PABA, and LIF, 2-acetyl-aminofluorene (2-AAF) and N-acetyl-PABA (N-Ac-PABA) were measured.
After incubation with 2.812, 5.625, 11.25, and 22.5 microM of 2-AF/PABA, their product concentrations (2-AAF/N-Ac-PABA) were 0.42/0.32, 0.76/0.58, 1.29/1.04, and 1.94/1.26 nmol/10(6) cells, respectively. After 6, 12, 18, and 24 h incubation with 11.25 microM of 2-AF/PABA, their product concentrations were 0.19/0.12, 0.56/0.4, 0.98/0.79, and 1.3/1.0 nmol/10(6) cells, respectively. After incubation with 0, 5, and 50 microM of LIF, the 2-AAF/N-Ac-PABA concentrations were 0.98/0.80, 0.70/0.52, and 0.49/0.30 nmol/10(6) cells, respectively.
Intact human cumulus granulosa cells could acetylate arylamine carcinogen (2-AF) and noncarcinogens drug (PABA). LIF decreased the NAT activities. It provides a model for monitoring the effects of COH and LIF upon the oocytes.
评估完整的卵丘颗粒细胞中乙酰辅酶A(AcCoA):芳胺N - 乙酰基转移酶(NAT)的活性,以及白血病抑制因子(LIF)对其NAT活性的作用。
30名女性接受控制性卵巢刺激(COH)并进行卵母细胞采集。在卵母细胞采集过程中获取人卵丘颗粒细胞。以2 - 氨基芴(2 - AF)和对氨基苯甲酸(PABA)为底物,通过高压液相色谱法测定所有样本的NAT活性。在用不同时间和浓度的2 - AF、PABA和LIF孵育后,测定2 - 乙酰氨基芴(2 - AAF)和N - 乙酰 - PABA(N - Ac - PABA)。
用2.812、5.625、11.25和22.5微摩尔的2 - AF/PABA孵育后,其产物浓度(2 - AAF/N - Ac - PABA)分别为0.42/0.32、0.76/0.58、1.29/1.04和1.94/1.26纳摩尔/10⁶个细胞。用11.25微摩尔的2 - AF/PABA孵育6、12、18和24小时后,其产物浓度分别为0.19/0.12、0.56/0.4、0.98/0.79和1.3/1.0纳摩尔/10⁶个细胞。用0、5和50微摩尔的LIF孵育后,2 - AAF/N - Ac - PABA浓度分别为0.98/0.80、0.70/0.52和0.49/0.30纳摩尔/10⁶个细胞。
完整的人卵丘颗粒细胞能够使芳胺致癌物(2 - AF)和非致癌物药物(PABA)乙酰化。LIF降低了NAT活性。它为监测COH和LIF对卵母细胞的影响提供了一个模型。