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本文采用羊膜双室培养系统,观察了18甲基炔诺酮(LNG)对猪卵巢颗粒细胞(G-C)和卵泡内膜细胞(T-C)在甾体激素生成过程中的影响。生长在羊膜两侧的G-C和T-C在加入或不加FSH、LH及各种不同浓度LNG的条件下孵育48小时,用RIA测定内、外室培养液中孕酮(P)和雌二醇(E_2)的含量,并与单独培养时的结果相比较。结果表明:①在FSH刺激时,LNG(30、3000nmol/L)明显抑制双室培养的G-C的P和E_2产量:P产量由55.1±3.4μmol/L降为25.8±1.8和20.3±3.8μmol/L;E_2产量由9.35±0.06nmol/L降至5.24±0.64和3.34±0.72nmol/L。但对P和E_2的基础水平无影响。②不论有或无LH刺激,LNG(30、3000nmoL/L)均明显抑制双室培养的T-C的P产量。有LH刺激时,P产量由70.9±6.5μmoL/L分别降为47.1±11.8和4.8±0.5μmol/L;在无LH刺激时,则由26.9±1.7μmol/L分别降至16.9±1.1和5.6±0.9μmol/L。结论:①LNG抑制双室培养中G-C的P和E_2产量,这种抑制作用是通过降低促性腺激素的刺激作用而产生的;②LNG不但抑制T-C的P基础分泌量,而且还表现为降低促性腺激素对P的刺激效应。③双室培养系统模拟了两种卵巢细胞在体时的旁分泌调节作用,与单独培养相比,是研究避孕药对卵巢功能影响的一种更为理想的模型。
In this study, amniotic double chamber culture system was used to observe the effect of 18-norethindrone (LNG) on steroidogenesis in porcine ovarian granulosa cells (G-C) and follicular endometrial cells (T-C) The GCs and TCs grown on both sides of the amnion were incubated for 48 hours with and without FSH, LH and different concentrations of LNG. The contents of progesterone (P) and estradiol E_2) content and compared with the results when cultured alone. The results showed that: (1) At the FSH stimulation, LN (30, 3000 nmol / L) significantly inhibited the P and E 2 production of GC in double chamber culture: P production decreased from 55.1 ± 3.4 μmol / L to 25.8 ± 1.8 and 20.3 ± 3.8 μmol / L; the yield of E_2 decreased from 9.35 ± 0.06nmol / L to 5.24 ± 0.64 and 3.34 ± 0.72nmol / L. But had no effect on the basal levels of P and E_2. ② LN production (30, 3000nmoL / L) significantly inhibited P-production of T-C cultured in double chamber with or without LH stimulation. With LH stimulation, the P production decreased from 70.9 ± 6.5μmol / L to 47.1 ± 11.8 and 4.8 ± 0.5μmol / L, respectively, and from 26.9 ± 1.7μmol / L to 16.9 ± 1.1 and 5.6 ± 0.9 μmol / L. CONCLUSION: (1) LNG inhibits the production of P and E 2 in GC in double-chamber culture. This inhibition is produced by the reduction of gonadotrophin stimulation. (2) LNG not only inhibits the basal secretion of TC but also decreases the expression of gonadotropin Stimulation effect on P. ③ The dual-chamber culture system simulates the paracrine regulation of the two ovarian cells in vivo, which is a more ideal model to study the effect of contraceptives on ovarian function than single culture.