Espey L L, Tanaka N, Woodard D S, Harper M J, Okamura H
Department of Biology, Trinity University, San Antonio, Texas 78283.
Biol Reprod. 1989 Jul;41(1):104-10. doi: 10.1095/biolreprod41.1.104.
Platelet-activating factor (PAF) is a biologically active phospholipid that is released locally during acute inflammatory reactions and tissue injury. Since there is evidence that the biochemical events of mammalian ovulation resemble an inflammatory reaction, the objective of this study was to determine whether ovarian levels of PAF change during ovulation. At 2-h intervals during the ovulatory process in gonadotropin-primed 25-day-old Wistar rats, the ovaries were extirpated, homogenized, and extracted for lipids. The extracts were subjected to thin-layer chromatography (TLC), and the portion of the silica gel that comigrated with PAF was re-extracted and assayed for PAF activity. The PAF was measured (in fmole equivalents of synthetic PAF) by a bioassay based on the capacity of aliquots of the extracts to release [3H]-serotonin from platelets isolated from whole blood of rabbits and prelabeled with [3H]-serotonin. The ovarian level of PAF decreased (p less than 0.01) by 36% from 6.67 +/- 0.77 to 4.27 +/- 0.45 fmoles/mg ovary by 2 h after treatment with human chorionic gonadotropin (hCG), and it declined another 14% by 4 h after hCG. The ovarian PAF remained at this reduced level for up to 24 h after hCG. The administration of indomethacin (5 mg/rat, s.c.) or epostane (5 mg/rat, s.c.) at 1 h after hCG prevented ovulation, but neither drug affected the decline in ovarian PAF. Preliminary tests showed that the lipid extracts from the ovaries also contained PAF inhibitor(s) that comigrated with PAF on the TLC plates. Similar to PAF, the lipid-soluble inhibitor(s) decreased (p less than 0.05) in the ovaries within 4 h after hCG treatment.(ABSTRACT TRUNCATED AT 250 WORDS)
血小板活化因子(PAF)是一种生物活性磷脂,在急性炎症反应和组织损伤时于局部释放。鉴于有证据表明哺乳动物排卵的生化事件类似于炎症反应,本研究的目的是确定排卵期间卵巢中PAF的水平是否发生变化。在促性腺激素预处理的25日龄Wistar大鼠排卵过程中,每隔2小时将卵巢摘除、匀浆并提取脂质。提取物进行薄层色谱(TLC)分析,与PAF共迁移的硅胶部分重新提取并检测PAF活性。通过生物测定法(基于提取物等分试样从用[³H] - 血清素预标记的兔全血中分离的血小板释放[³H] - 血清素的能力)测量PAF(以合成PAF的飞摩尔当量计)。用人绒毛膜促性腺激素(hCG)处理后2小时,卵巢PAF水平从6.67±0.77飞摩尔/毫克卵巢降至4.27±0.45飞摩尔/毫克卵巢,下降了36%(p<0.01),hCG处理后4小时又下降了14%。hCG处理后长达24小时,卵巢PAF维持在这一降低水平。hCG处理后1小时皮下注射吲哚美辛(5毫克/只大鼠)或依波斯坦(5毫克/只大鼠)可阻止排卵,但两种药物均不影响卵巢PAF的下降。初步试验表明,卵巢脂质提取物中还含有在TLC板上与PAF共迁移的PAF抑制剂。与PAF相似,hCG处理后4小时内,卵巢中脂溶性抑制剂减少(p<0.05)。(摘要截取自250字)