Korzekwa A J, Acosta T J, Miklewicz M, Okuda K, Lee S H, Skarzynski D J
Department of Reproductive Immunology and Pathology of the Polish Academy of Sciences, Olsztyn, Poland.
Reprod Domest Anim. 2010 Dec;45(6):e288-96. doi: 10.1111/j.1439-0531.2009.01559.x.
The aim of this study was to determine which cells are the source of production and target for leukotriene (LTs) action within the bovine ovary. Luteal (CL, days 14-16 of the oestrous cycle), steroidogenic cells (LSC) and endothelial cells (LEC) of the bovine corpus luteum (CL), and granulosa cells (GC) were isolated enzymatically, cultured in a monolayer and incubated with LTC(4), LTB(4), Azelastine (an antagonist of LTC(4)) or Dapsone (an antagonist of LTB(4)). Then cells were collected for determination of mRNA expression for LT receptors (LTRs) and 5-lipoxygenase (5-LO) by real time RT-PCR, and media were collected for determination of prostaglandin (PG)E(2), F(2α), progesterone (P4; LSC only), endothelin-1 (ET-1; LEC only) and 17-β oestradiol (E2; GC only). The greatest mRNA expression for LTR-II and 5-LO were found in LEC, whereas LTR-I mRNA expression did not differ among cell types. The level of PGE(2) increased after LTs treatment in each type of ovarian cell, excluding LTC(4) treatment in LEC. The secretion of PGF(2α) was also increased by LTs, but decreased after LTB(4) treatment of LSC. In GC cultures, both LTs stimulated E2 secretion; in LEC cultures, LTB(4) stimulated whereas LTC(4) inhibited P4 secretion; in LEC cultures, LTC(4) stimulated but LTB(4) inhibited ET-1 secretion. The results show that LTs are produced locally and are involved in PGs production/secretion in all examined cells (LSC, LEC and GC) of bovine ovary. Leukotriene treatment modulate secretion of E2, by GC, P4 by LSC and ET-1 by LEC, which indicates that LTs are involved in regulation of ovarian secretory functions.
本研究的目的是确定牛卵巢内白三烯（LTs）产生的细胞来源以及其作用的靶细胞。牛黄体（CL，发情周期第14 - 16天）的黄体细胞（CL）、类固醇生成细胞（LSC）和内皮细胞（LEC），以及颗粒细胞（GC）通过酶解法分离，进行单层培养，并分别与白三烯C4（LTC4）、白三烯B4（LTB4）、氮卓斯汀（LTC4拮抗剂）或氨苯砜（LTB4拮抗剂）孵育。然后收集细胞，通过实时逆转录聚合酶链反应（RT-PCR）测定白三烯受体（LTRs）和5-脂氧合酶（5-LO）的mRNA表达；收集培养基，测定前列腺素（PG）E2、F2α、孕酮（P4，仅针对LSC）、内皮素-1（ET-1，仅针对LEC）和17-β雌二醇（E2，仅针对GC）。结果发现，LEC中LTR-II和5-LO的mRNA表达最高，而LTR-I的mRNA表达在不同细胞类型之间无差异。除LEC中LTC4处理外，每种卵巢细胞类型经LTs处理后PGE2水平均升高。LTs也使PGF2α的分泌增加，但LTB4处理LSC后PGF2α分泌减少。在GC培养物中，两种LTs均刺激E2分泌；在LEC培养物中，LTB4刺激而LTC4抑制P4分泌；在LEC培养物中，LTC4刺激但LTB4抑制ET-1分泌。结果表明，LTs在牛卵巢内局部产生，并参与所有检测细胞（LSC、LEC和GC）中PGs的产生/分泌。白三烯处理可调节GC分泌E2、LSC分泌P4以及LEC分泌ET-1，这表明LTs参与卵巢分泌功能的调节。
