Pro-inflammatory cytokines secreted by macrophages and other cell types are implicated as intraovarian factors affecting different aspects of ovarian function including follicle and corpus luteum 'turnover', steroidogenesis and angiogenesis. Here, we compared granulosal (GC) and thecal (TC) expression of TNF, IL6 and their receptors (TNFRSF1A, TNFRSF1B and IL6R) during bovine antral follicle development; all five mRNA transcripts were detected in both GC and TC and statistically significant cell-type and follicle stage-related differences were evident. Since few studies have examined cytokine actions on TC steroidogenesis, we cultured TC under conditions that retain a non-luteinized 'follicular' phenotype and treated them with TNFα and IL6 under basal and LH-stimulated conditions. Both TNFα and IL6 suppressed androgen secretion concomitantly with CYP17A1 and LHCGR mRNA expression. In addition, TNFα reduced INSL3, HSD3B1 and NOS3 expression but increased NOS2 expression. IL6 also reduced LHCGR and STAR expression but did not affect HSD3B1, INSL3, NOS2 or NOS3 expression. As macrophages are a prominent source of these cytokines in vivo, we next co-cultured TC with macrophages and observed an abolition of LH-induced androgen production accompanied by a reduction in CYP17A1, INSL3, LHCGR, STAR, CYP11A1 and HSD3B1 expression. Exposure of TC to bacterial lipopolysaccharide also blocked LH-induced androgen secretion, an effect reduced by a toll-like receptor blocker (TAK242). Collectively, the results support an inhibitory action of macrophages on thecal androgen production, likely mediated by their secretion of pro-inflammatory cytokines that downregulate the expression of LHCGR, CYP17A1 and INSL3. Bovine theca interna cells can also detect and respond directly to lipopolysaccharide.

译文

巨噬细胞和其他细胞类型分泌的促炎细胞因子被认为是影响卵巢功能不同方面的卵巢内因素,包括卵泡和黄体 “周转”,类固醇生成和血管生成。在这里,我们比较了牛窦卵泡发育过程中TNF,IL6及其受体 (TNFRSF1A,TNFRSF1B和IL6R) 的颗粒 (GC) 和thecal (TC) 表达; 在GC和TC中均检测到所有五个mRNA转录本,并且具有统计学意义的细胞类型和卵泡阶段相关差异。由于很少有研究检查细胞因子对TC类固醇生成的作用,因此我们在保留非黄体化 “卵泡” 表型的条件下培养TC,并在基础和LH刺激的条件下用tnf α 和IL6处理它们。Tnf α 和IL6均与CYP17A1和LHCGR mRNA表达同时抑制雄激素分泌。此外,tnf α 降低了INSL3,HSD3B1和NOS3的表达,但增加了NOS2的表达。IL6还降低了LHCGR和STAR的表达,但不影响HSD3B1,INSL3,NOS2或NOS3的表达。由于巨噬细胞是体内这些细胞因子的主要来源,我们接下来将TC与巨噬细胞共培养,并观察到LH诱导的雄激素产生的消除,伴随着CYP17A1,INSL3,LHCGR,STAR,CYP11A1和HSD3B1表达的降低。将TC暴露于细菌脂多糖中也会阻止LH诱导的雄激素分泌,这种作用被toll样受体阻滞剂 (TAK242) 降低。总的来说,结果支持巨噬细胞对鞘雄激素产生的抑制作用,可能是由其分泌促炎细胞因子介导的,这些细胞因子下调了LHCGR,CYP17A1和insl3的表达。牛卵泡膜细胞也可以检测脂多糖并直接响应。

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