College of Veterinary Medicine, University of Duhok, Kurdistan Regional Government and Scientific Research, Kurdistan, Iraq.
School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicestershire, UK.
J Endocrinol. 2020 Apr;245(1):79-92. doi: 10.1530/JOE-19-0443.
Reproductive tract inflammatory disease (RTID) commonly occurs after the traumatic events of parturition and adversely affects follicular function. This study is the first to describe the cellular and steroidogenic characteristics of corpora lutea from cattle with RTID and the effects of pathogen-associated molecular patterns (PAMPs) on luteal angiogenesis and function in vitro. Luteal weight (P < 0.05) and progesterone content (P < 0.05) were reduced (1.2-fold) in cows with RTID, accompanied by reduced CYP11A (P < 0.05), HSD3B (P < 0.01) and STAR (P < 0.01) protein expression. Immunohistochemistry revealed that luteal vascularity (VWF) and pericyte (ACTA2) coverage were >3-fold lower in RTID cows (P < 0.05). To link these observations to bacterial infection and determine specificity of action, a physiologically relevant luteal angiogenesis culture system examined the effects of PAMPs on endothelial cell (EC) network formation and progesterone production, in the presence of pro-angiogenic factors. Luteal EC networks were reduced ≤95% (P < 0.05) by lipopolysaccharide (LPS, toll-like receptor (TLR) 4 agonist) but not by TLR2 agonists lipoteichoic acid or peptidoglycan. Conversely, progesterone production and steroidogenic protein expression were unaffected by PAMPs (P > 0.05). Moreover, the adverse effect of LPS on luteal EC networks was dose-dependent and effective from 1 ng/mL (P < 0.05), while few EC networks were present above 10 ng/mL LPS (P < 0.001). LPS reduced proliferation (P < 0.05) and increased apoptosis of EC (P < 0.001). The specific TLR4 inhibitor TAK242 reversed the effects of LPS on EC networks. In conclusion, luteal vasculature is adversely sensitive to LPS acting via TLR4, therefore ovarian exposure to LPS from any Gram-negative bacterial infection will profoundly influence subsequent reproductive potential.
生殖道炎症性疾病（RTID）常发生于分娩创伤后，会对卵泡功能产生不良影响。本研究首次描述了患有 RTID 的牛的黄体的细胞和类固醇生成特征，以及病原体相关分子模式（PAMPs）对黄体血管生成和体外功能的影响。患有 RTID 的奶牛的黄体重量（P < 0.05）和孕酮含量（P < 0.05）降低（降低 1.2 倍），同时 CYP11A（P < 0.05）、HSD3B（P < 0.01）和 STAR（P < 0.01）蛋白表达降低。免疫组织化学显示，RTID 奶牛的黄体血管性（VWF）和周细胞（ACTA2）覆盖率降低了 >3 倍（P < 0.05）。为了将这些观察结果与细菌感染联系起来并确定作用的特异性，采用生理相关的黄体血管生成培养系统研究了 PAMPs 在存在促血管生成因子的情况下对内皮细胞（EC）网络形成和孕酮产生的影响。脂多糖（LPS，Toll 样受体（TLR）4 激动剂）可使黄体 EC 网络减少 ≤95%（P < 0.05），但 TLR2 激动剂脂磷壁酸或肽聚糖则不会。相反，PAMPs 对孕酮产生和类固醇生成蛋白表达没有影响（P > 0.05）。此外，LPS 对黄体 EC 网络的不良影响呈剂量依赖性，1ng/mL 时即可起效（P < 0.05），而 LPS 浓度高于 10ng/mL 时则很少有 EC 网络存在（P < 0.001）。LPS 降低了 EC 的增殖（P < 0.05）并增加了其凋亡（P < 0.001）。特异性 TLR4 抑制剂 TAK242 逆转了 LPS 对 EC 网络的影响。总之，黄体血管系统对 LPS 敏感，LPS 通过 TLR4 发挥作用，因此，卵巢暴露于任何革兰氏阴性菌感染的 LPS，将极大地影响随后的生殖潜力。
