Chapwanya Aspinas, Meade Kieran G, Doherty Michael L, Callanan John J, Mee John F, O'Farrelly Cliona
Comparative Immunology Group, School of Biochemistry and Immunology, Trinity College, Dublin 2, Ireland.
Theriogenology. 2009 Jun;71(9):1396-407. doi: 10.1016/j.theriogenology.2009.01.006. Epub 2009 Feb 23.
Bovine uterine disease reduces milk yield, impairs fertility and has implications for animal welfare. During involution, the uterus is usually exposed to multiple potential bacterial pathogens which are cleared by successful orchestration of the local inflammatory response. Unsuccessful resolution leads to the development of disease. The aim of this study was to characterize the local innate immune response in the uterus during physiological involution using histopathological and molecular analyses in 9 cows, 2 weeks after calving (early postpartum, EPP), and 4 cows, 9 weeks after calving (late postpartum, LPP). Uterine biopsies taken from each cow were classified by histopathology, and RNA was extracted for molecular analysis. Two EPP cows were classified with a mild, 5 with a moderate and 2 with a severe inflammatory response. Relative gene expression analysis was then performed using quantitative real-time PCR (qRT-PCR) and specific primers for genes encoding Toll-like receptors (TLRs), chemokines, cytokines, acute phase proteins (APPs) and antimicrobial peptides (AMPs). TLR4, transcription factor NFKB1 and the inflammatory cytokines IFNG, IL1A, IL6, IL8, IL12A were all significantly increased in EPP cows (P<0.05). Increase in HP, SAA3, TAP and DEFB5 genes was particularly marked in cows with severe inflammation. These results reveal evidence of an inflammatory uterine environment in the early postpartum period with significant induction of both AMP and APP genes. Histopathological grades in EPP cows are underpinned by quantitative changes in gene expression. Understanding the molecular mechanisms contributing to uterine immunity in the early postpartum period may identify candidate genes associated with the resolution of inflammation.
牛子宫疾病会降低产奶量，损害繁殖力，并对动物福利产生影响。在子宫复旧期间，子宫通常会接触多种潜在的细菌病原体，而通过成功协调局部炎症反应可清除这些病原体。若炎症无法成功消退，则会导致疾病的发生。本研究的目的是通过组织病理学和分子分析，对9头母牛产后2周（产后早期，EPP）和4头母牛产后9周（产后晚期，LPP）生理复旧期间子宫内的局部先天性免疫反应进行特征描述。从每头母牛采集的子宫活检组织进行组织病理学分类，并提取RNA用于分子分析。2头EPP母牛被分类为轻度炎症反应，5头为中度炎症反应，2头为重度炎症反应。然后使用定量实时PCR（qRT-PCR）和针对编码Toll样受体（TLR）、趋化因子、细胞因子、急性期蛋白（APP）和抗菌肽（AMP）的基因的特异性引物进行相对基因表达分析。EPP母牛中TLR4、转录因子NFKB1以及炎性细胞因子IFNG、IL1A、IL6、IL8、IL12A均显著增加（P<0.05）。HP、SAA3、TAP和DEFB5基因的增加在重度炎症母牛中尤为明显。这些结果揭示了产后早期子宫存在炎性环境的证据，同时AMP和APP基因均有显著诱导。EPP母牛的组织病理学分级以基因表达的定量变化为基础。了解产后早期子宫免疫的分子机制可能会识别出与炎症消退相关的候选基因。
