Ankara University, Faculty of Veterinary Medicine, Department of Obstetrics and Gynecology, Ankara 06110, Turkiye.
Hatay Mustafa Kemal University, Faculty of Veterinary Medicine, Department of Genetics, Hatay 31060, Turkiye.
Anim Reprod Sci. 2024 Dec;271:107621. doi: 10.1016/j.anireprosci.2024.107621. Epub 2024 Oct 24.
Pyometra is a common life-threatening inflammatory disease with a complex etiopathogenesis that develops during the diestrus stage and can be observed in elderly intact bitches. The present study evaluated five aquaporin (AQP1, AQP2, AQP3, AQP5, and AQP9) transcript abundances and immunolocalization in the uterine tissue, and investigated their relationship with uterine tissue and blood lipopolysaccharide (LPS) concentration, superoxide dismutase (SOD) and glutathione peroxidase (GPX) activity, and nitric oxide (NO) production in dogs suffering from pyometra. The study sampled 36 client-owned intact bitches from different breeds, of which 24 cases were diagnosed with pyometra. Twelve of these bitches in the diestrus stage that presented for elective ovariohysterectomy were used as the control group. Blood samples were collected into tubes without anticoagulant for serum progesterone, LPS concentration, and antioxidant activities at the time of diagnosis. Bacteriological and tissue samples from the uteri were collected after the ovariohysterectomy. The tissue samples were used to determine antioxidant activity, and hormone and toxin concentrations. Transcript abundance of uterine AQPs were determined by qPCR, and their presence and localization were determined by by immunohistochemistry. For all pyometra samples, the bacteria isolated from the uterine swabs were Escherichia coli. Compared to the control group, AQP1, AQP2, and AQP5 were downregulated more than 2-fold, whereas AQP9 was upregulated nearly 3-fold and AQP3 was upregulated more than 4-fold in the pyometra affected uteri (P<0.05). Uterine AQP1 was moderately negatively correlated with serum LPS concentration (r=-0.568, P<0.01) and tissue NO production (r=-0.407, P<0.05). AQP5 was positively correlated with serum SOD activity (r=0.485, P<0.05) and negatively correlated with serum LPS concentration (r=-0.512, P<0.05). AQP9 was negatively correlated with tissue SOD and serum GPx activity. This is the first study to identify AQP9 transcript abundance and immunolocalization in canine uterine tissue. Uterine AQP1, AQP2, AQP3, AQP5, and AQP9 transcript abundances were altered in spontaneously developed canine pyometra while AQP transcript abundance was negatively related to serum toxin concentration, NO production, and antioxidant enzyme activity. Further studies should be conducted to determine the role of altered abundances of AQPs transcripts in pyometra pathogenesis.
脓性子宫蓄脓是一种常见的危及生命的炎症性疾病,其发病机制复杂,发生于发情后期,可发生于老年未绝育犬。本研究评估了五种水通道蛋白(AQP1、AQP2、AQP3、AQP5 和 AQP9)在子宫组织中的转录丰度和免疫定位,并研究了它们与子宫组织和血液中脂多糖(LPS)浓度、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GPX)活性以及一氧化氮(NO)产生的关系患有脓性子宫蓄脓的犬。该研究从不同品种的 36 只患犬中采集样本,其中 24 例被诊断为脓性子宫蓄脓。其中 12 只为发情期接受选择性卵巢子宫切除术的犬作为对照组。在诊断时,采集无抗凝剂管中的血液样本用于检测血清孕酮、LPS 浓度和抗氧化活性。在卵巢子宫切除术时采集子宫的细菌和组织样本。组织样本用于测定抗氧化活性、激素和毒素浓度。通过 qPCR 测定子宫 AQP 的转录丰度,并通过免疫组织化学测定其存在和定位。对于所有脓性子宫蓄脓样本,从子宫拭子中分离的细菌均为大肠杆菌。与对照组相比,AQP1、AQP2 和 AQP5 的表达下调超过 2 倍,而 AQP9 的表达上调近 3 倍,AQP3 的表达上调超过 4 倍(P<0.05)。子宫 AQP1 与血清 LPS 浓度呈中度负相关(r=-0.568,P<0.01)和组织 NO 产生呈负相关(r=-0.407,P<0.05)。AQP5 与血清 SOD 活性呈正相关(r=0.485,P<0.05),与血清 LPS 浓度呈负相关(r=-0.512,P<0.05)。AQP9 与组织 SOD 和血清 GPx 活性呈负相关。这是首次在犬子宫组织中鉴定 AQP9 转录丰度和免疫定位。自发性发生的犬脓性子宫蓄脓时,子宫 AQP1、AQP2、AQP3、AQP5 和 AQP9 的转录丰度发生改变,而 AQP 转录丰度与血清毒素浓度、NO 产生和抗氧化酶活性呈负相关。应进一步开展研究以确定 AQP 转录本丰度改变在脓性子宫蓄脓发病机制中的作用。