Singh Laishram Kipjen, Patra Manas Kumar, Mishra Girish Kumar, Singh Vidya, Upmanyu Vikramaditya, Saxena Abhishek C, Singh Sanjay Kumar, Das Goutam Kumar, Kumar Harendra, Krishnaswamy Narayanan
Animal Reproduction Division, ICAR - Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, 243 122, India.
Animal Reproduction Division, ICAR - Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, 243 122, India.
Vet Immunol Immunopathol. 2018 Nov;205:65-71. doi: 10.1016/j.vetimm.2018.10.010. Epub 2018 Oct 25.
Inflammatory markers of endometrial origin are valuable in order to differentiate the pyometra from cystic endometrial hyperplasia in the bitch. In the present study, we hypothesized that histological categorization would distinguish the differential regulation of the proinflammatory genes in the endometrium of bitches with pyometra. Ovariohysterectomy was done on bitches with confirmatory diagnosis of pyometra (n = 18). Using endometrium to myometrium ratio of 0.79 as threshold, the uteri (n = 8/group) were categorized into hyperplastic pyometra (HP) and atrophic pyometra (AP). Two samples were excluded as the diagnosis was inconclusive. In parallel, endometrial tissue was collected for total RNA extraction to study the differential expression of TLR4, IL-6, IL-8, COX-2 and PGFS through real time PCR. Diestrus uterus of non-pyometra bitches (n = 6) served as control. The mean fold change (2) for the target genes was determined using β-actin as endogenous control and non-pyometra uterus as calibrator group. Except TLR4, other inflammatory genes were upregulated significantly by 1.82 to 3.74 times in the AP as compared to HP with maximum upregulation of COX-2 and PGFS. Further, correlation matrix with Spearman's rho revealed that IL-8 had strong positive correlation with COX-2 and PGFS in the AP group (P < 0.05). It is concluded that histological grading of pyometra into HP and AP revealed differential regulation of inflammatory cytokines and enzymes in the PG synthetic pathway in the canine endometrium that has diagnostic potential under clinical settings.
子宫内膜来源的炎症标志物对于区分母犬子宫蓄脓和囊性子宫内膜增生很有价值。在本研究中,我们假设组织学分类能够区分子宫蓄脓母犬子宫内膜中促炎基因的差异调节。对确诊为子宫蓄脓的母犬(n = 18)进行了卵巢子宫切除术。以子宫内膜与子宫肌层的比例0.79为阈值,将子宫(n = 8/组)分为增生性子宫蓄脓(HP)和萎缩性子宫蓄脓(AP)。由于诊断不明确,排除了两个样本。同时,收集子宫内膜组织用于提取总RNA,通过实时PCR研究TLR4、IL-6、IL-8、COX-2和PGFS的差异表达。非子宫蓄脓母犬的间情期子宫(n = 6)作为对照。以β-肌动蛋白作为内参,非子宫蓄脓子宫作为校准组,测定目标基因的平均倍数变化(2)。与HP相比,AP中除TLR4外,其他炎症基因显著上调1.82至3.74倍,COX-2和PGFS上调最多。此外,Spearman相关系数的相关矩阵显示,AP组中IL-8与COX-2和PGFS呈强正相关(P < 0.05)。结论是,将子宫蓄脓组织学分级为HP和AP显示,犬子宫内膜中PG合成途径中的炎症细胞因子和酶存在差异调节,这在临床环境下具有诊断潜力。