Jana Barbara, Całka Jarosław, Bulc Michał, Kawka Dominika
Division of Reproductive Biology, Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences, 10-748 Olsztyn, Poland.
Department of Clinical Physiology, Faculty of Veterinary Medicine, University of Warmia and Mazury, 10-718 Olsztyn, Poland.
J Vet Res. 2024 Nov 29;68(4):571-581. doi: 10.2478/jvetres-2024-0066. eCollection 2024 Dec.
Endometritis is a very common pathology in animals which changes endometrial leukotriene (LT) formation and muscarinic 2 and 3 receptor subtypes (M2R/M3R) and α-7 nicotinic acetylcholine (ACh) receptor (α-7 nAChR) expression patterns. With the relationship between ACh, its receptors and LT production remaining unclear, the role of M2R, M3R and α-7 nAChR in action of ACh on the 5-lipoxygenase (5-LO), LTA4 hydrolase (LTAH) and LTC4 synthase (LTCS) protein abundances in the inflamed porcine endometrium and on the tissue secretion of LTB4 and LTC4 were studied.
On day three of the oestrous cycle in gilts aged 7-8 months, 50 mL of either saline solution (control group, n = 5) or an suspension at 10 colony-forming units/mL ( group, n = 5), was injected into each uterine horn. Endometrial explants obtained eight days later, were incubated with ACh alone, antagonists of M2R, M3R and α-7 nAChR alone, or with ACh together with particular antagonists for 16 h. Enzyme abundances in endometrial tissue were estimated by Western blotting, and LT concentrations in medium by ELISA.
Severe acute endometritis developed in the group. In the endometrial explants from both groups, ACh elevated 5-LO, LTAH and LTCS protein abundances and LTB4 and LTC4 release. In the group, ACh-induced 5-LO and LTCS abundances and LTB4 release were increased versus the control group. In both groups, the M3R antagonist with ACh reduced all ACh-stimulated enzyme abundances and LT release in comparison to the abundances and release mediated by ACh alone. This effect on LTCS protein abundance and LTB4 release was also produced by the M2R antagonist with ACh in the group. Compared to the effect of ACh alone, exposure of the group endometrium to the α-7 nAChR antagonist with ACh led to a rise in LTAH and LTCS protein abundances and LTB4 and LTC4 secretion.
In the inflamed pig endometrium, ACh increased 5-LO, LTAH and LTCS protein abundances and LTB4 and LTC4 release by M3R, and LTCS protein abundance and LTB4 release also by M2R. By interaction with α-7 nAChR, ACh reduced LTAH and LTCS protein abundances and the release of these LTs. Thus, in an indirect manner, ACh can affect LT-controlled processes.
子宫内膜炎是动物中一种非常常见的病理状况，它会改变子宫内膜白三烯（LT）的形成以及毒蕈碱2型和3型受体亚型（M2R/M3R）和α-7烟碱型乙酰胆碱（ACh）受体（α-7 nAChR）的表达模式。由于ACh及其受体与LT产生之间的关系尚不清楚，因此研究了M2R、M3R和α-7 nAChR在ACh对炎症状态下猪子宫内膜中5-脂氧合酶（5-LO）、LTA4水解酶（LTAH）和LTC4合酶（LTCS）蛋白丰度以及对LTB4和LTC4组织分泌作用中的角色。
在7 - 8月龄后备母猪发情周期的第三天，向每个子宫角注射50 mL生理盐水（对照组，n = 5）或浓度为10个菌落形成单位/ mL的悬浮液（实验组，n = 5）。八天后获取子宫内膜外植体，分别单独与ACh、M2R拮抗剂、M3R拮抗剂和α-7 nAChR拮抗剂孵育，或与ACh及特定拮抗剂共同孵育16小时。通过蛋白质免疫印迹法估计子宫内膜组织中的酶丰度，并用酶联免疫吸附测定法测定培养基中的LT浓度。
实验组发生了严重的急性子宫内膜炎。在两组的子宫内膜外植体中，ACh均提高了5-LO、LTAH和LTCS蛋白丰度以及LTB4和LTC4的释放。与对照组相比，在实验组中ACh诱导的5-LO和LTCS丰度以及LTB4释放增加。在两组中，与单独由ACh介导的丰度和释放相比，M3R拮抗剂与ACh共同作用降低了所有ACh刺激的酶丰度和LT释放。在实验组中，M2R拮抗剂与ACh共同作用对LTCS蛋白丰度和LTB4释放也产生了这种影响。与单独使用ACh的效果相比，实验组子宫内膜与α-7 nAChR拮抗剂及ACh共同孵育导致LTAH和LTCS蛋白丰度以及LTB4和LTC4分泌增加。
在炎症状态的猪子宫内膜中，ACh通过M3R增加5-LO、LTAH和LTCS蛋白丰度以及LTB4和LTC4释放，并且通过M2R增加LTCS蛋白丰度和LTB4释放。通过与α-7 nAChR相互作用，ACh降低了LTAH和LTCS蛋白丰度以及这些LTs的释放。因此，ACh可以间接影响LT控制的过程。
