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The regulatory action of acetylcholine and its receptors on B4 and C4 leukotriene formation in the porcine endometrium after experimental inflammogenic Escherichia coli infection

Barbara Jana

Barbara Jana

Division of Reproductive Biology, Institute of Animal Reproduction and Food Research of the Polish Academy of SciencesOlsztyn, Poland
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Jana, Barbara
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Jarosław Całka

Jarosław Całka

Department of Clinical Physiology, Faculty of Veterinary Medicine, University of Warmia and MazuryOlsztyn, Poland
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Całka, Jarosław
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Michał Bulc

Michał Bulc

Department of Clinical Physiology, Faculty of Veterinary Medicine, University of Warmia and MazuryOlsztyn, Poland
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Bulc, Michał
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Dominika Kawka

Dominika Kawka

Division of Reproductive Biology, Institute of Animal Reproduction and Food Research of the Polish Academy of SciencesOlsztyn, Poland
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Kawka, Dominika
  
Nov 29, 2024
The regulatory action of acetylcholine and its receptors on B4 and C4 leukotriene formation in the porcine endometrium after experimental inflammogenic Escherichia coli infection's Cover Image
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Published Online: Nov 29, 2024
Page range: 571 - 581
Received: Jun 21, 2024
Accepted: Nov 22, 2024
DOI: https://doi.org/10.2478/jvetres-2024-0066
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© 2024 Barbara Jana et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1.

Western blot results for the effects of acetylcholine (ACh) alone or with antagonists of muscarinic 2 and 3 receptor subtypes (M2R/M3R) and α-7 nicotinic ACh receptor (α-7 nAChR) on 5-lipoxygenase (5-LO; A), leukotriene (LT) A4 hydrolase (LTAH; B) and LTC4 synthase (LTCS; C) protein abundances in the endometrium of gilts from the control (grey bars, n = 5) and E. coli (black bars, n = 5) groups. Blots with representative bands for each enzyme in particular groups are depicted in Supplementary Figure 2. a, b, c – different letters for any pair of values within a group indicate significant difference (P-value < 0.05–0.001) between the control values (CV), action of ACh and actions of particular antagonists with ACh; *** – statistical difference between groups for the same type of treatment (P < 0.001)
Western blot results for the effects of acetylcholine (ACh) alone or with antagonists of muscarinic 2 and 3 receptor subtypes (M2R/M3R) and α-7 nicotinic ACh receptor (α-7 nAChR) on 5-lipoxygenase (5-LO; A), leukotriene (LT) A4 hydrolase (LTAH; B) and LTC4 synthase (LTCS; C) protein abundances in the endometrium of gilts from the control (grey bars, n = 5) and E. coli (black bars, n = 5) groups. Blots with representative bands for each enzyme in particular groups are depicted in Supplementary Figure 2. a, b, c – different letters for any pair of values within a group indicate significant difference (P-value < 0.05–0.001) between the control values (CV), action of ACh and actions of particular antagonists with ACh; *** – statistical difference between groups for the same type of treatment (P < 0.001)

Fig. 2.

ELISA results for the effects of acetylcholine (ACh) alone or with antagonists of muscarinic 2 and 3 receptor subtypes (M2R/M3R) and α-7 nicotinic ACh receptor (α-7 nAChR) on the leukotriene (LT)B4 (A) and LTC4 (B) secretion from the endometrium of gilts from the CON (grey bars, n = 5) and E. coli (black bars, n = 5) groups. a, b, c – different letters for any pair of values within a group indicate significant difference (P-value < 0.05–0.001) between the control values (CV), action of ACh and actions of particular antagonists with ACh; * P < 0.05, ** P < 0.01, *** P < 0.001 – statistical difference between groups for the same type treatment
ELISA results for the effects of acetylcholine (ACh) alone or with antagonists of muscarinic 2 and 3 receptor subtypes (M2R/M3R) and α-7 nicotinic ACh receptor (α-7 nAChR) on the leukotriene (LT)B4 (A) and LTC4 (B) secretion from the endometrium of gilts from the CON (grey bars, n = 5) and E. coli (black bars, n = 5) groups. a, b, c – different letters for any pair of values within a group indicate significant difference (P-value < 0.05–0.001) between the control values (CV), action of ACh and actions of particular antagonists with ACh; * P < 0.05, ** P < 0.01, *** P < 0.001 – statistical difference between groups for the same type treatment

Effects of acetylcholine (ACh) alone or antagonists of muscarinic 2 and 3 receptor subtypes (M2R/M3R) and α-7 nicotinic ACh receptor (α-7 nAChR) alone on leukotriene (LT)B4 and LTC4 secretion from the endometrium of gilts determined by ELISA

Treatment Group Concentrations (ng/g tissue)
LTB4 LTC4
Control value Control 2.41 ± 0.14a 0.85 ± 0.11a
E. coli 3.37 ± 0.18a, ** 1.29 ± 0.08a, *
ACh Control 3.41 ± 017b 1.45 ± 0.1b
E. coli 4.25 ± 0.26b, * 1.82 ± 0.09b
M2R antagonist Control 2.42 ± 0.23a 0.98 ± 0.09a
E. coli 3.26 ± 0.22a, * 1.33 ± 0.11a
M3R antagonist Control 2.49 ± 0.15a 0.81 ± 0.12a
E. coli 3.09 ± 0.27a 1.27 ± 0.15a, *
α-7 nAChR antagonist Control 2.31 ± 0.17a 0.92 ± 0.07a
E. coli 2.99 ± 0.26a 1.16 ± 0.14a

Western blot results for the effects of acetylcholine (ACh) alone or antagonists of muscarinic 2 and 3 receptor subtypes (M2R/M3R) and α-7 nicotinic ACh receptor (α-7 nAChR) alone on the 5-lipoxygenase (5-LO), leukotriene (LT)A4 hydrolase (LTAH) and LTC4 synthase (LTCS) protein abundances in unchallenged and E_ coli-challenged endometria of gilts

Treatment Group Protein expression (arbitrary units)
5 LO LTAH LTCS
Control value Control 0.12 ± 0.01a 1.82 ± 0.24a 0.73 ± 0.04a
E. coli 0.76 ± 0.12a, *** 1.71 ± 0.07a 2.31 ± 0.36a, ***
ACh Control 0.65 ± 0.07b 3.06 ± 0.11b 2.15 ± 0.09b
E. coli 1.63 ± 0.21b, *** 2.81 ± 0.13b 5.16 ± 0.42b, ***
M2R antagonist Control 0.09 ± 0.01a 1.91 ± 0.16a 0.62 ± 0.06a
E. coli 0.83 ± 0.09a, *** 1.23 ± 0.09a, ** 3.29 ± 0.23a, ***
M3R antagonist Control 0.13 ± 0.05a 1.87 ± 0.08a 0.54 ± 0.07a
E. coli 0.69 ± 0.14a, ** 1.65 ± 0.11a 2.86 ± 0.31a, ***
α-7 nAChR antagonist Control 0.11 ± 0.08a 1.41 ± 0.05a 0.63 ± 0.05a
E. coli 0.68 ± 0.15a, ** 1.48 ± 0.07a 1.99 ± 0.38a, **
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