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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

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29 nov. 2024
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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, **