The effect of dietary supplementation of nitric oxide donor and inhibitor on nNOS expression in and motility of the small intestine of broilers

Abstract

We investigated the effect of dietary supplementation of sodium nitroprusside (SNP), a nitric oxide (NO) donor, and N-nitro-L-arginine methyl ester (L-NAME), a NO inhibitor, on neuronal nitric oxide synthase (nNOS) expression in and motility of small intestinum in broilers. A total of 560, one-day-old Ross 308 hybrid mixed sex broiler chicks were divided randomly into one control and seven treatment groups for a 42 day feeding trial including starter phase (0-21 days) and grower phase (22-42 days). The control group was fed a basal diet and the experimental groups were the fed basal diet supplemented with 25, 50, 100 and 200 mg/kg SNP and 25, 50 or 100 mg/kg L-NAME. Ten chickens from each group were sacrificed to collect samples on days 21 and 42. The expression patterns of nNOS immunoreactivity in nerve fibers were determined by immunohistochemistry. In the contractility studies, longitudinal isolated strips of duodenum, jejunum and ileum were treated with 10(-5) M L-arginine and 10(-4) M SNP. Immunohistochemistry revealed that nNOS expression was not detectable in the duodenum or ileum of either the control or experimental groups. On the other hand, nNOS immunoreactivity in the jejunum control group showed a strong reaction on day 21, but the reaction was weak on day 42. nNOS expression clearly was suppressed on day 21 by the diet supplemented with L-NAME, while the diet supplemented with SNP stimulated nNOS expression on day 21. Contractility experiments revealed that spontaneous contractility of isolated strips of duodenum, jejunum and ileum showed no significant difference among groups. Spontaneous contractions of all strips were inhibited by L-arginine and SNP in all groups. The percentage inhibition rate of spontaneous contractions of jejunum application on days 21 and 42 after L-arginine decreased in the group supplemented with 100 mg/kg L-NAME. The percentage inhibition rate on day 21 after SNP application decreased in both groups that received 50 and 100 mg/kg L-NAME. We demonstrated the expression pattern of nNOS in nerve fibers in jejunum of broiler chickens. Contractility studies revealed that the NOS-NO pathway may play a role in smooth muscle contraction of small intestine of chickens. Feeding strategies that supplement NO donor and NO inhibitor can be of physiological importance to small intestine motility owing to alteration of nNOS expression in the jejunum.

The Effect of L-Arginine on Growth Performance, Some Serum Biochemical Parameters and Duodenal Motility in Broilers

“…However, it was observed the inhibition rate did not show any significant difference between [33] . Similarly, it has already been reported that SNP, a exogenous NO donor and L-NAME, a selective NOS inhibitor changes the nNOS expression in jejunum but not in duodenum and ileum in broilers [34] . In this study, no difference was observed in the inhibition of duodenum contractility between groups, in vitro.…”

The effect of dietary L-arginine intake on the level of antibody titer, the relative organ weight and colon motility in broilers

“…The relaxation of smooth muscle of intestine system is mediated by NO, which is released from NANC nerve fibers via NOS enzyme following electrical stimulation of intestine (Toda et al 1990). Bulbul et al (2013) reported that duodenum motility might be inhibited by NOS donor in broilers. In this study, the amplitude of mid colon contractions did not differ between groups (Table 5).…”

“…Nitric oxide released from non-adrenergic non-cholinergic (NANC) nerves in intestines decreases intestinal motility in rat (Lie and Rand 1990), mice (Bani et al 2002) and dog (Bult et al 1990), in vitro. Similarly, the diet supplemented with sodium nitroprusside (SNP), an exogenous NO donor increases the nNOS expression in jejunum of broilers, while N-nitro-L-arginine methyl ester (L-NAME), a NOS inhibitor, decreases the nNOS expression (Bulbul et al 2013).…”

17β-Estradiol Inhibites Nitric Oxide-cGMP-Dependent Pathway but may Activate Independent Pathway in Small Intestinum of Ovariectomized Rat

“…Therefore, a resting tension of 2 g was applied to the tissues. Electrical field stimulation was then used to define submaximal contraction, carried out at various frequencies (2,4,8,16, 32 and 64 Hz) and voltage (10, 20, 30 and 40 voltage). In order to determine endogenous NO activity, L-arginine (10 -5 M) (Sigma, Cat # A5131) was added to the Krebs' solution.…”

“…Firstly, NO 2 reduces levels of Ca ++ within a cell and increases the permeability of K + channels. Thus, it hyperpolarizes the plasma membrane [15,16] . Secondly, cGMP blocks miyozin/ aktin interaction by activating cGMP dependent protein kinaz (PKG) which leads to the dephosphorization of light miyozin chains and which is reported to play a key role in NO/cGMP signals [15] .…”