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The Role of NO in the Opium-Induced Bowel Dysfunction in the Mice
2020
Dabili Nasrabadi, Amin | Arab, Hossein Ali | Fatemi Ardestani, Syyed Ahmad | Hassanpor, Hossein
BACKGROUND: Opioids and nitric oxide (NO) are functionally linked in the regulation of intestinal motility. OBJECTIVES: To investigate the role of NO in the opium induced bowel dysfunction in mice. METHODS: Sixty-six male mice received incrementally doses of the following treatments in six groups for 5 consecutive days: 1) Opium (0.2, 0.3, 0.4, 0.5 and 0.6mg/30g/day), 2) N-nitro-L-arginine methyl ester (L-NAME, 5,7.5,10,15 and 20mg/kg/day), 3) L-arginine (5-20mg/kg/day), 4) Opium+L-NAME, 5) Opium+L-arginine and 6) distilled water. At the end of the treatment, the abdomen was opened; some pieces of duodenal and proximal colon were taken to determine NO synthase (NOS) expression and nitrite levels, and some isolated rings from those parts of small and large intestine were prepared and transferred to the organ bath system to study intestinal motility. RT-PCR was used to determine the NOS gene expression. To determine the small intestinal transit, 30 mice in six groups, were used for oral administration of charcoal+gum in vivo. RESULTS: Opium decreased amplitude of the duodenum and ileum contractions, but increased frequency of duodenal and mid colon contractions (P<0.05). While the gene expression of inducible, neuronal and endothelial NOS was increased in colon (P<0.05), a reduced neuronal and endothelial NOS gene expression was shown in duodenum. The charcoal+gum transit was decreased in opium-treated animals compared to the control group (19.9%). However, L-arginine increased this transit while L-NAME decreased it. CONCLUSIONS: Opium induced intestinal smooth muscle spasms, which result in the decreased intestinal movements. The alterations in NOS gene expression may be a compensation mechanism against opium-induced intestinal dysfunction.
Показать больше [+] Меньше [-]An immunohistochemical study on the presence of nitric oxide synthase isoforms (nNOS, iNOS, eNOS) in the spinal cord and nodose ganglion of rats receiving ionising gamma radiation to their liver
2020
Yılmaz, Osman | Soygüder, Zafer | Keleş, Ömer Faruk | Yaman, Turan | Yener, Zabit | Uyar, Ahmet | Çakır, Tahir
This study determined the presence of nitric oxide synthesis isoforms (nNOS, iNOS, and eNOS) in thoracic spinal cord segments and nodose ganglia of rats with gamma-irradiated livers. Male rats (n = 32) were divided into equal groups A, B, C, and D. In group A, the controls, no radiation was applied, while groups B, C, and D received 10 Gy of ionising gamma radiation. The rats of group B were euthanized at the end of the first day (d1), those of group C on the second day (d2), and those of group D on the third day (d3). The liver, spinal cord segments, and nodose ganglion tissues were dissected and fixed, and the liver sections were examined histopathologically. The other tissues were observed through a light microscope. Regeneration occurred at the end of d3 in hepatocytes which were radiation-damaged at the end of d1 and d2. On d1, some nNOS-positive staining was found in the neuronal cells of laminae I–III of the spinal cord and in neurons of the nodose ganglion, and on d3, some staining was observed in lamina X of the spinal cord, while none of note was in the nodose ganglion. Dense iNOS-positive staining was seen on d1 in the ependymal cells of the spinal cord and in the glial cells of the nodose ganglion, and on d3, there was still considerable iNOS staining in both tissues. There was clear eNOS-positive staining in the capillary endothelial cells of the spinal cord and light diffuse cytoplasmic staining in the neurons of the nodose ganglion on d1, and on d3, intense eNOS-positive staining was visible in several endothelial cells of the spinal cord, while light nuclear staining was recognised in the neurons of the nodose ganglion. The nNOS, iNOS, and eNOS isoforms are activated in the spinal cord and nodose ganglion of rats after ionising radiation insult to the liver.
Показать больше [+] Меньше [-]Effect of Achyranthes japonica Nakai extract on immunity and anti-inflammation in dogs
2020
Lee, Gun-Hwi | Hwang, Kyung-A | Kang, Ji-Houn | Choi, Kyung-Chul
Achyranthes japonica Nakai (A. japonica) is a medicinal herb found widely distributed throughout Korea. The biological activities of A. japonica are well-documented and include anti-fungal, anti-inflammatory, and immunity enhancement. The objective of the present study was to investigate the immune-related activities of A. japonica extract in dogs. The extract was acquired by ethanol extraction and purified by filtration. To examine the effect of A. japonica extract on immune cell viability, human lymphocytes, such as Jurkat T-cells and Ramos B-cells, were exposed to the extract. After treatment with the extract, the number of Ramos B-cells was increased, whereas Jurkat T-cells remained unaffected. Griess assay revealed decreased nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated mouse macrophage Raw 264.7 cells after exposure to A. japonica extract. To evaluate the in-vivo effect in dogs, feed containing A. japonica extract was provided to 8 dogs for 2 months. Blood samples were collected before, during, and after consumption of the feed. Peripheral blood mononuclear cells (PBMCs) were isolated from the blood samples and the number of T-cells and B-cells were assessed using flow cytometry with anti-dog fluorescein isothiocyanate (FITC)-conjugated CD3 and anti-dog phycoerythrin (PE)-conjugated CD21 antibodies, respectively. We observed a significant increase in the average number of B-cells in the PBMCs during ingestion of the feed containing A. japonica. In addition, enzyme-linked immunosorbent assay (ELISA) revealed a decrease in the levels of tumor necrosis factor-alpha (TNF-α), a pro-inflammatory cytokine, in 3 out of 8 dogs and increased levels of interleukin-10 (IL-10), an anti-inflammatory cytokine, in 4 out of 8 dogs. Taken together, we believe that these changes indicate that A. japonica extract is beneficial in improving the immunity of dogs by stimulating B-cells and inducing production of anti-inflammatory responses.
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