Effect of milrinone on the inflammatory response and NF-kB activation in renal ischemia-reperfusion injury in mice.

BACKGROUND: Milrinone increases intracellular adenosine 3',5'-cyclic monophosphate concentration and enhances vascular relaxation. Nuclear factor-kappa B (NF-kB) plays a key role in inflammatory responses during ischemia-reperfusion (I/R) injury. We aimed to investigate the effect of milrinone on the inflammatory responses and NF-kB activation in renal I/R injury in mice. METHODS: Thirty C57BL/6 mice were allocated into 3 groups. In group S (n = 10), only right nephrectomy was done. In group C (n = 10), the left kidney was subjected to 30 min of ischemia after right nephrectomy. In group M (n = 10), milrinone (5 µg/kg) was administered before ischemia. After 24 hours of reperfusion, the serum creatinine was measured, kidney samples were obtained for histology, and expressions of NF-kB and proinflammatory cytokines were analyzed. RESULTS: In group C, the serum creatinine concentration was markedly elevated, compared with group S. Creatinine concentration in group M was also elevated, but it was significantly lower than that in group C. Histologic evidence of renal damage was severe in group C, but it was improved in group M. In groups C and M, expression of NF-kB, tumor necrosis factor-α (TNF-α), intercellular adhesion molecule-1 (ICAM-1), monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-2 (MIP-2) mRNA increased significantly compared with group S (P < 0.05). But group M showed a lower expression of NF-kB, TNF-α, ICAM-1, MCP-1 and MIP-2 mRNA than group C (P < 0.05). CONCLUSIONS: Milrinone treatment attenuates the renal inflammatory response and activation of NF-kB, resulting in improvement of renal function and tissue injury.

Helicobacter pylori Infection Enhances Gastric Mucosal Inflammation in Individuals Carrying the 260-T Allele of the CD14 Gene.

BACKGROUND/AIMS: We aim to evaluate the association between promoter polymorphism of the clusters of differentiation 14 (CD14) gene and Helicobacter pylori-induced gastric mucosal inflammation in a healthy Korean population. METHODS: The study population consisted of 267 healthy subjects who visited our hospital for free nationwide gastric cancer screening. Promoter polymorphism at -260 C/T of the CD14 gene was determined by polymerase chain reaction and restriction fragment length polymorphism analysis. The severity of gastric mucosal inflammation was estimated by a gastritis score based on the sum of the values of the grade and activity of the gastritis. Expression of soluble CD14 (sCD14) was assessed by quantitative sandwich ELISA. RESULTS: CD14 polymorphism was not associated with H. pylori infection. There were no significant differences in gastritis scores among the genotype subgroups, but subjects carrying the CD14 -260 CT/TT genotype had significantly higher sCD14 levels than those carrying the CC genotype. Subjects with the 260-T allele of the CD14 gene and H. pylori infection had significantly higher sCD14 levels than those with the same genotype but without infection. CONCLUSIONS: In individuals with the T allele at the -260 site of the promoter region of the CD14 gene, H. pylori infection accentuates gastric mucosal inflammation.

Association between genetic polymorphisms of NOD 1 and Helicobacter pylori-induced gastric mucosal inflammation in healthy Korean population.

BACKGROUND: Gastric cancer is supposed to be a result of inflammation induced by Helicobacter pylori (H. pylori) infection. Nucleotide-binding oligomerization domain 1 (NOD 1) is required for the innate immune response to H. pylori. We aim to investigate whether single nucleotide polymorphism (SNP) in NOD 1 gene is associated with H. pylori-induced gastric mucosal inflammation in a healthy Korean population. METHODS: The study was conducted on 412 adults who visited two different healthcare centers for health examinations. The G796A (E266K) NOD 1 SNP was detected by using polymerase chain reaction/restriction fragment length polymorphism. A gastritis score was calculated by the summed values of the grade and the activity of gastritis scored according to the updated Sydney system. The expression of IL-8 and COX-2 mRNA was assessed by quantitative reverse transcription polymerase chain reaction. In the group with H. pylori infection, the complete screening of the genes comprising the cag PAI was performed. RESULTS: The genotype frequencies were 26.7% (AA type), 58.3% (GA), and 15.0% (GG). In H. pylori-positive patients, gastritis score of the AA genotype was significantly higher than those of the others (p = .04). Also, the IL-8 and COX-2 mRNA levels increased in the AA genotype. In the group with H. pylori infection, 31.9% were found to carry the complete cag PAI. When the subjects were infected with intact cag PAI, the IL-8 and COX-2 mRNA levels were significantly high in AA genotype. CONCLUSION: G796A (E266K) NOD 1 polymorphism is closely correlated with H. pylori-associated gastric mucosal inflammation in the Korean population.

Stress-responsive hypothalamic-nucleus accumbens regulation may vary depending on stressors.

This study was conducted to determine if the stress-responsive hypothalamic-nucleus accumbens (NAc) regulation is a stressor specific event. Male SD rats were subjected to restraint or cold stress for 2 h, and then mRNA expression of corticotropin-releasing hormone (CRH) in the hypothalamic paraventricular nucleus (PVN) was examined by in situ hybridization and the plasma corticosterone levels by radioimmunoassay. Neuronal activations in the PVN and the NAc were examined by c-Fos immunohistochemistry and the brain GABA contents by HPLC. Both restraint and cold stresses increased c-Fos expression in the PVN and the plasma corticosterone; however, CRH expression in PVN was increased only by restraint, but not by cold, stress. Restraint stress significantly increased the NAc neuronal activation, but cold stress failed to do so. Restraint stress increased the NAc-GABA contents and cold stress did the hypothalamic GABA. Results suggest that the HPA axis regulation responding to restraint stress, but not cold stress, may involve the NAc neuronal activation in relation with GABAergic neurotransmission. Additionally, CRH expression in the PVN may not play a major role in the elevation of plasma corticosterone responding to cold stress.

Association between Genetic Instability and Helicobacter pylori Infection in Gastric Epithelial Dysplasia.

Background. In gastric carcinogenesis, changes of DNA methylation appear to be an early molecular event, and the genome-wide methylation state is closely correlated with the level of long interspersed nucleotide element-1 (LINE-1) methylation. In this study, we measured LINE-1 methylation level according to genetic instability and evaluated the effect of Helicobacter pylori infection on genetic instability in gastric epithelial dysplasia. Methods. Total 100 tissue samples of gastric epithelial dysplasia were analyzed. Seven loci that linked to tumor suppressor genes were used to identify significant structural chromosomal aberrations. Microsatellite status was investigated for two different microsatellite marker loci (BAT25 and BAT26). Also, we measured LINE-1 methylation level by combined bisulfite restriction analysis (COBRA-LINE-1) method. Results. There were no significant differences of LINE-1 methylation level according to chromosomal/microsatellite instability and H. pylori state. In the dysplastic lesions with H. pylori infection, LINE-1 methylation level of MSI lesion was significantly lower than that of microsatellite stable (MSS) lesion (40.23 ± 4.47 versus 43.90 ± 4.81%, P < 0.01). Conclusions. In gastric epithelial dysplasia with H. pylori infection, MSI is correlated with reduced LINE-1 methylation level. Coexistence of H. pylori infection and MSI might be a driving force of gastric carcinogenesis.

Leptin blocks the fasting-induced increase of pERK1/2 in the paraventricular nucleus of rats.

This study was conducted to define molecular mechanisms by which food deprivation increases phosphorylated extracellular signal-regulated protein kinase (pERK1/2) in the hypothalamic paraventricular nucleus of rats. pERK1/2 immunoreactivity (-ir) is markedly increased in the paraventricular nucleus by 48h of food deprivation. Treatment with RU486, glucocorticoid antagonists, during food deprivation did not affect the fasting-induced increase of pERK1/2-ir in the paraventricular nucleus, but intracerebroventricular (icv) leptin blocked the increase of pERK1/2-ir by food deprivation. Fasting-induced increases of neuropeptide Y (NPY) expression both in the arcuate nucleus and the paraventricular nucleus were also blunted by icv leptin; however, the icv NPY to satiated rats did not increase pERK1/2 in the paraventricular nucleus. These results suggest that the fasting-induced increase of pERK1/2 in the paraventricular nucleus may not be mediated either by plasma corticosterone or the hypothalamic NPY, but require leptin dis-inhibition.