The Regulation of ERK, GSK3beta and AKT after Acute Ethanol Exposure and Withdrawal in SH-SY5Y Human Neuroblastoma Cell-line / 신경정신의학

OBJECTIVES: This study aimed to demonstrate the activities and phosphorylation changes induced by acute ethanol treatment and withdrawal conditions in the intracellular signal transduction molecules [such as extracellular signal-regulated kinase (ERK), glycogen synthase kinase 3beta (GSK3beta), and Akt] of the SH-SY5Y neuroblastoma cell line. METHODS: The acute treatment exposed SH-SY5Y cells to 100 mM ethanol, and we took samples 30 minutes, 60 minutes, and 24 hours after initiating this treatment. After 24 hours' continuous ethanol treatment, we initiated ethanol withdrawal, taking samples at 30 minutes and 60 minutes. We assayed the kinase phosphorylations via an immunoblot analysis using phosphorspecific antibodies, quantified by optical densitometry. RESULTS: Ethanol treatment induced a transient increase in phosphorylation of GSK3beta and Akt at 30 minutes but failed to change the phosphorylation level of ERK. Ethanol withdrawal induced a transient ERK phosphorylation increase at 30 minutes, but it had no effect on the phosphorylation of GSK3beta or Akt. CONCLUSION: The results indicate that the ethanol-induced cellular response includes the ERK, GSK3beta, and Akt systems. In particular, the ERK pathway may play a role in the acute withdrawal response. This also suggests that a relatively short exposure to ethanol, such as the 24-hour exposure in this study, can induce functional adaptation within a cell.

The development of an efficient rat hepatic cirrhosis model

INTRODUCTION: Applying clinical conditions to on experimental animals forto verifverifyingy the mechanism of disease and drug effects is crucial. Cirrhotic livers induced by Hepatitis B virus are frequent, and eEspecially in Korea where a great deal of more liver-related diseases occurs, cirrhotic livers induced by Hepatitis B virus are frequent, and, such viral-induced cirrhosis, and this often impedes other medical treatments. Therefore, creating a proper elucidating properly deriveding cirrhosis method in animal model to simulate the actual pathophysiology of cirrhosis can benefit future researches. AIMS: We wanted toTe testing various hypotheticalsized methods of inducing cirrhosis in animal models, and we wanted the model to have a with higher rate of reproducibility. METHOD: To induce cirrhotic liver, thioacetamide (Sigma, St. Louis, USA) wasis given either freely via oral intaken or it wasand injected into the peritoneal space ofn Sprague-Dawley(SD) rats. The SD rats wereare divided into four groups: the oOral intake gGroup 1 ((N=10, 0.03%, 13 weeks), the oOral intake gGroup 2 (N=20, 0.04%, 30 weeks), the iIntraperitoneal Injected gGroup 1 (N=10, 300mg/kg, 12 weeks (3 times per week for first 2 weeks, 2 times per week for next 10 weeks) and the iIntraperitoneal Injected gGroup 2 (N=20, 300mg/kg, 2 times per week for 16 weeks). The mMortality rate of the tested subjects is recorded, and a visual test of the livers is performed at the end of the experiment, a visual test of the livers is performed. Also, the extracted liver cells that were dyed with Trichrome are compared to evaluate the extent of the liver cirrhosis. RESULT: For theIn oral intake group 1, no loss of occurred until wWeek 13, and 5 of the SD rats (50%) showed signs of liver cirrhosis by the Trichrome dye test. However, the extent of cirrhosis greatly differed betweenfrom each of the subjects. ForIn the oral intakae group 2, no loss occurred until wWeek 30. 20 of the SD rat (100%) in this group possessed a cirrhotic liver. However, the weight of the cirrhoscirrhotic liversis differed from a minimum of 231g to a maximum of 770g. For theIn Injected Group 1, 4 tested subjects (40%) died between wWeeks 3 and 4; however, the rest of them survived and they all revealed a signs of cirrhosis. ForIn the iInjected Group 2, only 3 tested subjects (15%) died, and after wWeek 16, 17 survivors (100%) showed a signs of cirrhosis. CONCLUSION: The short-term oral administration of thioacetamide only induced a minimal amount of cirrhosis;, thus, a longer period of consumption is suggested. Injection of thioacetamide into the peritoneum resulted in higher death rate when thoacetamide wasis injected frequently. Therefore, selecting a proper method to create a cirrhotic liver, with considering the reproducibility, on cirrhotic liver, the survival rate of the experimental animals, and the length of the experiment, isare strongly suggested for creating an animal model of cirrhotic liverfor further experiments.

The Effects of air-borne particulate matters on the Alveolar Macrophages for the iNOS Expression and Nitric Oxide with Nitrotyrosilated-proteins Formation / 결핵및호흡기질환

BACKGROUND: Particulate matters (PM) when inhaled is known to induce pulmonary diseases including asthma and chronic bronchitis when inhaled. Despite the epidemiological proofevidence, the pathogenesis of PM-related pulmonary diseases is unclearremain poorly understood. METHODS: Primary alveolar macrophages were harvested from the SPF and inflammatory rats by bronchioalveolar lavage (BAL). The cultured primary alveolar macrophages were treated with the medium only, PM only (5~40 microgram/cm2), LPS (5ng/ml) only, and PM with LPS for 24 and 48 hours. The level of secreted nitric oxide (NO) was assayed from the cultured medium by using the Griess reaction. The cultured cells were utilized for the western blotting against the inducible nitric oxide synthase (iNOS) proteins. Immunocyto- chemical staining against the iNOS and NT-proteins were performed in cells that cultured in the Lab-Tek(R) chamber slide after treatments. RESULTS: The PM that utilizein this experiments induced NO formation with iNOS expression in the cultured SPF and inflammatory rats alveolar macrophages, by itself. When the cells were co-treated with PM and LPS, there was a statistically significant synergistic effect on NO formation and iNOS expression over the LPS effect. The cells from the sham control showed minimal immunoreactivity for the NT-proteins. Significantly higher quantities of NT-proteins were detected in the PM and PM with LPS co-treated cells than from the sham control. CONCLUSION: Increased iNOS expression and NO formation with increased NT-proteins formation might be involved in the pathogenesis of PM-induced lung injury.