• Journal of Society of Preventive Korean Medicine
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• v.4 no.2
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• pp.294-312
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• 2000

The effects of Gamisoohwabunchungum and Gamisoohwabunchungum plus Cervi Pantotrichum Cornu on rats with nephrosis induced by a single tail-intravenous injection of PAN(puromycin amjnonucleoside), 2.5mg/100g of body weight was evaluated in the present study. The effects of Gamisoohwabunchungum and Gamisoohwabunchungum plus Cervi Pantotrichum Cornu on PAN nephrosis was evaluated by measuring the concentrations of albumin, total protein, total lipid, cholesterol, triglyceride in the serum and the amount of protein in the urin. The results are summarized as follows; 1 In The Gamisoohwabunchungum and Gamisoohwabunchungum plus Cervi Pantotrichum Cornu group as compared to The Control group, The concentrations of albumin and total protein in the serum were significantly increased. 2. In The Gamisoohwabunchungum and Gamisoohwabunchungum plus Cervi Pantotrichum Cornu group as compared to The Control group, The concentrations of total lipid, cholesterol and triglyceride in the serum were significantly decreased 3. In The Gamisoohwabunchungum and Gamisoohwabunchungum plus Cervi Pantotrichum Cornu group as compared to The Control group, The amount of urine protein during 24 hours were significantly decreased . 4. In The Gamisoohwabunchungum plus Cervi Pantotrichum Cornu group as compared to The Gamisoohwabunchungum group, The concentrations of total protein in the serum were significantly increased. 5. In The Gamisoohwabunchungum plus Cervi Pantotrichum Cornu group as compared to The Gamisoohwabunchungum group, The concentrations of cholesterol and triglyceride in the serum were significantly decreased To conclude, it can be inferred that Gamisoohwabunchungum and Gamisoohwabunchungum plus Cervi Pantotrichum Cornu have the effects of improving proteinuria, hypoproteinemia, hyperlipidemia in nephrotic syndrome.

• The Korean Journal of Physiology and Pharmacology
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• v.13 no.1
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• pp.1-7
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• 2009

Sodium retention is a hallmark of nephrotic syndrome. We investigated whether sodium retention is associated with changes of natriuretic peptide system at different stages (i.e., a sodium retaining stage and a compensatory stage) of nephrotic syndrome. At day 7 after PAN(puromycin aminonucleoside) injection, the urinary excretion of sodium was decreased, along with the development of ascites and positive sodium balance. The plasma and urinary ANP(atrial natriuretic peptide) immunoreactivities were increased. ANP mRNA expression was increased in the heart and kidney, whereas that of NPR(natriuretic peptide receptor)-A and NPR-C mRNA was decreased in the kidney. The expression of NEP was decreased in the kidney. At day 14, urinary excretion of sodium did not differ from the control. The plasma ANP level and heart ANP mRNA expression returned to their control values. The expression of ANP mRNA in the kidney was increased in association with increased urinary ANP immunoreactivities. The expression of NPR-A in the kidney became normal, whereas that of NPR-C kept decreased. The expression of NEP(neutral endopeptidase) remained decreased. These findings suggest that the increased renal ANP synthesis in association with decreased metabolism via NEP and NPR-C may play a compensatory role against the development of sodium retention in nephrotic syndrome. The decreased of NPR-A expression in the kidney may contribute to the ANP resistance at day 7. The subsequent recovery of NPR-A expression may play a role in promoting sodium excretion in later stage(at day 14).

• Biomedical Science Letters
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• v.5 no.1
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• pp.75-84
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• 1999

In order to study the effects of $\alpha$-tocopherol acetate in glomerular injury, the minimal change nephrosis disease was induced by puromycin aminonucleoside (PAN) in spontaneously hypertensive rats, and we examined biochemical analysis in serum and morphological changes. The experimental animals were divided to control, PAN-treated (30 mg/kg, I.p.), vitamin E-treated (200 mg/kg, P.O.), and PAN+vitamin E-treated groups. After PAN injection, the rate of increase of body weight was lower than the other treatments. In addition, at 8 days after PAN injection, total protein content in serum was the lowest, whereas both blood urea nitrogen and serum creatinine contents were the highest in all experimental groups, which their changes of serum parameters were statistically significant. In morphological changes, the glomerular tissue at 8 days after PAN injection clearly showed obstruction of urinary space and proliferation of mesangial cells, and that loss and fusion of pedicles, vacuolization and edema of endothelial cells, and thickness of basal lamina were ultrastructurally showed in the glomerulus. Glomerular injury was significantly prevented by administration of vitamin E having an antioxidant effect. It suggested that the glomerular injury induced by PAN was accelerated by hypertension, and renal dysfunction might be induced by oxidative injury.

• Childhood Kidney Diseases
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• v.15 no.2
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• pp.138-145
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• 2011

Purpose : To test whether the expression of ${\beta}$-catenin, a component of podocyte as a filtration molecule, would be altered by puromycin aminonucleoside (PAN) in the cultured podocyte in vitro. Methods : We cultured rat glomerular epithelial cells (GEpC) with various concentrations of PAN and examined the distribution of ${\beta}$-catenin by confocal microscope and measured the change of ${\beta}$-catenin expression by Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR). Results :We found that ${\beta}$-catenin relocalized from peripheral cytoplasm to inner cytoplasm, therefore, intercellular separations were seen in confluently cultured cells by high concentrations of PAN in immunofluorescence views. In Western blotting of GEpC, PAN ($50{\mu}g/mL$) decreased ${\beta}$-catenin expression by 34.9% at 24 hrs and 34.3% at 48 hrs, compared to those in without PAN condition (P<0.05). In RT-PCR, high concentrations ($50{\mu}g/mL$) of PAN also decreased ${\beta}$-catenin mRNA expression similar to protein suppression by 25.4% at 24 hrs and 51.8% at 48 hrs (P<0.05). Conclusion : Exposure of podocytes to PAN in vitro relocates ${\beta}$-catenin internally and reduces ${\beta}$-catenin mRNA and protein expression, which could explain the development of proteinuria in experimental PAN-induced nephropathy.

• Childhood Kidney Diseases
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• v.3 no.1
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• pp.35-41
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• 1999

Purpose The single administration of PAN(Puromycin-Aminonudeoside) to rats results in nephropathy that are similar to human minimal change nephrotic syndrome. Recently several studies indicate the pathophyslological importance of oxygen free radicals in rats with PAN-induced nephrosis. This study was conducted to evaluate the effect of $\alpha$-tocopherol, an oxygen free radical scavenger, on the histologic and biochemical changes of PAN-induced nephrosis in rats. Methods : Twenty-one Sprague-Dawley rats weighing 180-300 gm were divided into 3 groups. In group I (control group), the rats were given saline intraperitoneally for 12 days, in group II the rats were given PAN 7.5mg/100g of body weight intravenously one time and group III PAN intravenously, followed by $\alpha$-tocopherol 0.5 mg/100g of body weight jntramuscularly for 12 days. Twenty four hour urinary protein and creatinine excretion were measured on day 0, 5, 11 and 18. On the 18th day, rats were sacrificed for the determination of total serum protein, albumin and cholesterol levels. To estimate renal injuries by oxygen free radical, lipid peroxide concentration and reduced glutathione were measured in renal cortex. Histological examination in rat glomerular lesions were performed. Results : From the 5th days of PAN administration, urine protein/creatinine of group II and III were significantly increased compared the group I (P<0.05). But, urine protein/creatinine of group III was significantly lower than group II at 18th days (P<0.05). Total serum protein and albumin of group II were significantly lower than those of group III (P<0.05). Serum cholesterol of group II was significantly higher than that of group III (P<0.05). Lipid peroxide and reduced glutathione in renal cortex of group II were significantly higher than that of group I and III (P<0.05). Electron microscopic strudies of group II showed the loss of epithelial foot processes, but in group III showed preservation of epithelial foot processes. Conclusion : PAN-induced nephropathy was ameliorated significant recovery of foot process change and reduction of the urinary protein excretion by antioxidant, $\alpha$-tocopherol.

• The Journal of Korean Medicine
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• v.18 no.2
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• pp.283-315
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• 1997

The effects of Geumguesingitang and Dohongsamultang on rats with nephrosis induced by a single tail-intravenous injection of puromycin aminonucleoside(PAN), 2.5mg/l00g of body weight were evaluated in the present study. The effects of Geumguesingitang and Dohongsamultang on PAN nephrosis were evaluated by measuring (1)the concentrations of albumin, total protein, total lipid, cholesterol, triglyceride, creatinine, blood urea nitrogen(BUN) and uric acid in the serum, (2)the concentrations of protein, creatinine, glucose, occult blood and volume of the 24 hours urine and (3)the volume of intake water. The results are summerized as follows; 1. In the Control group as compared to the Normal. the amount of protein of 24 hours urine was significantly increased, the concentrations of albumin and total protein were significantly decreased. Total lipid, cholesterol and triglyceride in the serum were significantly increased. The concentrations of creatinine, BUN, uric acid in the serum, the amount of glucose and occult blood of 24 hours urine were also increased significantly. 2. In the Geumguesingitang group as compared to the Control, the increase in the amount of urinary protein during 24 hours induced by PAN was significantly suppressed, and the concentrations of total protein and albumin in the serum were significantly increased. The concentrations of total lipid, cholesterol and triglyceride in the serum were significantly inhibited. The decrease of the concentrations of creatinine and uric acid in the serum were also observed significantly. 3. In the Dohongsamultang group as compared to the Control, the increase of the amount of protein and glucose of the 24 hours urine induced by PAN were significantly inhibited, and the concentrations of total protein and albumin in the serum were increased significantly. The concentrations of total lipid, cholesterol and triglyceride in the serum were decreased significantly. The decrease of the concentrations of creatinine and uric acid in the serum were observed significantly. It can be inferred that Geumguesingitang has effects on improving proteinuria, hypoproteinemia effectively. It has an effect on hyperlipidemia significantly relieved. And relieving azotemia when nephrotic syndrome is accompanied by the acute renal failure. It can be inferred that Dohongsamultang improves hyperlipidemia effectively. It has effects on proteinuria, hypoproteinemia in nephrotic syndrome significantly relieved. And relieving azotemia when nephrotic syndrome is accompanied by the acute renal failure.

• BMB Reports
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• v.30 no.1
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• pp.55-59
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• 1997

The heat shock response is a universal stress response observed in all organisms and cultured cells. The response is regulated at both the transcriptional and translational level. Heat shocked Drosophila melanogaster Kc cells are used as the system for the study of translational regulation. In this system non-heat shock messages are associated with polysome but are not translated in a heat shocked condition. To figure out the change in the translation machinery. the effects of translation elongation inhibitors were tested on Kc cells. The result showed that the sensitivity of translation to these drugs changed in heat shocked cells. The significant changes were the decreased inhibition of heat shock protein synthesis by cycloheximide, emetine. and puromycin. and the increased inhibition of heat shock protein synthesis by verrucarin A. implying that the translation elongation mechanism in heat shocked cells changed.

• Korean Journal of Microbiology
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• v.38 no.2
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• pp.86-95
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• 2002

As a result of genome projects, the research to elucidate the function of a protein of interest has recently been well-recognized. In order to facilitate functional genomics, a useful mammalian gene expression vector is required. Using an infectious CDNA clone of BVDV pNADLclns-, we have developed a mammalian gene expression vector. In this study, a replication-competent full-length infectious CDNA clone containing puremycin acetyltransferase (pac) gene (pNADLclns-/pac) was successfully generated. The viral RNA replication and viral protein NS3 synthesis were examined by detecting metabollically $^{32}P$-labelled genomic viral RNA and immunoblotting with a mouse anti-NS3 antibody. To generate viral replicon as an expression vector, we examine if the viral structural genes (C, E0, El, E2) are required for viral replication by deletion analysis. As a result, all of the structural proteins are dispensable for viral replication per se, but essential for infectious viral particle formation. Based on our deletion analysis, we have generated a replication-competent BVDV viral replicon (pNADLclns-/pac/${\Delta}S$), whose structural genes are all deleted. In addition to NADLclns- /pac/${\Delta}S$, NADLclns-/ luc/${\Delta}S$ viral replicon containing luciferase gene as a reporter was constructed and fecund to be replication-compotent in HeLa and BHK cells as well as MDBK cells. Therefore, BVDV viral replicon developed in our study will be a useful tool to express a protein of interest in various mammalian cells.

• KSBB Journal
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• v.13 no.3
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• pp.295-302
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• 1998

Human Erythropoietin (EPO) gene is cloned in quail fibrosarcoma cell, QT35. Because molecular weight of EPO is similar to that of serum albumin, cell culture with serum containing medium makes purification of EPO very difficult. Using fractional factorial study, we have developed serum free medium for the recombinant QT35 cell lines, QT N4D4 and QT SY-IMP, which have cloned EPO with glutamine synthetase (GS) gene amplification system and with puromycin selective marker, respectively. Among the seven frequently used medium components, fibronectin, BSA, and EGF were the most important for EPO production. However, sufficient fibronectin supplement to the medium did not make any good attachment of QT35 to culture plate over 3 days. Therefore, to maximize EPO production, we attempted a medium-shift at confluence from serum containing medium to serum free medium(QT SFM6). Using the medium-shift protocol with QT SFM6, nearly the same productivity of EPO was achieved comparing with that without medium-shift. This result was true in both QT35 cell lines in three types of culture, i.e. T flask, microcarrier and roller bottle cultures.

• BMB Reports
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• v.48 no.3
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• pp.139-146
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• 2015

Adaptive brain function and synaptic plasticity rely on dynamic regulation of local proteome. One way for the neuron to introduce new proteins to the axon terminal is to transport those from the cell body, which had long been thought as the only source of axonal proteins. Another way, which is the topic of this review, is synthesizing proteins on site by local mRNA translation. Recent evidence indicates that the axon stores a reservoir of translationally silent mRNAs and regulates their expression solely by translational control. Different stimuli to axons, such as guidance cues, growth factors, and nerve injury, promote translation of selective mRNAs, a process required for the axon's ability to respond to these cues. One of the critical questions in the field of axonal protein synthesis is how mRNA-specific local translation is regulated by extracellular cues. Here, we review current experimental techniques that can be used to answer this question. Furthermore, we discuss how new technologies can help us understand what biological processes are regulated by axonal protein synthesis in vivo.