• 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.

• 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 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.

• 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.

• Childhood Kidney Diseases
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• v.7 no.1
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• pp.9-15
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• 2003

Purpose : Several studies have suggested that hyperlipidemia might be a causative factor contributing to the progression of initial glomerular injury through the development of glomerulosclerosis. We examined the potential beneficial effect of atorvastatin - which blocks the rate limiting step of cholesterol synthesis by inhibiting HMG-CoA reductase - in PAN-induced nephrosis. Materials and Methods : Glomerulosclerosis was induced in Sprague-Dawley male rats by repeated administration of PAN. Sprague-Dawley male rats were divided into 3 groups : group I(control), group II(PAN 20 mg/kg, subcutaneous injection), group III(PAN 20 mg/kg subcutaneous injection and atorvastatin 50 mg/kg/day per oral). On the 11th week, upon sacrifice of the experimental animals, blood sampling, 24-hr urine collection and nephrectomy were performed. Results : Group III had significantly lower BUN and higher serum albumin($30.9{\pm}17.2\;vs.\;17.3{\pm}2.5\;mg/dL;\;2.3{\pm}0.1\;vs.\;2.5{\pm}0.2\;g/dL$, P<0.05) compared with group II. In the lipid profiles, group III was associated with a reduction in total cholesterol and LDL($291{\pm}173\;vs.\;167{\pm}72\;mg/dL:\;57{\pm}53\;vs.\;27{\pm}12\;mg/dL$, P>0.05) compared with group II. Atorvastatin administration lowered the glomerular sclerosing index significantly(26.2% vs. 13.3%, P<0.05). Conclusion : Puromycin-induced glomerulosclerosis could be ameliorated by the reduction of hyperlipidemia with atorvastatin. This suggests that hyperlipidemia contributes to the pathogenesis of glomerulosclerosis.

• BMB Reports
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• v.46 no.4
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• pp.230-235
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• 2013

Nephrin, a structural molecule, is also a signaling molecule after phosphorylation. Inhibition of nephrin phosphorylation is correlated with podocyte injury. The PINCH-1-ILK-${\alpha}$-parvin (PIP) complex plays a crucial role in cell adhesion and cytoskeleton formation. We hypothesized that nephrin phosphorylation influenced cytoskeleton and cell adhesion in podocytes by regulating the PIP complex. The nephrin phosphorylation, PIP complex formation, and F-actin in Wistar rats intraperitoneally injected with puromycin aminonucleoside were gradually decreased but increased with time, coinciding with the recovery from glomerular/podocyte injury and proteinuria. In cultured podocytes, PIP complex knockdown resulted in cytoskeleton reorganization and decreased cell adhesion and spreading. Nephrin and its phosphorylation were unaffected after PIP complex knockdown. Furthermore, inhibition of nephrin phosphorylation suppressed PIP complex expression, disorganized podocyte cytoskeleton, and decreased cell adhesion and spreading. These findings indicate that alterations in nephrin phosphorylation disorganize podocyte cytoskeleton and decrease cell adhesion through a PIP complex-dependent mechanism.