RESUMEN
It is well known that the incidence of urinary stones is higher in men than women. Although it is believed that the lower incidence of urinary stones in women is due to a protective effect of estrogen, the mechanisms remain unclear. To clarify the relation between female sex hormones and stone matrix protein, we examined the interaction of estrogen receptor-α (ERα), estrogen receptor-related receptor-α (ERRα), and stone matrix protein osteopontin (OPN) in a rat hyperoxaluric model and in primary cultured rat kidney cells. Adult female Wistar rats were divided into 6 groups. Groups 1 and 4 consisted of normal females, Groups 2 and 5 consisted of ovariectomized females, and Groups 3 and 6 consisted of ovariectomized females receiving female sex hormone supplements. Groups 1-3 were administered distilled water, while groups 4-6 were administered 0.5% ethyleneglycol (EG). Moreover, rat kidney primary cultured cells were examined after treatment with female sex hormones under various conditions. The expressions of ERα, ERRα and OPN-mRNA in whole kidney and primary cultured cells were examined using Real-Time PCR. The expressions of OPN and ERRα-mRNA were suppressed by ovariectomy. Supplementation with female sex hormones increased the expression of OPN and ERRα-mRNA. In contrast, the expression of ERα-mRNA was increased by ovariectomy and suppressed by supplementation with female sex hormones. The results of the mRNA expression in primary cultured cells matched those in the hyperoxaluric model rats. Although the reason for the difference in expression between ERα and ERRα-mRNA is unclear, estrogen may regulates OPN expression through ERα and/or ERRα, either independently or in combination. Moreover, the decrease of OPN induced by removal of estrogen may increase urinary stones in postmenopausal women.
Asunto(s)
Receptor alfa de Estrógeno/genética , Hiperoxaluria/genética , Osteopontina/genética , Receptores de Estrógenos/genética , Animales , Células Cultivadas , Estradiol/análogos & derivados , Estradiol/farmacología , Femenino , Humanos , Hiperoxaluria/tratamiento farmacológico , Hiperoxaluria/metabolismo , Riñón/efectos de los fármacos , Riñón/metabolismo , Masculino , Ovariectomía , Progesterona/farmacología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas Wistar , Receptor Relacionado con Estrógeno ERRalfaRESUMEN
Epidermal cysts of the upper urinary tract are extremely rare. Only three cases have been reported in the published work written in English, Italian or German. We encountered a case of an epidermoid cyst in the ureter of a 72-year-old male. Findings on urine analysis and radiological examination were useful for establishing a correct diagnosis of epidermoid cyst of the urinary tract.
Asunto(s)
Quiste Epidérmico/diagnóstico , Enfermedades Ureterales/diagnóstico , Adulto , Anciano , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Alterations in intracellular Ca2+ ([Ca2+]i) are generally associated with cellular distress. Oxalate-induced cell injury of the renal epithelium plays an important role in promoting CaOx nephrolithiasis. However, the degree of change in intracellular free calcium ions in renal epithelial cells during oxalate exposure remains unclear. The aim of this study is to determine whether acute short-term exposure to oxalate produces morphological changes in the cells, induces a change in cytosolic Ca2+ levels in renal tubular epithelial cells and whether the application of extracellular glycosaminoglycans (GAGs) prevents these changes. Cultured Mardin-Darby canine kidney cells were exposed to oxalate, and changes in cytosolic Ca2+ were determined under various conditions. The effect of heparin and heparan sulfate (HS) during oxalate exposure was examined. The change in the GAG contents of the culture medium was also determined. Transmission electron microscopy (TEM) was performed for morphological analysis. The degree of change in cytosolic Ca2+ strongly correlated with oxalate concentration. Cytosolic Ca2+ levels decreased in parallel with an increase in the concentration of oxalate. However, this decrease was strongly inhibited by pretreatment with heparin or HS. TEM revealed cytoplasmic vacuolization, the appearance of flocculent material and mitochondrial damage after oxalate exposure. On the other hand, pretreatment with heparin or HS completely blocked these morphological changes. The present data suggest that acute exposure to a high concentration of oxalate challenges the renal cells, diminishes their viability and induces changes in cytosolic Ca2+ levels. Heparin and HS, which are known as potent inhibitors of CaOx crystallization, may also prevent oxalate-induced cell changes by stabilizing the cytosolic Ca2+ level.
Asunto(s)
Oxalato de Calcio/farmacología , Calcio/metabolismo , Citoprotección , Heparina/farmacología , Heparitina Sulfato/farmacología , Membranas Intracelulares/metabolismo , Riñón/metabolismo , Riñón/patología , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Perros , Líquido Extracelular/metabolismo , Riñón/efectos de los fármacos , Riñón/fisiopatología , Cálculos Renales/prevención & control , Microscopía ElectrónicaRESUMEN
A 36-year-old man was admitted to hospital due to right flank pain as a result of ureteral stones. He had been followed up for type 1 glycogen storage disease since the age of 11 years. He had four episodes of spontaneous stone birth during the previous 2 years, and each stone was composed mainly of calcium oxalate. Intravenous pyelography showed right hydronephrosis due to ureteral stones and bilateral multiple renal stones. We carried out transurethral ureterolithotripsy (TUL) on the right ureteral stones. The composition was a mixture of calcium oxalate and calcium phosphate. Laboratory evaluation demonstrated the association of distal renal tubular acidosis (RTA). These observations suggest that hypocitraturia and distal RTA are strongly correlated to recurrence of calcium nephrolithiasis. The patient's serum uric acid and urinary citrate excretion levels normalized after allopurinol and potassium citrate administration.
Asunto(s)
Acidosis Tubular Renal/complicaciones , Enfermedad del Almacenamiento de Glucógeno Tipo I/complicaciones , Cálculos Renales/etiología , Adulto , Calcio/análisis , Humanos , Cálculos Renales/química , MasculinoRESUMEN
OBJECTIVES: To examine the possibility of antegrade incisions at varying stricture lengths. We have developed a new method of using a ureteroscope and holmium:yttrium-aluminum-garnet (YAG) laser to make an antegrade incision without using a guidewire. Endoscopic internal urethrotomy involves the use of a guidewire or ureteral catheter that is passed through the stricture as an indicator for retrograde incision. METHODS: An antegrade incision was performed in 31 procedures for 28 patients with urethral strictures. We used a semirigid ureteroscope with an outer diameter of 6F at the tip. The ureteroscope was inserted into the urethra and passed through the stricture into the bladder under direct vision. The ureteroscope was pulled distally while an incision was made using the holmium:YAG laser at the 10-o'clock and 2-o'clock positions to a diameter of 17F. The endoscope was then replaced by a 17F panendoscope and an antegrade incision was similarly made up to 21F to 22F. RESULTS: An antegrade incision without the use of a guidewire was possible in all cases. Of the 31 incisions, restenosis appeared in 11 (35%). Of the 11 cases, re-incision was performed in 4 cases, and urethral sounding was conducted in the other 7 cases. Of the 4 re-incision cases, restenosis recurred in only 1 case. Of the 31 incisions, 23 (74%) were eventually successful. CONCLUSIONS: Antegrade incision using the narrow-diameter ureteroscope and holmium:YAG laser is a safe and easy method. This method is especially effective in cases of long strictures.
Asunto(s)
Terapia por Láser/métodos , Ureteroscopios , Estrechez Uretral/cirugía , Adulto , Anciano , Anciano de 80 o más Años , Humanos , Terapia por Láser/instrumentación , Persona de Mediana Edad , Recurrencia , ReoperaciónRESUMEN
We reported that expression of both HSPG and of bikunin are increased in calcium oxalate (CaOx) nephrolithic rat kidneys (lida et al., J. Am. Soc. Nephrol. 1999, Urol. Res. 1997). However, these findings were obtained from separate experiments. The present study evaluates whether levels of HSPG and bikunin expression differ in the rat kidney during calcium oxalate nephrolithiasis. Twenty-four male Wistar rats weighing 200-250 g were assigned to one of four groups (n = 6 each group) and administered with 0.5% ethylene glycol daily and 0.5 microgram of 1 alpha-OH-D3 every other day to induce CaOx nephrolithiasis. Animals were sacrificed 1 or 2 weeks later and both kidneys were excised. The cortex was separated from the medulla and papillary tips in the right kidney, then stored in liquid nitrogen for quantitative competitive-reverse transcription-polymerase chain reaction (QC-RT-PCR). The left kidney was fixed in 10% buffered formalin for histochemical studies. We assessed the variable gene expression of both HSPG and bikunin by QC-RT-PCR. Immunohistochemical analyses of left kidney tissue samples determined the localization of HSPG and bikunin. Normal rats serving as controls (n = 6 each) were also sacrificed and processed in the same manner as the experimental groups. QC-RT-PCR confirmed that HSPG and bikunin mRNA expression is significantly increased in nephrolithic kidneys (p < 0.05; Mann-Whitney test), and that medulla and papillary tips tended to express more mRNA of both. Immunohistochemical studies revealed that the production of HS and bikunin was increased in both the distal and proximal tubules of nephrolithic kidneys. These findings suggest that the increased expression of both HSPG and bikunin play an important role during calcium oxalate stone formation. In addition, this phenomenon might be associated with the progression of urothelial damage.
Asunto(s)
Oxalato de Calcio/metabolismo , Proteoglicanos de Heparán Sulfato/biosíntesis , Heparitina Sulfato/biosíntesis , Cálculos Renales/metabolismo , Riñón/metabolismo , Glicoproteínas de Membrana/biosíntesis , Inhibidor de la Tripsina de Soja de Kunitz , Animales , Proteoglicanos de Heparán Sulfato/genética , Heparitina Sulfato/análisis , Heparitina Sulfato/genética , Inmunohistoquímica , Masculino , Glicoproteínas de Membrana/genética , ARN Mensajero/análisis , Ratas , Ratas Wistar , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Regulación hacia ArribaRESUMEN
To develop a gene therapy for osteopathies, this study was conducted to establish a method of transferring the BMP gene, a bone formation factor, to cells and administering the cells with BMP expression to patients with osteopathies. Although virus vectors are frequently used for gene transfer, there has been reported a death case of gene therapy using the adenovirus vector. Therefore, various efforts have been made to prevent such complications. In the present study, we used electroporation by which gene transfer can be efficiently performed without inducing severe complications after electric perforation of the cell membrane. Human bone tissues were initially collected intraoperatively, and BMP-2 and Smad4 genes were cloned and integrated into GFP and DsRed plasmid vectors. Using in vitro electroporation, these plasmid vectors were transferred to the cultured chondrocytes (KTN-1) derived from human herniated intervertebral disk. Confocal laser microscopy revealed that the BMP gene was successfully transferred to the nucleus of chondrocytes in the presence of Smad. Since electroporation facilitated human gene transfer to the target cells, gene therapy using electroporation may facilitate individualized treatment for patients.
Asunto(s)
Condrocitos/ultraestructura , Electroporación/métodos , Técnicas de Transferencia de Gen , Microscopía Confocal/métodos , Factor de Crecimiento Transformador beta , Secuencia de Bases , Enfermedades Óseas/terapia , Proteína Morfogenética Ósea 2 , Proteínas Morfogenéticas Óseas/genética , Cartilla de ADN , Proteínas de Unión al ADN/genética , Terapia Genética , Humanos , Técnicas In Vitro , Rayos Láser , Proteína Smad4 , Transactivadores/genéticaRESUMEN
We have reported that heparan sulfate (HS)/heparan sulfate proteoglycan (HSPG, syndecan-1) expression significantly increased in the rat kidney during calcium oxalate (CaOx) nephrolithiasis. Although the exact role of syndecan still remains unclear, HS/syndecan-1 is thought to have some important role in the CaOx crystal formation. Mardin-Darby canine kidney (MDCK) cells are most commonly used in kidney stone research. It was reported that MDCK cells do not express syndecan-1. In the present study, we established a novel MDCK cell line (KIC-synd-1) that expressed the human syndecan-1 gene. In this cell line, we confirmed stable expression of both sndecan-1 gene and core protein. Immunohistochemical study revealed positive staining of syndecan-1 monoclonal antibody in the basolateral and cytosolic area of the KIC-synd-1 cells. We also investigated the composition of glycosaminoglycans (GAGs) side chains in MDCK cells and KIC-synd-1 cells by using high-performance liquid chromatography (HPLC). Four types of HS chains were identified in both cells as follows; delta diHS-NS, delta diHS-6S, delta diHS-diS1, delta diHS-diS2. Increased production of delta diHS-NS and delta diHS-diS2 were shown in KIC-synd-1 cells compared with production in MDCK cells (p < 0.05). In contrast, only a small amount of delta diHS-6S and delta diHS-diS1 was contained in both cell lines. Total amount of HS was significantly increased in the KIC-synd-1 cells compare with that in the wild type MDCK cells (p < 0.05). Scanning electron microscopy revealed no significant difference between cell surface of wild type MDCK cells and that of KIC-synd-1 cells in normal conditions. However, calcium oxalate crystal attachment was apparently decreased in the KIC-synd-1 cells. These results suggested that cell surface HS/syndecan-1 has preventive role for calcium oxalate nephrolithiasis via creation of a charge barrier against COM crystal attachment.