To evaluate the ultrastructural changes and the mechanism causing liver injury by lead, light and electron microscopic(LM and EM) examination using Timm sulphide silver method(TSM) was done. Sprague-Dawley rats were divided into a control and 3 experimental groups. The experimental groups were orally administered 0.5% lead acetate(LA). Group 1 received a one time dose of 10 ml of LA by gastric intubation. Groups 2 and 3 continuously received LA instead of drinking water. The control group was composed of 3 rats in each group which did not receive any treatment. Rats of group 1, 2 and 3 and control were sacrificed at 1/2, 1, 1 1/2 hours, 2 days, and at 1, 2, 4, 6 and 8 weeks later, except group 3. Before sacrifice, they were perfused with 0.1% sodium sulphide and 2.5% glutaraldehyde through the abdominal aorta for TSM. The liver was taken for LM and EM examinations. Blood lead concentration began to increase from the 2nd day up to 3.29 microgram/ml at 2nd week, and the urinary delta-ALA level showed a steady increase from the 2nd day. LM and EM examination of liver revealed that absorbed lead granules in group 1 were transported into sinusoidal spaces, Kupffer cells, and the hepatocytes within 1 hour and then disappeared 1/2 hour thereafter. In group 2 deposited lead was found in the hepatocytic cytosol bound to mitochondria. That in turn inhibited mitochondrial respiration with resultant mitochondrial swelling at the 1st week and thereafter at 6th week myelin figure formation and condensation of mitochondria, and peroxisomes were increased at 8th week. Based on these results it can be concluded that a transient intake of subletal dose of LA is biotransformed completely by periportal hepatocytes within 1 1/2 hours, but excessively accumulated lead can induce liver cell injury due to lipid peroxidation of membrane by direct toxic effect of lead and by products of lipid peroxidation. We postulate that lead acetate triggers presumably primarily mitochondrial membrane injury and then other organellar changes may play a role in disturbance of a network of interacting of key events capable of causing cell death.
Rats ; Animals

The purpose of this study was to investigate the morphologic changes of the bile canaliculi and its associated structures of the liver induced by common bile duct ligation(CBDL) in the rat. The canalicular surface and lateral surface of the dry-fractured hepatocytes was studied with scanning electron microscopy at 1~6 weeks post ligation. The first week after CBDL, the bile canaliculi were dilated. The microvilli were increased in number and the lumens contained granular materials After 2 weeks or more, the bile canaliculi were dilated to a variable degree, and with irregularity, measuring from 1.5 to 5 micrometer in diameter, and in the advanced stage, the canaliculi showed blunting and the disappearance of microvilli. Some canaliculi had sprouting side branches. At 4~6 weeks post-ligation, the lateral surface of the hepatocytes also showed some irregularity and a tortuous appearance, and numerous small sized microvillous projections were formed. The tubular structures of the proliferated SER distributed adjacent to the lateral surface of the hepatocytes, and the direct connection of a tubular structure and the cytoplasmic membrane was observed. These results suggest that the deformity and loss of microvilli of bile canaliculi reflect the disturbance of bile secretion from the hepatocytes. And prolonged obstruction of bile flow may result in bile excretion via the lateral surface of hepatocytes.
Rats ; Animals

No abstract available.
Animals ; Rats*

No abstract available.
Animals ; Rats*

The cholestasis are defined as blockade or secretory distrubance of bile and appearance of bile in hepatocytes, Kupffer cells and biliary passages, usually associated with dilated bile canaliculi. Intra-and extraheptic cholestasis were induced by 17-ethinyl estradiol, or chlorpromazine hydrochloride and by ligation of bile duct to investigate the mechanism of the hepatic injury, ultrastructural changes of liver and alterations of liver function. The results obtained were as follows. 1) Functional and histological changes of intra-and extrahepatic cholestasis differed in various experimental groups. The liver weight is increased in 17-ethinyl estradiol treated group and ligation of bile duct group (5.6+/-0.15, P<0.001, 5.3+/-0.19 gm/100 gm body weight, P<0.001). The common features of intra-and extrahepatic cholestasis were double membrane bounded amorphous vesicular material infiltrations in the cytoplasm of hepatocyte, partial loss of microvilli of bile canaliculi, anf focal thickening of pericanalicular ectoplasm on electron microscopy. 2) Intrahepatic cholestasis induced by 17-ethinyl estradiol show significantly increased serum level of alkaline phosphatase and total bile aicd (134.0+/-16.82 IU/L, 29.5+/-4.68 umol/l). Kupffer cell proliferation and focal cytoplasmic degradation with myelin figures are characteristic features on electron microscopy. Chlorpromazine hydrochloride induced intrahepatic cholestasis show increased serum level of AST, ALT, Cholesterol and bilirubin (156.9+/-11.32, 49.0+/-2.83 IU/L, 59.3+/-6.73 mg/dl, 1.8+/-.043 mg/dl). Inflammatory cell infiltration, chiefly lymphocytes and esoinophils are seen in periportal area. Prominent vesiculation and vacuolations of smooth endoplasmic reticulum are characteristic feature on electron microscopy. 3) Extrahepatic cholestasis induced by ligation of bile duct show increase serum level of AST, ALT, GGT, cholesterol, total bile acid, and bilirubin (290.2+/-50.24, 171.5+/-47.17, 159.3+/-24.54, 33.7+/-1.47 IU/L, 86.6+/-9.18 mg/dl, 246.6+/-27.34 umol/l, 13.9+/-0.83 mg/dl). Light microscopically, morphologic alterations are feathery degeneration of hepatocytes, proliferation of bile ducts, bile infarct and prominent intracytoplasmic lipid droplets. Electron microscopically, electron dense acidophilic body, bile casts and complete loss of microvilli are seen in dilated bile canaliculi. Also noted are hypertrophy of cannalicular ectoplasm. Finely granular materials are infiltrated in degenerative cytoplasm.
Rats ; Animals

No abstract available.
Animals ; Rats*

Chronic deposition of uric acid in the kidney can lead to progressive tubulointerstitial injury with granuloma formation. We hypothesized that uric acid crystal deposition may induce granuloma formation by stimulating local expression of macrophage migration inhibitory factor(MIF), which is a known mediator of delayed type hypersensitivity(DTH). A model of acute uric acid nephropathy was induced in rats by the administration of oxonic acid (an inhibitor of uricase) together with uric acid supplements. Kidney tissue examined at 35 days showed widespread tubulointerstitial damage with intratubular uric acid crystals deposition and granuloma formation. Tubules within the areas of granuloma showed a six-fold increase in MIF mRNA compared to uninvolved areas by in situ hybridization. Moreover, the areas of increased MIF mRNA expression correlated with sites of dense accumulation of macrophages and T cells. Control rats fed a normal diet had no discernible evidence of renal disease by routine light microscopy and minimal tubular expression of MIF mRNA and protein. These data suggest that intrarenal granulomas in urate nephropathy may be the consequence of a crystal induced DTH-like reaction mediated by MIF.
Rats ; Animals

Potassium(K+) balance is achieved by the control of urinary K+ excretion and by the control of K+ absorption from the digestive tract. It has been established that chronic potassium depletion is associated with a remarkable hypertrophy of the outer medullary collecting duct of the kidney. But, there are no morphological studies regarding the intercalated cells during the chronic changes of potassium diet. Electron microscopy was performed to observe the morphological alterations of the intercalated cell of the entire collecting duct in response to chronic changes of potassium diet in rat kidney. By electron microscopy, the characteristic features of normal type A intercalated cell of the cortical collecting duct included numerous micro-projections of the apical plasma membrane, complicated basal infolding, apical cytoplasmic tubulovesicles, evenly distributed mitochondia, and centrally located nucleus. In potasium-depleted type A intercalated cell, microprojections of the apical plasma membrane were increased in length and number, basal infoldings were uncomplicated, tubulovesicles were almostly disappeared, and mitochondria were increased in number. Type A intercalated cell of potassium-loading after restriction was found to be almost normal except longer microprojections and increased mitochondria. The characteristic features of normal intercalated cell of the outer medullarycollecting duct(OMCD) included relatively short micro-projections of the apical plasma membrane, uncomplicated basal infoldings, apical cytoplasmic tubulovesicles, and apically distributed mitochondia. In comparison with normal, potassium-depleted intercalated cell of OMCD was hypertrophy, microprojections of the apical plasma membrane were increased in length and number, basal infoldings were complicated, tubulovesicles were almost disappeared, mitochondria were increased in number, and several lysosomes were appeared. Intercalated cell of OMCD of potassium-loading after restriction was found to be almost normal except increased cell size, longer microprojections, and increased mitochondria and lysosomes compared to control. The characteristic features of normal intercalated cell of the inner medullary collecting duct (IMCD) included very short and scant microprojections of the apical plasma membrane, uncomplicated basal infoldings,apica cytoplasmic tubulovesicles, evenly distributed mitochondia, and some lysosomes. In potasium-depleted intercalated cell of IMCD, cell size was prominently increased, microprojections of the apical plasma membrane were increased in length and number, basal infoldings were complicated, tubulovesicles were almostly disappeared, and mitochondria were increased in number. Intercalated cell of IMCD of potassium-loading after restriction was found to be almost normal except increased cell size and increased microprojections in number and length compared to control. These results suggest that intercalated cells adapt through morphological changes to preserve potassium balance during chronic changes of potassium diet.
Rats ; Animals

Diuretics are natriuretic agents which inhibit sodium reabsorption at their major site of action on the renal tubules and increase the excretion of sodium and combined anions. Increment of urine volume is the secondary to the natriuretic effects. Diuretic resistance occurs that threshold dose of diuretics is higher than that of other patients. It is frequently manifested among the edematous patients such as those with the nephrotic syndrome. Prolonged use of diuretics decreases the natriuretic effect of diuretics, which is called diuretic tolerance. This is important adaptations of distal nephron segment. To elucidate the mechanism of diuretic resistance, 1 mg of bumetanide was given to the nephrotic syndrome(NS) patients group and control group, respectively. The peak plasma concentration was delayed in NS patients. The proportion of urine free bumetanide for 24 hours was 73% in NS patients but 100% in control group. The ratio of urine volume and amount of Na+ and Cl - for 24 hours to the total and free urine bumetanide was decreased in NS patients. The study suggests that pharmacokinetic and pharmacodynamic changes of the diuretics induce the diuretic resistance. To determine the additive diuretic effect of albumin to the action of the furosemide, 160 mg of furosemide was administered intravenously with albumin in NS patients. Simultaneous infusion of albumin and furosemide did not enhance the diuretic effect of furosemide pharmacodynamically and pharmacokinetically. Albumin preinfusion 30 minutes before furosemide administration potentiates the diuresis, but natiuresis and pharmacokinetics were not changed. Semiquantitative immunoblotting of rat kidneys was carried out to investigate whether chronic diuretics adminstration induces changes in the abundance of Na+ transporters in distal nephron. Furosemide infusion increased cortical and outer medullary abundances of Na+-Cl- cotransporter(TSC) and all 3 subunits of the epithelial sodium channel(ENaC). Hydrochlorothiazide infusion increased abundances of some kinds of subunits of ENaC. These increases in the abundances of Na+ transporters may account for the generation of diuretic tolerance.These data suggest that to overcome the diuretic resistance or tolerance, diuretic dose increment over the threshold level and more frequent administration of the diuretics are recommended. Diuretic combinations are also proposed. Addition of albumin to augment the diuretics effect ought to be considered cautiously.
Rats ; Animals

No abstract available.
Animals ; Rats*