摘要:

In this review the characteristics of established renal and intestinal epithelial cell lines are described by summarizing the accumulated literature about specific properties retained by the cells in tissue culture. Furthermore, brief examples are given for the use of cultured epithelia as model systems to study epithelial transport and metabolic functions, epithelial cell polarity, and aspects of the differentiation and maturation of epithelia by physiological, biochemical and genetic, or cell and molecular biological approaches.

ISSN:1420-4096    

DOI:10.1159/000173147

出版商:S. Karger AG    

年代:1988

数据来源: Karger

摘要:

The rat kidney was separated into the outer cortex, inner cortex, outer medulla and inner medulla or papilla, and the distribution of angiotensin-converting enzyme (ACE) in response to consecutive administrations of captopril was studied. In normal animals, the ACE activity in the inner cortex and outer medulla was about 10 and 3 times higher than in the outer cortex, respectively, and the activity in the inner medulla was much the same as that in the outer cortex. Captopril was given in doses of 30 and 100 mg/kg/day for 7 days, and the renal ACE activity on the 8th day was examined. Since it was assumed that the ACE activity might be lower than the full activity when the tissue contained captopril, diamide, a sulfhydryl oxidant, was used to eliminate the effect of any captopril remaining in the tissue extracts. However, this compound at concentrations over 1 mM enhanced the activity of ACE itself to about twice the value when assayed without diamide. Thus, oxidation with sufficient concentrations of diamide resulted in an enhancement of a maximal activity of ACE. When all samples were pretreated with 10 mM diamide and the ACE activity determined, captopril caused a dose-related increase in the ACE activity, but only in the inner cortex. We conclude that ACE locates predominantly in the inner cortex of the rat kidney and it is this area which has an altered ACE activity in response to captopril.

ISSN:1420-4096    

DOI:10.1159/000173148

出版商:S. Karger AG    

年代:1988

数据来源: Karger

摘要:

We have found that arginine vasopressin (AVP) (10 pg/ml) stimulates urinary kallikrein in the isolated erythrocyte perfused rat kidney. (In this model, perfusate flow rate approximates blood flow rates in vivo and morphology is normal.) Urinary kallikrein excretion rose from 6.9 ± 0.8 to 14.9 ± 2.4 ng/min 20 min after the addition of AVP to the perfusate, and then fell towards baseline levels over the next 30 min. 1-Desamino-8-D-AVP (8 pg/ml) caused a comparable increase in kallikrein excretion. Prostaglandin synthesis inhibition with indomethacin did not alter the stimulatory effect of AVP on kallikrein excretion. Parathyroid hormone 1–34 (144 ng/ml) and calcitonin (102 ng/ml) also increased urinary kallikrein. Kallikrein excretion rose from 9.1 ± 2.0 to 24 ± 4.5 ng/min in response to calcitonin and from 8.3 ± 1.6 to 43.7 ± 3.4 ng/min following the addition of parathyroid hormone to the perfusate. Kallikrein was found to accumulate in the perfusate in a linear fashion. Based on the slope of the relationship between perfusate kallikrein and time, the rate of release of kallikrein into the perfusate was estimated to be 0.79 ng/min in control kidneys. The rate of release of kallikrein into the perfusate in kidneys treated with AVP was the same (0.74 ng/min). Thus while kallikrein is released into the perfusate, this process is not influenced by AVP. In conclusion, AVP stimulates release of kallikrein into the urine (but not the perfusate) independently of systemic events. The effect of AVP is not mediated by prostaglandins. This effect of AVP is mediated via stimulation of the V2 receptor and also occurs in response to two other hormones (calcitonin and parathyroid hormone) that are known to stimulate adenyl cyclase in the rat distal

ISSN:1420-4096    

DOI:10.1159/000173149

出版商:S. Karger AG    

年代:1988

数据来源: Karger

摘要:

We have examined the effects of 25 min of ischemia in the isolated erythrocyte-perfused rat kidney (IEPK). We have previously shown that, in this model, perfusate flow rate is close to blood flow rates in vivo and morphology is normal. The functional and morphological consequences of both warm ischemia (at 37°C) and ischemia induced during mild hypothermia (27°C) were compared. (1) Warm ischemia resulted in a 51% increase in renal vascular resistance (RVR) during the reflow period, while glomerular filtration rate (GFR) was reduced to 24% of control levels. (2) Kidneys subjected to warm ischemia showed marked morphological damage localized to the proximal tubule. There was dilatation of the proximal segments and widespread loss of the proximal brush border due both to shedding into the lumen and interiorization into the cell. In contrast to the proximal tubular damage, the cells of the medullary thick ascending limb segments were intact. However, the lumena of many of these segements were filled with cytoplasmic blebs and necrotic cell debris. There was also pronounced vascular congestion of the capillary plexus in the inner stripe of the outer medulla. (3) Hypothermia to 27°C resulted in almost complete protection against ischemic injury: RVR and GFR were not different from control values. Also, kidneys subjected to cold ischemia showed only isolated areas of mild brush border damage; no evidence of tubular obstruction or vascular congestion was present. (4) Thus, warm ischemia in the IEPK results in functional and morphological effects comparable to those found in vivo. Post-ischemic vasoconstriction as well as medullary congestion occur in the absence of systemic hormones and renal nerves. These consequences of ischemia are prevented by modest hypotherm

ISSN:1420-4096    

DOI:10.1159/000173150

出版商:S. Karger AG    

年代:1988

数据来源: Karger

摘要:

We have tested whether increased Ca++ and Mg++ concentrations have an effect on transepithelial voltage (PDte) and transepithelial resistance (Rte) in isolated perfused cortical thick ascending limbs (cTAL) of rabbit kidney. The divalent cations added at 2.5, 5.0 and 10.0 mmol · 1-1 to the lumen or peritubular bath perfusate led to a concentration-dependent increase in Rte. The maximal response in Rte was observed between 5 and 10 mmol · 1-1. No significant change in active transepithelial potential difference (PDte) was observed. The increase in Rte still occurred when the transcellular current was reduced by Ba++ (3 mmol · 1-1) added to the lumen perfusate. This suggests that the increase in Rte caused by Ca++ and Mg++ is due to a modification of the paracellular shunt pathway. In the absence of active transport, i.e. when furosemide (5 · 10-5 mol · 1-1) was added to the lumen perfusate, Ca++ and Mg++ reduced the transepithelial diffusion potential generated by a NaCl gradient established across the epithelium, and thus produced a reduction of the relative permeability for Na+ over Cl- (PNa+/Pcr) of the paracellular shunt pathway. This indicates that divalent cations increase Rte by reducing the sodium permeability of the tight junctions. The observed Ca++ and Mg++ induced reduction of the sodium permeability of the paracellular pathway corresponds to a decrease in net Na+ reabsorption by 5–10%. Since it has been demonstrated that peptide hormones such as parathyrin (PTH) modulate divalent cation and NaCl reabsorptions, in a second series of experiments we tested the effects of PTH (2–20 USP · 1-1) and dbcAMP (10-3 mol · 1-1) on PDte and Rte of isolated perfused cTAL segments of rabbit nephron. Neither Rte nor PDte were affected by PTH

ISSN:1420-4096    

DOI:10.1159/000173151

出版商:S. Karger AG    

年代:1988

数据来源: Karger

摘要:

Although marked alterations in temperature often accompany ischemic, acute renal failure (ARF), the effects of altered temperature on renal structure and function have received little attention. In the present investigation, isolated rat kidneys perfused at 41°C had extensive tubular damage and decreased function compared to kidneys perfused at 37°C. In contrast, kidneys perfused at 30°C had less tubular damage, and better function, than kidneys perfused at 37°C. Increased temperature caused a 50% reduction in renal ATP (0.46 ± 0.04 µM/100mg tissue protein, 37°C, vs. 0.26 ± 0.03 µM/100 mg tissue protein, 41°C; p < 0.05). The decreased ATP occurred despite reduced sodium reabsorption (129 ± 8 µM/min/g, 37°C, vs. 65 ± 12µMmin/g, 41°C, p < 0.05) and normal renal oxygen consumption (QO2). These results suggest that increased temperature may cause an uncoupling of QO2 and sodium chloride transport, and an increase in nontransport mediated, basal metabolic rate may result in depleted cellular ATP levels and renal tub

ISSN:1420-4096    

DOI:10.1159/000173152

出版商:S. Karger AG    

年代:1988

数据来源: Karger

摘要:

Because adenosine plays a role in the regulation of glomerular filtration rate and of the release of renin, we examined the possibility of a local source for this mediator. We found that rat cultured glomerular mesangial cells converted 5’-AMP into adenosine. The properties of the enzyme involved in the reaction were those of an ecto-5’nucleotidase: (1) the products of the reaction were generated in the extracellular fluid although no 5’-nucleotidase was released by the cells into the medium; (2) identical activities were found for cultured cells in situ and sonicated cells; (3) the diazonium salt of sulfanilic acid which is a nonpenetrating reagent inhibited up to 75% of the enzyme activity. Ecto-5’-nucleotidase activity of intact cells obeyed Michaelis-Menten kinetics. Apparent Km for 5’-AMP was 0.32 mM. 5’-UMP was a strictly competitive inhibitor. ADP exerted a very powerful inhibitory effect and behaved also as a competitive inhibitor. ATP was inhibitory both by increasing Km and by decreasing Vmax. Ecto-5’-nucleotidase was active in the absence of divalent cations. However, Mg2+, Ca2+, Co2+ and Mn2+ were stimulatory. Zn2+ and Cu2+ suppressed the activity. Concanavalin A, a plant lectin, was markedly inhibitory, suggesting that a glycoprotein moiety was necessary to express enzyme activity. Ecto-5’-nucleotidase activity was not modified during phagocytosis of serum-treated zymosan by mesangial cells. Rat cultured glomerular epithelial cells exhibited a 5’-nucleotidase activity which was 4 times lower than that of the mesangial cells in primary culture. Activity was also present in freshly isolated glomeruli and total cortex, but at lower concentrations than in mesangial cells. These findings suggest that ecto-5’-nucleotidase of mesangial cells may be the main source of adenosine within the glomeruli. Therefore, activity of this enzyme should be essential in the regulation of the glomerul

ISSN:1420-4096    

DOI:10.1159/000173153

出版商:S. Karger AG    

年代:1988

数据来源: Karger

摘要:

The glomerular filtration rate (GFR), effective renal plasma flow (ERPF), systolic blood pressure (SBP) and urinary protein excretion (UPV) were determined in 12-week-old male rats of the spontaneously hypertensive Fawn-Hooded (FH) strain. These data were compared with those of either age-matched or weight-matched male, normotensive Wistar Albino Glaxo (WAG) rats. The GFR was significantly higher in FH rats than in both WAG control groups. In contrast, the ERPF did not differ between the FH and WAG rats. Thus, a higher filtration fraction was present in the FH rats. As no differences were found in the total number of glomeruli per kidney comparing FH and WAG rats, the high GFR was not due to an increase in the number of glomeruli. The SBP and the UPV were significantly higher in FH rats than in WAG rats. To our opinion, the arterial hypertension associated with glomerular hyperfiltration proteinuria suggests the presence of glomerular hypertension in FH rats.

ISSN:1420-4096    

DOI:10.1159/000173154

出版商:S. Karger AG    

年代:1988

数据来源: Karger