Eicosanoids in the pathogenesis of the functional and structural alterations of the kidney in diabetes.

Diabetes mellitus alters the cellular production of eicosanoids in a number of tissues, including the kidney, and these agents have in turn been implicated in the pathogenesis of diabetic nephropathy. As delineated in the streptozotocin diabetic rat (SDR) model, a preferential enhancement of glomerular synthesis of the vasodilatory prostaglandins (PGs) PGE2 and PGI2 with concurrent smaller increases in thromboxane (TX)A2 occurs within 1 week after induction of diabetes. This early alteration in glomerular synthesis of eicosanoids in the SDR has been linked to glucose-induced activation of the glomerular protein kinase C signalling system that enhances phospholipase A2 activity and, therefore, release of membrane-bound arachidonic acid for oxygenation. The preferential increase in glomerular production of vasodilatory PGs may contribute to the glomerular hyperfiltration that is characteristic of early diabetes. After more prolonged (months) diabetes in the SDR, glomerular generation and urinary excretion of thromboxane (TX) are preferentially enhanced. Studies with selective inhibitors of TX synthesis in the SDR have implicated this eicosanoid in the pathogenesis of both albuminuria and glomerular structural changes (basement membrane thickening and mesangial matrix expansion). Direct stimulation of matrix protein production has been demonstrated in cultured mesangial cells in response to both TX and high ambient concentrations of glucose. The actions of TX and glucose on mesangial cell matrix production appear to be interactive, with each signalled through distinct pathways of protein kinase C activation.

Effects of aspirin on 1,2-dimethylhydrazine-induced colonic carcinogenesis.

The influence of aspirin (ASA) on 1,2-dimethylhydrazine (1,2-DMH)-induced colonic carcinogenesis was examined in weanling Sprague-Dawley rats. The incidence of adenocarcinomas in response to a single dose of 1,2-DMH was reduced 60% in rats receiving ASA for 1 week before and after the carcinogen. However, ASA had no effect on tumor incidence when initiated 4 weeks after a single dose of 1,2-DMH and continuing until the animals were killed at 36 weeks. The doses of ASA employed suppressed by 95% or more ex vivo colonic prostaglandin E2 (PGE2) production and reduced colonic mucosal cAMP levels in both rats exposed to 1,2-DMH and in age-matched controls. Proliferative activity of colonic mucosa as assessed from tritiated thymidine ([3H]dThd) incorporation into mucosal DNA was increased at 1 week but suppressed by 36 weeks after 1,2-DMH exposure. ASA significantly increased colonic mucosal DNA synthesis, suppressed colonic PGE2 production and reduced mucosal cAMP levels at both 1 and 36 weeks in rats given the 1,2-DMH vehicle. However, ASA failed to alter the enhanced mucosal DNA synthesis observed at 1 week or the suppressed DNA synthesis observed at 36 weeks after a single dose of 1,2-DMH, despite significant inhibition of colonic PGE2 production and reduction in mucosal cAMP levels by ASA. Treatment of rats for 1 week with ASA significantly inhibited basal and arachidonate stimulated decomposition of the 1,2-DMH intermediary metabolite methylazoxy-methanol, assessed ex vivo in colonic mucosal homogenates. Thus, while other mechanisms are not excluded, suppression of 1,2-DMH induced colonic carcinoma by concurrent administration of ASA may be linked in part to altered metabolic activation of this carcinogen via cyclooxygenase-dependent co-oxidation. By contrast, the previously reported suppression of the promotional phase of colonic carcinogenesis in rats by the delayed introduction of cyclooxygenase inhibitors may not be linked to inhibition of local colonic prostanoid production, since (i) inhibition of colonic prostanoid synthesis by ASA did not mimic this antipromotional effect, and (ii) the doses of non-steroidal anti-inflammatory drugs employed in some earlier studies may not significantly inhibit colonic prostanoid synthesis.

Contribution of platelet thromboxane production to enhanced urinary excretion and glomerular production of thromboxane and to the pathogenesis of albuminuria in the streptozotocin-diabetic rat.

Previous studies have demonstrated that urinary thromboxane B2 (TXB2) excretion (UTXB2) and glomerular production of TXB2 are enhanced in experimental diabetes and that selective inhibitors of TX synthesis prevent or delay the development of albuminuria. The present study was conducted to examine the contribution of platelet TXB2 production to the enhancement of UTXB2 and glomerular TXB2 production and to the pathogenesis of albuminuria in the partially insulin-treated moderately hyperglycemic (blood glucose, 200 to 400 mg/dL) streptozotocin-diabetic rat (SDR). Treatment of control rats or of SDR with diabetes of 5 months' duration with antiplatelet serum for 4 consecutive days reduced circulating platelet counts and serum TXB2 generation, an index of platelet cyclooxygenase activity, by 80% or greater, but reduced UTXB2 excretion by only 30%. UTXB2 and glomerular production of TXB2 of thrombocytopenic SDR remained markedly elevated compared with corresponding values from age-matched thrombocytopenic or platelet-replete, nondiabetic controls. Similarly, treatment of rats for 180 days with a dose of aspirin (ASA), which selectively inhibited platelet versus renal cyclooxygenase activity, reduced UTXB2 of both SDR and controls by 25% to 35%. The absolute reductions in UTXB2 induced by either ASA or thrombocytopenia in SDR were significantly greater than the absolute decrements in corresponding controls, suggesting that increased platelet TXB2 production in SDR may contribute to the enhanced UTXB2. However, as in the thrombocytopenic SDR, UTXB2 and glomerular production of TXB2 of SDR treated with ASA remained clearly above corresponding control values. Moreover, chronic ASA treatment failed to prevent the development of albuminuria in SDR.(ABSTRACT TRUNCATED AT 250 WORDS)

Calcium and prostaglandin E2 in renomedullary interstitial cells.

Renomedullary interstitial cells cultured from the Dahl salt-resistant rat have higher levels of basal cytosolic calcium and prostaglandin E2 and are more responsive to vasopressin than interstitial cells from the Dahl salt-sensitive rat. We examined the potential role of inositol phospholipid hydrolysis in mediating these differences. Vasopressin-induced increases in labeled inositol phosphates were enhanced in renomedullary interstitial cells from Dahl salt-resistant compared with those from salt-sensitive rats. Addition of neomycin reduced basal production of labeled inositol phosphates and abolished the increase in inositol phosphates induced by vasopressin. Neomycin also prevented the peak decline pattern in cytosolic Ca2+ seen with vasopressin but did not influence basal cytosolic Ca2+. In the presence of neomycin, vasopressin induced a modest but prolonged increase in cytosolic calcium. In contrast to its marked effects on inositol phosphate production, neomycin was without effect on basal or vasopressin-responsive prostaglandin E2 production. Moreover, basal and vasopressin-induced increases in cytosolic Ca2+ remained higher in renomedullary interstitial cells from Dahl salt-resistant versus those from salt-sensitive rats exposed to neomycin. The results do not support a requirement for phospholipase C-induced inositol phospholipid hydrolysis in the mediation of vasopressin actions on prostaglandin E2 production by renomedullary interstitial cells and imply that the differences in cytosolic Ca2+ and prostaglandin E2 seen in these two cell lines are not related to differences in inositol phospholipid metabolism.

Suppression of urinary albumin excretion in diabetic rats by 4'(imidazol-1-yl) acetophenone, a selective inhibitor of thromboxane synthesis.

Thromboxane contributes to the regulation of glomerular hemodynamics in experimental models of diabetes and has been implicated as mediator in some models of glomerular injury. In the present study we examined urinary albumin, protein, and thromboxane B2 (TXB2) excretion during the 170 days after induction of diabetes by injection of streptozotocin in insulin-treated moderately hyperglycemic (200 to 400 mg/dl glucose) rats (SDRs). The effects of a thromboxane synthesis inhibitor, 4'-(imidazol-1-yl)acetophenone (TXI) (100 mg/kg/day) on these parameters were also assessed. Urinary TXB2 and albumin excretion in SDRs was not different from that in normal rats between 7 and 90 days but were three times higher than normal in SDRs at 125 and 170 days after induction of diabetes. In SDRs, urinary protein excretion was higher than in controls at 170 days but not at earlier time points. Inulin clearance (CIn) of SDRs was significantly higher than control values at 7 and 90 days and was not influenced by TXI during this period. At 170 days CIn was not significantly different in SDRs and normal rats. By contrast, albumin clearance (CAIb) and fractional CAIb were elevated in SDRs when compared with those values in normal rats. Treatment of SDRs with TXI for 170 days completely prevented the rise in urinary TXB2, albumin, and protein excretion, as well as the rise in fractional CAIb, but did not alter prostaglandin E2 (PGE2) excretion. TXI also increased CIn in SDRs to levels that were significantly higher than normal at 170 days. TXI had no significant effect on urinary PGE2, TXB2, albumin, or protein excretion or on CIn in normal rats and did not influence blood pressure or blood glucose in normal rats or SDRs. The results suggest a role for thromboxane in the mediation of albuminuria in the SDR.