Alterations in Nitric Oxide Activity and Sensitivity in Early Streptozotocin-Induced Diabetes Depend on Arteriolar Size

Changes in NO activity may play an important role in the early increase in microvascular flow that has been implicated in the pathogenesis of diabetic microangiopathy. We assessed, in the in situ spinotrapezius muscle preparation of 6 weeks' streptozotocin-diabetic rats (n = 6) and of agematched controls (n = 8), basal inside diameters of A2–A4 arterioles and the reactivity to topically applied acetylcholine and nitroprusside, before and after NG-nitro-L-arginine. In diabetic rats, cholinergic vasodilatation in A2–A4 arterioles was intact. Basal diameter in A3 and A4 arterioles was significantly higher in streptozotocin-diabetic rats. The increased basal diameter in A3 arterioles was partially due to an increased contribution of NO to basal diameter. The response to nitroprusside was impaired in streptozotocin-diabetic rats in A2, but not in A3 and A4 arterioles. Thus, this study shows that NO activity and sensitivity are altered after 6 weeks of streptozotocin-induced diabetes. These streptozotocin-induced changes are anatomically specific and, for arterioles, depend on their position within the vascular tree

Increased leukocyte rolling in newly formed mesenteric vessels in the rat during peritoneal dialysis

OBJECTIVE: Long-term peritoneal dialysis (PD) is associated with the development of functional and structural alterations of the peritoneal membrane. The present study reports the effects of chronic exposure to PD fluid on mesenteric leukocyte-endothelium interactions, using intravital video microscopy. METHODS: Rats (n = 7) received 10 mL lactate-buffered 3.86% glucose-containing PD fluid daily during a 5-week period via a subcutaneously implanted mini access port that was connected via a catheter to the peritoneal cavity. In a first control group (n = 8), catheters were implanted but no fluid was instilled; a second control group (n = 8) remained untreated. The number of rolling and adherent leukocytes as well as blood flow and other fluid dynamic variables were analyzed in mesenteric postcapillary (diameter 10-25 mu) and collecting (diameter 26-40 mu) venules. Neovascularization was semiquantitatively assessed after inspection of video images and by light and electron microscopy. Using FITC-labeled albumin, microvascular leakage was examined. RESULTS: Rats exposed to PD fluid showed a more than twofold increase in the number of rolling leukocytes (p < 0.01); the number of adherent leukocytes was not changed. Furthermore, exposure to PD fluid induced severe neovascularization in rat mesentery. No microvascular leakage was observed in the various groups. The observed differences could not be explained by differences in systemic or local hemodynamic parameters or peripheral leukocyte counts, but is most likely associated with new vessel formation. CONCLUSIONS: Exposure of rat peritoneal membrane to conventional PD fluid for 5 weeks affected local leukocyte-endothelium interactions. In addition, severe angiogenesis was induced, whereas microvascular permeability remained unaltered

Better preservation of the peritoneum in rats exposed to amino acid-based peritoneal dialysis fluid

BACKGROUND: Glucose-containing peritoneal dialysis fluids (PDF) show impaired biocompatibility, which is related partly to their high glucose content, presence of glucose degradation products, low pH, and lactate buffer, or a combination of these factors. In a rat chronic peritoneal exposure model, we compared effects of an amino acid-based PDF (AA-PDF) with a glucose-containing PDF on the peritoneal microcirculation and morphology. METHOD: Two groups of rats received 10 mL of either fluid daily for 5 weeks via peritoneal catheters connected to implanted subcutaneous mini vascular access ports. Leukocyte-endothelium interactions in the mesenteric venules were investigated by intravital microscopy. Quantification of angiogenesis and fibrosis and inspection of the mesothelial cell layer were performed by light and electron microscopy. RESULTS: Daily exposure to glucose-containing PDF resulted in a significant increase in the number of rolling leukocytes in mesenteric venules, whereas instillation of AA-PDF did not change the level of leukocyte rolling. Glucose-containing PDF evoked a significantly higher number of milky spots in the omentum, whereas this response was significantly reduced in animals exposed to the AA-PDF (p < 0.02). Chronic instillation of glucose-containing PDF induced angiogenesis in various peritoneal tissues, accompanied by fibrosis in the mesentery and parietal peritoneum. Quantitative morphometric evaluation of omentum and mesentery showed a clear trend toward less angiogenesis after treatment with the AA-PDF compared to the glucose-containing PDF, which reached statistical significance in the parietal peritoneum (p < 0.04). Instillation of AA-PDF resulted in approximately 50% reduction of fibrosis in the mesentery (p < 0.04) and approximately 25% reduction in the parietal peritoneum (p < 0.009) compared to glucose-containing PDF. Glucose-containing PDF damaged the mesothelial cell layer, whereas the mesotheium was intact after AA-PDF treatment, as evidenced by electron microscopy. CONCLUSION: Our data in a rat chronic peritoneal exposure model clearly demonstrate reduced immune activation (evidenced by decreased number of rolling leukocytes and decreased induction of omental milky spots) and reduced neoangiogenesis, fibrosis, and mesothelial damage of the peritoneal membrane after treatment with AA-PDF compared to glucose-containing PDF