The tissue comprised both glomerular and tubulo-interstitial elements. Given that the tubulointerstitium occupies up to 90 of the total kidney volume, any changes in collagen type III and fibronectin transcripts in the glomerular compartment following sulodexide treatment may beSulodexide and Diabetic Nephropathymasked by its effect on the tubulo-interstitium. Since TGF-b1 expression is reduced in DN mice following sulodexide treatment, it is likely that sulodexide-mediated increase in collagen type III and fibronectin expression is through a mechanism that is independent of TGF-b1. Rossini et al demonstrated that sulodexide could ameliorate early but not late stages of kidney disease in a murine model of type II DN [46], but in contrast to our 38916-34-6 web studies, these researchers did not report any induction of matrix protein synthesis by sulodexide. This anomaly may be due to different pathogenic mechanisms induced in type I and II DN mouse models and method of sulodexide administration. In a mild nonhypertensive rat model of chronic kidney disease, sulodexide improved renal function, although the beneficial effects of this drug was not sustained [46], an observation that was also observed in our study, whereby serum creatinine levels were reduced after 8 weeks treatment, but subsequently had no effect at later timepoints, possibly due to alterations in the structural integrity of the glomerulus following drug treatment. Although all resident renal cells participate in renal fibrosis, the accumulation of matrix proteins within the glomerulus during pathological conditions is initiated in the mesangium. Mesangial cells were therefore utilized to investigate the effect of sulodexide on matrix protein synthesis in vitro. We demonstrated that both PKC and ERK signaling pathways regulated the synthesis of matrix proteins in mesangial cells and reduced I-BRD9 custom synthesis phosphorylation of PKC isomers and ERK significantly decreased fibronectin and collagen type III synthesis. Under our experimental setting, MMC constitutively expressed phosphorylated ERK, PKC-a and PKCbII but not PKC-bI. Elevated glucose concentrations was shown to increase ERK, PKC-a and PKC-bII phosphorylation and induce PKC-bI activation in MMC. The effect of sulodexide on PKC and ERK signaling pathways under physiological and experimental conditions was selective, whereby sulodexide markedly attenuated ERK and PKC-bII phosphorylation in control and 30 mM D-glucose stimulated cells, but had no effect on PKC-a or PKC-bI. These results corroborate our in vivo findings. The role of PKC-bI in mediating fibrotic processes in the kidney is well established [47?9]. Increased collagen type III and fibronectin synthesis in MMC was observed following their exposure to sulodexide, and their synthesis was further exacerbated by sulodexide in the presence of elevated glucose concentration. Based on these findings, it is plausible to suggest that the observed increase in fibronectin and collagen 16402044 type III expression in the glomeruli of DN mice was directly attributed to the effect of sulodexide on mesangial cells. A schematic diagram summarizing our in vivo and in vitro data is shown in Figure 14. In conclusion, we have demonstrated that sulodexide treatment reduced albuminuria, improved serum levels of urea, restored perlecan expression and ameliorated selective renal histopathologic changes in male C57BL/6 DN mice that included reduced collagen type I and IV deposition, and ERK and PKC-bII activation. In contr.The tissue comprised both glomerular and tubulo-interstitial elements. Given that the tubulointerstitium occupies up to 90 of the total kidney volume, any changes in collagen type III and fibronectin transcripts in the glomerular compartment following sulodexide treatment may beSulodexide and Diabetic Nephropathymasked by its effect on the tubulo-interstitium. Since TGF-b1 expression is reduced in DN mice following sulodexide treatment, it is likely that sulodexide-mediated increase in collagen type III and fibronectin expression is through a mechanism that is independent of TGF-b1. Rossini et al demonstrated that sulodexide could ameliorate early but not late stages of kidney disease in a murine model of type II DN [46], but in contrast to our studies, these researchers did not report any induction of matrix protein synthesis by sulodexide. This anomaly may be due to different pathogenic mechanisms induced in type I and II DN mouse models and method of sulodexide administration. In a mild nonhypertensive rat model of chronic kidney disease, sulodexide improved renal function, although the beneficial effects of this drug was not sustained [46], an observation that was also observed in our study, whereby serum creatinine levels were reduced after 8 weeks treatment, but subsequently had no effect at later timepoints, possibly due to alterations in the structural integrity of the glomerulus following drug treatment. Although all resident renal cells participate in renal fibrosis, the accumulation of matrix proteins within the glomerulus during pathological conditions is initiated in the mesangium. Mesangial cells were therefore utilized to investigate the effect of sulodexide on matrix protein synthesis in vitro. We demonstrated that both PKC and ERK signaling pathways regulated the synthesis of matrix proteins in mesangial cells and reduced phosphorylation of PKC isomers and ERK significantly decreased fibronectin and collagen type III synthesis. Under our experimental setting, MMC constitutively expressed phosphorylated ERK, PKC-a and PKCbII but not PKC-bI. Elevated glucose concentrations was shown to increase ERK, PKC-a and PKC-bII phosphorylation and induce PKC-bI activation in MMC. The effect of sulodexide on PKC and ERK signaling pathways under physiological and experimental conditions was selective, whereby sulodexide markedly attenuated ERK and PKC-bII phosphorylation in control and 30 mM D-glucose stimulated cells, but had no effect on PKC-a or PKC-bI. These results corroborate our in vivo findings. The role of PKC-bI in mediating fibrotic processes in the kidney is well established [47?9]. Increased collagen type III and fibronectin synthesis in MMC was observed following their exposure to sulodexide, and their synthesis was further exacerbated by sulodexide in the presence of elevated glucose concentration. Based on these findings, it is plausible to suggest that the observed increase in fibronectin and collagen 16402044 type III expression in the glomeruli of DN mice was directly attributed to the effect of sulodexide on mesangial cells. A schematic diagram summarizing our in vivo and in vitro data is shown in Figure 14. In conclusion, we have demonstrated that sulodexide treatment reduced albuminuria, improved serum levels of urea, restored perlecan expression and ameliorated selective renal histopathologic changes in male C57BL/6 DN mice that included reduced collagen type I and IV deposition, and ERK and PKC-bII activation. In contr.
