Gel filtration chromatography was utilized for further purification

n rate and diuresis, which is reversed by co-administration of the NO precursor L-arginine, indicating the role of this pathway in vascular and renal pathophysiology. In addition, ADMA is elevated in patients with HTN and IR and is implicated in the progression of salt-sensitive HTN. In a preclinical study, overexpression of DDAH is associated with enhanced insulin sensitivity. In addition, pharmacological manipulation of DDAH using FXR agonists is known to reduce ADMA and improve insulin sensitivity in humans suggesting that DDAH may 18362028 be an attractive target to improve insulin sensitivity. In fact, we have been carrying out efforts to discover and validate agents that enhance DDAH activity using the method we described. INT-747 does not favorably influence tissue DDAH activity/circulating ADMA/NO levels, nor reverse saltsensitive HTN. In the present study, we evaluated the Discussion Novelty and Significance: The simultaneous effect of INT-747, a small molecule FXR agonist, on blood pressure and insulin sensitivity has not been studied before in Dahl Rats; an animal model that displays both systemic hypertension and IR. Our study addressed the scientific curiosity of whether it is possible to concurrently modulate both BP and insulin sensitivity using INT-747. We have also addressed the effect of high-salt diet on DDAH expression. Our key findings are: 1) INT-747 does not reduce systemic or pulmonary vascular pressure in high-salt fed Dahl rats. 2) INT-747 induces hepatic DDAH expression and enhances insulin sensitivity in this animal model as described below. Role of DDAH/ADMA/NO in salt-sensitive hypertension and insulin resistance. There is mounting pharmacological and Upregulation of DDAH and Insulin Sensitivity 747 ameliorates nephropathy in animal models of diabetes, associated with reductions in hyperlipoproteinemia, fibrosis, proteinuria, inflammation, and oxidative stress. Moreover, in 5/6 nephrectomized ApoE-deficient mice, INT-747 reduces PD-1/PD-L1 inhibitor 2 chronic kidney disease -induced vascular calcification independent of atherosclerosis progression. However, our study of renal histology showed comparable TMA between the INT-747 and vehicle treated groups. INT-747 favorably effects hepatic DDAH expression but not activity. More intriguingly, we demonstrate that HS-diet significantly downregulated hepatic DDAH1 expression. However, treatment with INT-747 protected the loss in hepatic DDAH1. Our observations are similar to those in an animal model of bile duct ligation injury where treatment with INT747 increases hepatic DDAH1 expression and improves portal pressure. Indeed, a small clinical study evaluating the therapeutic potential of INT-747 in regulating portal hypertension in patients with alcoholic cirrhosis has just completed. Although INT747 induced hepatic DDAH expression likely due to the presence of a putative FXR response element in the DDAH1 promoter, 11121575 it did not favorably influence DDAH activity, nor did it favorably influence circulating ADMA and NO levels in this model. INT-747 improves insulin sensitivity. Salt loading aggravates insulin resistance in Dahl rats; an animal model that inherently develops IR at weaning and prior to salt-loading. INT-747 enhanced insulin sensitivity in these animals as demonstrated by the reduction in IR index. It is possible that this effect is mediated in part by an increase in hepatic DDAH protein expression. However, the improvement in insulin sensitivity may be independent of any effect of IN