Screening method and therapy with agonists of DDAH I
An agonist and pharmaceutical technology, which can be used in the screening of DDAH I agonists and the field of therapy with DDAH I agonists, which can solve problems such as harm to liver function and low hepatic blood flow.
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Embodiment 1
[0103] Example 1: Infliximab Treatment Increases DDAH1 Levels and Decreases Portal Pressure
[0104] Three groups of rats were used in these experiments: BDL rats treated with vehicle, BDL rats treated with the anti-TNF monoclonal antibody infliximab, and mock-treated rats.
[0105] Such as figure 1 As shown, bile duct ligation was found to significantly reduce eNOS activity, but treatment of BDL rats with infliximab restored eNOS activity to levels similar to those observed in mock animals.
[0106] Such as figure 2 As shown, bile duct ligation also resulted in significantly higher ADMA concentrations in liver tissue compared to mock animals. However, treatment with infliximab significantly reduced ADMA levels.
[0107] Such as image 3 As shown, bile duct ligation significantly reduced the expression of DDAH-1 isoforms in the liver. After treatment with infliximab, the expression level of DDAH-1 was restored to the level of mock animals.
[0108] Such as Figure 4 ...
Embodiment 2
[0114] Example 2: The farnesoid receptor agonist INT-747 increases DDAH1 levels and decreases gating Venous pressure
[0115] Farnesoid receptor (FXR) is a bile acid responsive nuclear receptor previously shown to be hepatoprotective against bile duct ligation (BDL) injury in rats. FXR agonists have a variety of target genes including DDAH1. This study tested the hypothesis that an FXR agonist (INT-747) would restore DDAH-1 levels thereby restoring eNOS activity and reducing portal pressure.
[0116] Four weeks after BDL or mock surgery in Sprague-Dawley rats (n=14), BDL rats were gavaged with 5 mg / kg of the FXR agonist INT-747 (6-ethylchenodeoxy) in vehicle (corn oil) Cholic acid, Intercept Pharmaceuticals Inc.) for 5 days or gavage vehicle only.
[0117] Rats were evaluated for direct portal pressure after 5 days of treatment and sacrificed, and plasma and liver tissue were collected for analysis. eNOS activity was radiometrically determined by the conversion of label...
Embodiment 3
[0123] It has been demonstrated that injection of naked plasmid DNA with a promoter effective in mammalian cells results in efficient hepatic transduction of the gene of interest in rodents (Maruyama et al. J Gene Med 2002, 4:333-41). This approach, termed 'hydrodynamic gene therapy', was used to determine the effect of DDAH-1 transduction on portal pressure in BDL rats.
[0124] Plasmids containing human DDAH-1 cDNA were injected into BDL rodents or mock rodents generated as above via a branch of the jugular vein. A control plasmid containing GFP was used as a control for intervention in BDL animals. Animals underwent direct portal pressure measurements 48 hours post-intervention, and plasma and tissue were collected for analysis of hepatic DDAH-1 mRNA (rtPCR) and protein expression (Western blot), as well as routine histology and biochemistry.
[0125] All animals treated tolerated hydrodynamic injections well and were provided ad libitum access to food and water. 48 hours...
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