Recent data from animal models indicate that the eNOS null mice present a phenotype that resemble the human metabolic syndrome (hypertension, insulin resistance and hypertriglyceridemia). In this work, we have studied whether NOS3 gene, previously related to endothelial dysfunction, might have a role in metabolic syndrome susceptibility in hypertensive patients. To carry out the study, we genotyped 105 hypertensive patients < or = 60 years old with two polymorphisms of NOS3 gene: 1132 T>C and 7164 G>T (GeneBank:AF519768.1). To check the allelic frequency of these polymorphisms in our geographical area, we also genotyped 94 unselected healthy controls (control group). To perform sample genotyping, we designed a novel FRET system coupled to real time PCR. There were no differences in genotypic distribution or allelic frequency between hypertensive patients and the control group. However, we observed that 786CC genotype was significantly more frequent in hypertensive patients with metabolic syndrome than in those without the syndrome (p=0.0022). When both polymorphisms were analyzed, we identified the 786C894G as the risk haplotype for metabolic syndrome susceptibility (p=0.011). These data suggest a role of the NOS3 gene in the pathogenesis of metabolic syndrome in hypertensive patients.

Recently, we reported that the polymorphism 1132T>C (GenBank: AF519768.1) of the NOS3 gene was associated with susceptibility to metabolic syndrome (MS) in hypertensive patients. This suggests that other genes such as CAV1, whose product (CAV1) regulates eNOS activity, could also be related to this phenotype. In this work we investigated the following: i) whether CAV1 is a quantitative trait locus of clustering of atherothrombotic traits associated with MS; ii) whether CVA1 is associated with hypertension or MS in hypertensive patients; and iii) whether genetic interaction between NOS3 and CAV1 is involved in the susceptibility or protection to hypertension associated with MS. To carry out the study, we genotyped 285 randomly selected individuals and 175 hypertensive patients, all of them < or = 60 years old, with two polymorphisms of the CAV1 gene: the 22285 C>T and the 22375-22375 del AC (GenBank AF125348), and the 1132T>C polymorphism of the NOS3 gene. To perform sample genotyping, we used pyrosequencing and FRET techniques. The 22285 C-22375-22375 del (Cd) haplotype of CAV1 gene was associated with low levels of blood pressure in the general population. Moreover, it was a genetic protection factor against MS in hypertensive patients. In addition, we found no evidence of gene-gene interaction between NOS3 and CAV1 genes with regard to that phenotype.

Phytochemical and biological studies aimed at the discovery and development of novel anti-inflammatory agents from natural sources have been conducted in our laboratories for a number of years. In the course of our investigations, we have been attempting to isolate the anti-inflammatory principles from a number of botanical species, some of which are used for the treatment of inflammatory diseases in Spanish traditional medicine. Jasonia glutinosa D.C. (Chiliadenus saxatilis CASS.ϭChiliadenus glutinosus (L.) FOURR.) is a member of the subtribe Inulinae, tribe Inuleae, Fam. Asteraceae and occurs in the Mediterranean and adjacent areas. [1][2][3][4] In both folk and modern traditional medicine, this plant, under the name of "te de roca," is very popular as a stomachic as well as a component in preparations showing beneficial effects such as antispasmodic and as an anti-inflammatory drug. The tea made from its dried leaves is a popular beverage in the Iberian Peninsula. 5) However, no reports as to its experimental biological activity have been found in the literature.In a previous study performed as part of our collaborative search for novel anti-inflammatory agents from plants, the leaves of J. glutinosa were found to contain flavonol glycosides and four sesquiterpenes: lucinone, glutinone, 5-epi-kutdtriol and kutdtriol. 6-8) Plant sesquiterpenes are secondary metabolites widely distributed in the higher plant kingdom and are known to show diverse biological and pharmacological actions, including anti-inflammatory activity. [9][10][11] Since the effect of sesquiterpenes found in the leaves of J. glutinosa on the key enzymes of inflammation has not yet been studied, their inhibitory activities on cyclooxygenase-1 (COX-1) (PGsynthase and TX-synthase) and 5-lipoxygenase (5-LOX) have been investigated in the present paper. MATERIALS AND METHODS Test CompoundsLucinone, glutinone, 5-epi-kutdtriol and kutdtriol were isolated from J. glutinosa as described previously by us 6-8) (Fig. 1). All compounds were dissolved in dimethylsulfoxide (DMSO) to an appropriate concentration and assayed for inhibition of prostaglandin E 2 (PGE 2 ) and leukotriene C 4 (LTC 4 ) from mouse peritoneal cells and thromboxane B 2 (TXB 2 ) from human platelets. Indomethacin (25-100 mM), nordihydroguaiaretic acid (NDGA) (6.25-25 mM) and ibuprofen (25-100 mM) were used as reference compounds (PGE 2 , LTC 4 and TXB 2 , respectively). All chemicals used were purchased from Sigma Chemical Co. (U.S.A.).Cell Viability Cell viability was assessed by using a 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-based colorimetric assay. 12) Cells in 96-well plates (8ϫ10 5 cells/well) were exposed to various concentrations of samples at 37°C, 5% CO 2 in air for 3 h. The 20 ml MTT solution (5 mg/ml in phosphate buffered saline) was added and further incubated for 3 h at 37°C. After aspirating the supernatant from the wells, 100 ml of DMSO were added for the dissolution of formazan crystals. The absorbance of each well was then read at 520 nm ...

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