Hypertension-Associated Genes in the Mesenteric Artery of Three Spontaneously Hypertensive Rat Substrains Identified Using a DNA Array Method.

BACKGROUND: Although the mesenteric artery plays a key role in regulating peripheral blood pressure, the molecular mechanisms that underlie the development of essential hypertension are not yet fully understood. MATERIALS AND METHODS: We explored candidate genes for hypertension using three related strains of spontaneously hypertensive rats (SHRs) that mimic human essential hypertension. In this study we used DNA microarrays, a powerful tool for studying genetic diseases, to compare gene expression in the mesenteric artery of three SHR substrains: SHR, stroke-prone SHR (SHRSP), and malignant SHRSP (M-SHRSP). RESULTS: Compared to normotensive 6-week old Wistar Kyoto rats (WKY), higher blood pressure correlated with overexpression of 31 genes and with down regulation of 24 genes. Adam23, which negatively regulates potassium current, and the potassium channel genes, Kcnc2 and Kcnq5, were associated with the onset of hypertension. In addition, Spock2 and Agtrap were identified as strengtheners of hypertension by analyzing up and down regulated genes at 9-weeks of age. CONCLUSIONS: Adam23, Kcnc2 and Kcnq5 appear to be factors for the onset of hypertension, while Spock2 and Agtrap are as factors that strengthen hypertension. These findings contribute to our understanding of the pathophysiology of hypertension and to the development of treatment for this condition.

Thromboxane A2 receptor antagonist (ONO-8809) attenuates renal disorders caused by salt overload in stroke-prone spontaneously hypertensive rats.

Epidemiological and clinical studies have demonstrated that excessive salt intake causes severe hypertension and exacerbates organ derangement, such as in chronic kidney disease (CKD). In this study, we focused on evaluating the histological and gene expression effects in the kidneys of stroke-prone spontaneously hypertensive rats (SHRSP) with a high salt intake and the thromboxane A2 / prostaglandin H2 receptor (TPR) blocker ONO-8809. Six-week-old SHRSPs were divided into three groups and were fed normal chow containing 0.4% NaCl, 2.0%NaCl or 2.0%NaCl + ONO-8809 (0.6 mg/kg p.o. daily). Histological analyses with immunohistochemistry and a gene expression assay with a DNA kidney microarray were performed after 8 weeks. The following changes were observed in SHRSPs with the high salt intake. Glomerular sclerotic changes were remarkably observed in the juxtamedullary cortex areas. The ED1, monocyte chemoattractant protein-1 (MCP-1), nitrotyrosine and hypoxia inducible factor 1α (HIF-1α) staining areas were increased in the glomeruli and interstitial portion of the kidneys. The genes Tbxa2r (that encodes TPR), Prcp and Car7 were significantly underexpressed in the kidneys. The plasma 8-isoprostane level was significantly elevated and was attenuated with the ONO-8809 treatment. Thromboxane A2 (TXA2 ) and oxidative stress exaggerated renal dysfunction in the salt-loaded SHRSPs, and ONO-8809 as a TPR blocker suppressed these changes. Therefore, ONO-8809 is a candidate drug to prevent CKD in hypertensive patients when CKD is associated with a high salt intake.

Whole rat DNA array survey for candidate genes related to hypertension in kidneys from three spontaneously hypertensive rat substrains at two stages of age and with hypotensive induction caused by hydralazine hydrochloride.

Clarification of the genetic nature and more effective care for hypertension are required, given the high incidences of cardiovascular and cerebrovascular mortality. Thus, we surveyed candidate genes for hypertension with rat whole gene DNA microarrays using three novel methods. Gene expression analyses were conducted as follows: Method 1, three types of spontaneously hypertensive rat (SHR) substrains, SHR, stroke-prone SHR (SHRSP) and malignant type of SHRSP (M-SHRSP) were used and compared to normotensive Wistar Kyoto rats; Method 2, the expressed genes between rats of different ages were compared for different blood pressures; and Method 3, genes that were expressed in rats treated with or without an acute hypotensive stimulus, the antihypertensive hydralazine hydrochloride, were compared. This approach identified dozens of genes, including Dusp15, Cyp8b1, Armc 3, Gtpbp4, Mettl2, Mapk14, Prkar2b, frame 12, Anxa13, Ephx2, Myr8 and Pcdh9 by Method 1; Cyp2C and Atp12a by Method 2; and Kcnc3, Vnn1, TC560558 and Gabrq and a number of unknown genes by Methods 2 and 3, as probable candidate genes for hypertension in SHR substrains. Ephx2 was previously reported as a candidate gene in SHRs; however other genes were identified for the first time in this study. Since it was not always possible to completely demonstrate that these genes are responsible for hypertension in SHRs, further research into true candidate genes that participate in the genesis of hypertension in SHR substrains is warranted.

Gene expression in the adrenal glands of three spontaneously hypertensive rat substrains.

We examined gene expression profiles in rat adrenal glands using genome-wide microarray technology. Gene expression levels were determined in four rat strains, including one normotensive strain [Wistar-Kyoto (WKY)] and three substrains derived from WKY rats: spontaneously hypertensive rats (SHR), stroke-prone SHR (SHRSP) and malignant SHRSP (M-SHRSP). This study represents the first attempt at using microarrays to compare gene expression profiles in SHR, SHRSP and M-SHRSP adrenal glands, employing WKY as controls. Expression measurements were made in these four rat strains at 6 and 9 weeks of age; 6 weeks of age covers the pre-hypertensive period in SHR and SHRSP, and 9 weeks of age is the period of rapidly rising blood pressure (BP). Since the aim of this study was to identify candidate genes involved in the genesis of hypertension in the SHR substrains, we identified genes that were consistently different in their expression, isolating 87 up-regulated genes showing a more than 4-fold increase and 128 down-regulated genes showing a less than 1/4-fold decrease in at least two different experiments. We classified all these up- or down-regulated genes by their expression profiles, and searched for candidate genes. At 6 weeks of age, several BP-regulating genes including sparc/osteonectin (Spock2), kynureninase (Kynu), regulator of G-protein signaling 2 (Rgs2) and gap junction protein α1 (Gja1) were identified as up-regulated, and urotensin 2 (Uts2), cytoplasmic epoxide hydrolase 2 (Ephx2), apelin (Apln), insulin-like growth factor 1 receptor (Igf1r) and angiotensin II receptor-associated protein (Agtrap) were identified as down-regulated. The Kynu and Ephx2 genes have previously been reported by other groups to be responsible for hypertension in SHR; however, our present approach identified at least seven new candidate genes.