Sp1 decoy oligodeoxynucleotide decreases angiotensin receptor expression and blood pressure in spontaneously hypertensive rats

Brain Res. 2003 Nov 28;992(1):1-8. doi: 10.1016/s0006-8993(03)03534-0.

Abstract

The transcriptional factor Sp1 is associated with GC-rich promoters and involved in basal promoter activity. A GC-box-related sequence is located within the -58 to -34 base pair region of the angiotensin type 1 receptor gene promoter. We examined whether Sp1 in the hypothalamus was increased in spontaneously hypertensive rats (SHR) and whether inhibition of Sp1 binding sites suppressed angiotensin type 1 receptor expression and thus decreased blood pressure in SHR. Western blot analysis showed that Sp1 protein levels were increased in nuclear extracts of hypothalamus from SHR. Electrophoretic mobility shift assay (EMSA) using oligonucleotides containing Sp1 consensus sequence and -58 to -34 region sequence oligonucleotides showed that DNA-protein complexes were greater in nuclear extracts of hypothalamus from SHR than those of Wistar Kyoto rats (WKY). Sp1 decoy phosphorothioate oligodeoxynucleotides injected into the lateral ventricle produced a decrease in blood pressure in SHR, and decreased angiotensin type 1 receptor mRNA levels and number of angiotensin receptors in the hypothalamus of SHR. Pressor responses to angiotensin II but not to carbachol injected into the lateral ventricle were decreased in the Sp1 decoy-treated SHR. The results of the present study suggest that Sp1 levels in the hypothalamus of SHR are increased, and that inhibition of the binding of Sp1 to its binding sites decreases angiotensin type 1 receptor expression and blood pressure in SHR. The possibility cannot be ruled out that the Sp1 decoy oligodeoxynucleotides (ODN) also suppressed transcriptions of genes other than the angiotensin type 1 receptor gene.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Pressure / drug effects
  • Blood Pressure / physiology*
  • Blotting, Western
  • Electrophoretic Mobility Shift Assay
  • Hypothalamus / metabolism*
  • Injections, Intraventricular
  • Male
  • Oligonucleotides, Antisense / administration & dosage
  • Oligonucleotides, Antisense / pharmacology*
  • RNA, Messenger / analysis
  • Rats
  • Rats, Inbred SHR
  • Rats, Inbred WKY
  • Receptors, Angiotensin / biosynthesis*
  • Receptors, Angiotensin / drug effects
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sp1 Transcription Factor / genetics
  • Sp1 Transcription Factor / metabolism*

Substances

  • Oligonucleotides, Antisense
  • RNA, Messenger
  • Receptors, Angiotensin
  • Sp1 Transcription Factor