17β-Estradiol inhibits subarachnoid hemorrhage-induced inducible nitric oxide synthase gene expression by interfering with the nuclear factor κB transactivation

Huei Chuan Shih, Chih Lung Lin, Tzu Ying Lee, Wen Sen Lee, Chin Hsu

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

BACKGROUND AND PURPOSE - Previously, we showed that 17β-estradiol (E2) treatment prevented the subarachnoid hemorrhage (SAH)-induced cerebral vasospasm in male rats. The aim of this study was designed to further delineate the molecular mechanisms underlying E2-induced inhibition of inducible nitric oxide synthase (iNOS) upregulation and relief of vasospasm caused by SAH. METHODS - The 2-hemorrhage SAH model was induced by 2 autologous injections of blood into the cisterna magna of adult male rats. The rats were then subcutaneously implanted of a Silastic tube containing corn oil with or without 17β-estradiol benzoate and received daily intraperitoneal injections of various doses of ICI 182,780, a nonselective estrogen receptor (ER) antagonist, for 7 days after the first hemorrhage. Basilar arteries were then removed for protein extraction, RNA isolation, and gel mobility assay. The protein levels of iNOS, p65, and ER were examined by Western blot analysis, and that iNOS mRNA expression was evaluated by reverse-transcription polymerase chain reaction. RESULTS - E2 prevented the SAH-induced vasospasm and increases of the levels of iNOS protein and mRNA in basilar artery through an ER-dependent mechanism. Treatment of the SAH rat with E2 did not affect the nuclear translocation of p65 subunit of nuclear factor κB, but caused an increase of the association of p65/ER, and reversed the SAH-induced increase of the p65 binding on iNOS promoter. CONCLUSIONS - E2 inhibits the SAH-induced increase of iNOS by increasing the association of p65/ER, which in turn inhibits the binding of p65 to iNOS DNA. Our data suggest the potential applications of E2 in the treatment of SAH patient.

Original languageEnglish
Pages (from-to)3025-3031
Number of pages7
JournalStroke
Volume37
Issue number12
DOIs
Publication statusPublished - Dec 2006

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Nitric Oxide Synthase Type II
Subarachnoid Hemorrhage
Transcriptional Activation
Estradiol
Gene Expression
Estrogen Receptors
Basilar Artery
Hemorrhage
Cisterna Magna
Intracranial Vasospasm
Messenger RNA
Proteins
Corn Oil
Intraperitoneal Injections
Reverse Transcription
Up-Regulation
Therapeutics
Western Blotting
Gels
RNA

Keywords

  • Estradiol
  • iNOS
  • Nuclear factor κB
  • Stroke
  • Subarachnoid hemorrhage

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Neuroscience(all)
  • Medicine(all)

Cite this

17β-Estradiol inhibits subarachnoid hemorrhage-induced inducible nitric oxide synthase gene expression by interfering with the nuclear factor κB transactivation. / Shih, Huei Chuan; Lin, Chih Lung; Lee, Tzu Ying; Lee, Wen Sen; Hsu, Chin.

In: Stroke, Vol. 37, No. 12, 12.2006, p. 3025-3031.

Research output: Contribution to journalArticle

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abstract = "BACKGROUND AND PURPOSE - Previously, we showed that 17β-estradiol (E2) treatment prevented the subarachnoid hemorrhage (SAH)-induced cerebral vasospasm in male rats. The aim of this study was designed to further delineate the molecular mechanisms underlying E2-induced inhibition of inducible nitric oxide synthase (iNOS) upregulation and relief of vasospasm caused by SAH. METHODS - The 2-hemorrhage SAH model was induced by 2 autologous injections of blood into the cisterna magna of adult male rats. The rats were then subcutaneously implanted of a Silastic tube containing corn oil with or without 17β-estradiol benzoate and received daily intraperitoneal injections of various doses of ICI 182,780, a nonselective estrogen receptor (ER) antagonist, for 7 days after the first hemorrhage. Basilar arteries were then removed for protein extraction, RNA isolation, and gel mobility assay. The protein levels of iNOS, p65, and ER were examined by Western blot analysis, and that iNOS mRNA expression was evaluated by reverse-transcription polymerase chain reaction. RESULTS - E2 prevented the SAH-induced vasospasm and increases of the levels of iNOS protein and mRNA in basilar artery through an ER-dependent mechanism. Treatment of the SAH rat with E2 did not affect the nuclear translocation of p65 subunit of nuclear factor κB, but caused an increase of the association of p65/ER, and reversed the SAH-induced increase of the p65 binding on iNOS promoter. CONCLUSIONS - E2 inhibits the SAH-induced increase of iNOS by increasing the association of p65/ER, which in turn inhibits the binding of p65 to iNOS DNA. Our data suggest the potential applications of E2 in the treatment of SAH patient.",
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T1 - 17β-Estradiol inhibits subarachnoid hemorrhage-induced inducible nitric oxide synthase gene expression by interfering with the nuclear factor κB transactivation

AU - Shih, Huei Chuan

AU - Lin, Chih Lung

AU - Lee, Tzu Ying

AU - Lee, Wen Sen

AU - Hsu, Chin

PY - 2006/12

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N2 - BACKGROUND AND PURPOSE - Previously, we showed that 17β-estradiol (E2) treatment prevented the subarachnoid hemorrhage (SAH)-induced cerebral vasospasm in male rats. The aim of this study was designed to further delineate the molecular mechanisms underlying E2-induced inhibition of inducible nitric oxide synthase (iNOS) upregulation and relief of vasospasm caused by SAH. METHODS - The 2-hemorrhage SAH model was induced by 2 autologous injections of blood into the cisterna magna of adult male rats. The rats were then subcutaneously implanted of a Silastic tube containing corn oil with or without 17β-estradiol benzoate and received daily intraperitoneal injections of various doses of ICI 182,780, a nonselective estrogen receptor (ER) antagonist, for 7 days after the first hemorrhage. Basilar arteries were then removed for protein extraction, RNA isolation, and gel mobility assay. The protein levels of iNOS, p65, and ER were examined by Western blot analysis, and that iNOS mRNA expression was evaluated by reverse-transcription polymerase chain reaction. RESULTS - E2 prevented the SAH-induced vasospasm and increases of the levels of iNOS protein and mRNA in basilar artery through an ER-dependent mechanism. Treatment of the SAH rat with E2 did not affect the nuclear translocation of p65 subunit of nuclear factor κB, but caused an increase of the association of p65/ER, and reversed the SAH-induced increase of the p65 binding on iNOS promoter. CONCLUSIONS - E2 inhibits the SAH-induced increase of iNOS by increasing the association of p65/ER, which in turn inhibits the binding of p65 to iNOS DNA. Our data suggest the potential applications of E2 in the treatment of SAH patient.

AB - BACKGROUND AND PURPOSE - Previously, we showed that 17β-estradiol (E2) treatment prevented the subarachnoid hemorrhage (SAH)-induced cerebral vasospasm in male rats. The aim of this study was designed to further delineate the molecular mechanisms underlying E2-induced inhibition of inducible nitric oxide synthase (iNOS) upregulation and relief of vasospasm caused by SAH. METHODS - The 2-hemorrhage SAH model was induced by 2 autologous injections of blood into the cisterna magna of adult male rats. The rats were then subcutaneously implanted of a Silastic tube containing corn oil with or without 17β-estradiol benzoate and received daily intraperitoneal injections of various doses of ICI 182,780, a nonselective estrogen receptor (ER) antagonist, for 7 days after the first hemorrhage. Basilar arteries were then removed for protein extraction, RNA isolation, and gel mobility assay. The protein levels of iNOS, p65, and ER were examined by Western blot analysis, and that iNOS mRNA expression was evaluated by reverse-transcription polymerase chain reaction. RESULTS - E2 prevented the SAH-induced vasospasm and increases of the levels of iNOS protein and mRNA in basilar artery through an ER-dependent mechanism. Treatment of the SAH rat with E2 did not affect the nuclear translocation of p65 subunit of nuclear factor κB, but caused an increase of the association of p65/ER, and reversed the SAH-induced increase of the p65 binding on iNOS promoter. CONCLUSIONS - E2 inhibits the SAH-induced increase of iNOS by increasing the association of p65/ER, which in turn inhibits the binding of p65 to iNOS DNA. Our data suggest the potential applications of E2 in the treatment of SAH patient.

KW - Estradiol

KW - iNOS

KW - Nuclear factor κB

KW - Stroke

KW - Subarachnoid hemorrhage

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