Attenuation of subarachnoid hemorrhage–induced apoptotic cell death with 17 beta-estradiol

Laboratory investigation

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Object

Apoptosis is implicated in vasospasm and long-term sequelae of subarachnoid hemorrhage (SAH). The authors observed that 17β-estradiol (E2) can attenuate cerebral vasospasm, lower endothelin-1 production, and preserve normal endothelial nitric oxide synthase expression by reduction of inducible NO synthase expression in experimental SAH. The authors investigated the potential antiapoptotic effects of E2 in an experimental rat model of SAH.

Methods

The authors examined the antiapoptotic effects of E2 in a double-hemorrhage SAH model in male Sprague-Dawley rats. The rats underwent subcutaneous implantation of a Silastic tube containing corn oil either with or without E2, and some E2-treated animals also received ICI 182,780 (a nonselective estrogen receptor [ER] antagonist) for 7 days after SAH. The degree of vasospasm was determined by averaging the cross-sectional areas of the basilar artery 7 days after SAH. The expression of apoptotic indicators, including TNF-α, caspase 3, Bcl-2, Bax, terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling (TUNEL), and cell death assays were used for detection of apoptosis.

Results

Treatment with E2 significantly attenuated SAH-induced vasospasm. Seven days after the induction of SAH, positive TUNEL-staining was seen, and DNA fragmentation was increased in the dentate gyrus. Increased TNF-α and cleaved caspase-3 protein expression and decreased Bcl-2 protein expression in the dentate gyrus were also observed. These changes were reversed with E2-treatment but not in the presence of ICI 182,780. However, the expression of Bax did not change after SAH either with or without E2 treatment.

Conclusions

The authors found that E2 appears to confer an antiapoptotic effect that reduces secondary brain injury after SAH via estrogen receptor–dependent mechanisms. This finding provides support for possible future applications of E2 treatment for the reduction of secondary injury after SAH in patients.

Abbreviations used in this paper: BA = basilar artery; ELISA = enzyme-linked immunosorbent assay; eNOS = endothelial nitric oxide synthase; ER = estrogen receptor; E2 = 17β-estradiol; iNOS = inducible NOS; SAH = subarachnoid hemorrhage; TUNEL = terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling.
Article Information

Contributor Notes

Address correspondence to: Shiuh-Lin Hwang, M.D., Ph.D., Division of Neurosurgery, Kaohsiung Medical University Hospital, No. 100, Shih-Chuan 1st Road, Kaohsiung 807, Taiwan. email: nsdoctor@yam.com.Please include this information when citing this paper: published online May 8, 2009; DOI: 10.3171/2009.3.JNS081660.
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References
  • 1

    Amantea DRusso RBagetta GCorasaniti MT: From clinical evidence to molecular mechanisms underlying neuroprotection afforded by estrogens. Pharmacol Res 52:1191322005

    • Search Google Scholar
    • Export Citation
  • 2

    Buchanan KMElias LJGoplen GB: Differing perspectives on outcome after subarachnoid hemorrhage: the patient, the relative, the neurosurgeon. Neurosurgery 46:8318402000

    • Search Google Scholar
    • Export Citation
  • 3

    Cahill JCalvert JWZhang JH: Mechanisms of early brain injury after subarachnoid hemorrhage. J Cereb Blood Flow Metab 26:134113532006

    • Search Google Scholar
    • Export Citation
  • 4

    Conrad CDRoy EJ: Dentate gyrus destruction and spatial learning impairment after corticosteroid removal in young and middle-aged rats. Hippocampus 5:1151995

    • Search Google Scholar
    • Export Citation
  • 5

    Culmsee CVedder HRavati AJunker VOtto DAhlemeyer B: Neuroprotection by estrogens in a mouse model of focal cerebral ischemia and in cultured neurons: evidence for a receptor-independent antioxidative mechanism. J Cereb Blood Flow Metab 19:126312691999

    • Search Google Scholar
    • Export Citation
  • 6

    Dhandapani KMBrann DW: Role of astrocytes in estrogen-mediated neuroprotection. Exp Gerontol 42:70752007

  • 7

    Dubal DBKashon MLPettigrew LCRen JMFinklestein SPRau SW: Estradiol protects against ischemic injury. J Cereb Blood Flow Metab 18:125312581998

    • Search Google Scholar
    • Export Citation
  • 8

    Graham SHChen JClark RS: Bcl-2 family gene products in cerebral ischemia and traumatic brain injury. J Neurotrauma 17:8318412000

    • Search Google Scholar
    • Export Citation
  • 9

    Hara AYoshimi NMori H: Evidence for apoptosis in human intracranial aneurysms. Neurol Res 20:1271301998

  • 10

    Hicks RRSmith DHLowenstein DHSaint Marie RMcIntosh TK: Mild experimental brain injury in the rat induces cognitive deficits associated with regional neuronal loss in the hippocampus. J Neurotrauma 10:4054141993

    • Search Google Scholar
    • Export Citation
  • 11

    Hütter BOKreitschmann-Andermahr IMayfrank LRohde VSpetzger UGilsbach JM: Functional outcome after aneurysmal subarachnoid hemorrhage. Acta Neurochir Suppl 72:1571741999

    • Search Google Scholar
    • Export Citation
  • 12

    Kawas CResnick SMorrison ABrookmeyer RCorrada MZonderman A: A prospective study of estrogen replacement therapy and the risk of developing Alzheimer's disease: the Baltimore Longitudinal Study of Aging. Neurology 48:151715211997

    • Search Google Scholar
    • Export Citation
  • 13

    Kondo SHashimoto NKikuchi HHazama FNagata IKataoka H: Apoptosis of medial smooth muscle cells in the development of saccular cerebral aneurysms in rats. Stroke 29:1811891998

    • Search Google Scholar
    • Export Citation
  • 14

    Lee SYAndoh TMurphy DLChiueh CC: 17beta-estradiol activates ICI 182,780-sensitive estrogen receptors and cyclic GMP-dependent thioredoxin expression for neuroprotection. FASEB J 17:9479482003

    • Search Google Scholar
    • Export Citation
  • 15

    Levin ER: Cellular functions of plasma membrane estrogen receptors. Steroids 67:4714752002

  • 16

    Li JWu MQue LWang YXu XHu Y: 17beta-estradiol attenuates cardiac dysfunction and decreases NF-kappaB binding activity in mechanically stretched rat hearts. Steroids 73:7207262008

    • Search Google Scholar
    • Export Citation
  • 17

    Lin CLShih HCDumont ASKassell NFLieu ASSu YF: The effect of 17beta-estradiol in attenuating experimental subarachnoid hemorrhage-induced cerebral vasospasm. J Neurosurg 104:2983042006

    • Search Google Scholar
    • Export Citation
  • 18

    Liu ZGastard MVerina TBora SMouton PRKoliatsos VE: Estrogens modulate experimentally induced apoptosis of granule cells in the adult hippocampus. J Comp Neurol 441:182001

    • Search Google Scholar
    • Export Citation
  • 19

    Maggi ACiana PBelcredito SVegeto E: Estrogens in the nervous system: mechanisms and nonreproductive functions. Annu Rev Physiol 66:2913132004

    • Search Google Scholar
    • Export Citation
  • 20

    Matz PWeinstein PStates BHonkaniemi JSharp FR: Subarachnoid injections of lysed blood induce the hsp70 stress gene and produce DNA fragmentation in focal areas of the rat brain. Stroke 27:5045131996

    • Search Google Scholar
    • Export Citation
  • 21

    Matz PGFujimura MChan PH: Subarachnoid hemolysate produces DNA fragmentation in a pattern similar to apoptosis in mouse brain. Brain Res 858:3123192000

    • Search Google Scholar
    • Export Citation
  • 22

    Merry DEKorsmeyer SJ: Bcl-2 gene family in the nervous system. Annu Rev Neurosci 20:2452671997

  • 23

    Nau RHaase SBunkowski SBruck W: Neuronal apoptosis in the dentate gyrus in humans with subarachnoid hemorrhage and cerebral hypoxia. Brain Pathol 12:3293362002

    • Search Google Scholar
    • Export Citation
  • 24

    Park SYamaguchi MZhou CCalvert JWTang JZhang JH: Neurovascular protection reduces early brain injury after subarachnoid hemorrhage. Stroke 35:241224172004

    • Search Google Scholar
    • Export Citation
  • 25

    Prunell GFMathiesen TDiemer NHSvendgaard NA: Experimental subarachnoid hemorrhage: subarachnoid blood volume, mortality rate, neuronal death, cerebral blood flow, and perfusion pressure in three different rat models. Neurosurgery 52:1651662003

    • Search Google Scholar
    • Export Citation
  • 26

    Prunell GFSvendgaard NAAlkass KMathiesen T: Delayed cell death related to acute cerebral blood flow changes following subarachnoid hemorrhage in the rat brain. J Neurosurg 102:104610542005

    • Search Google Scholar
    • Export Citation
  • 27

    Richardson JT: Cognitive performance following rupture and repair of intracranial aneurysm. Acta Neurol Scand 83:1101221991

  • 28

    Schievink WIRiedinger MJhutty TKSimon P: Racial disparities in subarachnoid hemorrhage mortality: Los Angeles County, California, 1985–1998. Neuroepidemiology 23:2993052004

    • Search Google Scholar
    • Export Citation
  • 29

    Shih HCLin CLLee TYLee WSHsu C: 17beta-Estradiol inhibits subarachnoid hemorrhage-induced inducible nitric oxide synthase gene expression by interfering with the nuclear factor kappa B transactivation. Stroke 37:302530312006

    • Search Google Scholar
    • Export Citation
  • 30

    Springer JENottingham SAMcEwen MLAzbill RDJin Y: Caspase-3 apoptotic signaling following injury to the central nervous system. Clin Chem Lab Med 39:2993072001

    • Search Google Scholar
    • Export Citation
  • 31

    Toran-Allerand CD: Estrogen and the brain: beyond ER-alpha and ER-beta. Exp Gerontol 39:157915862004

  • 32

    Toung TJTraystman RJHurn PD: Estrogen-mediated neuroprotection after experimental stroke in male rats. Stroke 29:166616701998

  • 33

    Wise PM: Estrogens: protective or risk factors in brain function?. Prog Neurobiol 69:1811912003

  • 34

    Xing DFeng WMiller APWeathington NMChen YFNovak L: Estrogen modulates TNF-alpha-induced inflammatory responses in rat aortic smooth muscle cells through estrogen receptor-beta activation. Am J Physiol Heart Circ Physiol 292:H2607H26122007

    • Search Google Scholar
    • Export Citation
  • 35

    Zhou CYamaguchi MColohan ARZhang JH: Role of p53 and apoptosis in cerebral vasospasm after experimental subarachnoid hemorrhage. J Cereb Blood Flow Metab 25:5725822005

    • Search Google Scholar
    • Export Citation
  • 36

    Zhou CYamaguchi MKusaka GSchonholz CNanda AZhang JH: Caspase inhibitors prevent endothelial apoptosis and cerebral vasospasm in dog model of experimental subarachnoid hemorrhage. J Cereb Blood Flow Metab 24:4194312004

    • Search Google Scholar
    • Export Citation
  • 37

    Zubkov AYOgihara KBernanke DHParent ADZhang J: Apoptosis of endothelial cells in vessels affected by cerebral vasospasm. Surg Neurol 53:2602662000

    • Search Google Scholar
    • Export Citation
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