BMC Neuroscience | |
The effects of different doses of estradiol (E2) on cerebral ischemia in an in vitro model of oxygen and glucose deprivation and reperfusion and in a rat model of middle carotid artery occlusion | |
Li-Ze Xiong2  Wu-Gang Hou2  Hai-Long Dong2  Shi-Quan Wang2  Lan Shen1  Yan Li1  Pei Qin2  Yu-Long Ma2  | |
[1] Department of Biochemistry and Molecular Biology, The State Key Laboratory of Cancer Biology, The Fourth Military Medical University, Xi’an 710032, P R China;Department of Anesthesiology, Xijing Hospital, The Fourth Military Medical University, Xi’an 710032, P R China | |
关键词: Oxygen and glucose deprivation (OGD); Middle carotid artery occlusion (MCAO); Ischemia; Neuroprotection; Estrogen; | |
Others : 1140001 DOI : 10.1186/1471-2202-14-118 |
|
received in 2013-03-25, accepted in 2013-09-25, 发布年份 2013 | |
【 摘 要 】
Background
Because neuroprotective effects of estrogen remain controversial, we aimed to investigate the effect of different doses of estradiol (E2) on cerebral ischemia using both in vivo and in vitro experiments.
Results
PC12 cells were cultured at physiological (10 nM and 20 nM) or pharmacological (10 μM and 20 μM) dosages of E2 for 24 hours (h). The results of 5-bromodeoxyuridine (Brdu) incorporation and flow cytometric analysis showed that physiological doses of E2 enhanced cell proliferation and pharmacological doses of E2 inhibited cell proliferation. After the cells were exposed to oxygen and glucose deprivation (OGD) for 4 h and reperfusion for 20 h, the results of 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyl tetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release assay, flow cytometric analysis and Western blot analysis showed that physiological doses of E2 enhanced cell viability, reduced cell apoptosis and decreased the expression of pro-apoptotic protein caspase-3. In contrast, pharmacological doses of E2 decreased cell viability and induced cell apoptosis. In vivo, adult ovariectomized (OVX) female rats received continuous subcutaneous injection of different doses of E2 for 4 weeks. Transient cerebral ischemia was induced for 2 h using the middle cerebral artery occlusion (MCAO) technique, followed by 22 h of reperfusion. The results of Garcia test, 2, 3, 5-triphenyltetrazolium chloride (TTC) staining showed that 6 μg/kg and 20 μg/kg E2 replacement induced an increase in neurological deficit scores, a decrease in the infarct volume and a reduction in the expression of caspase-3 when compared to animals in the OVX group without E2 treatment. However, 50 μg/kg E2 replacement treatment decreased neurological deficit scores, increased the infarct volume and the expression of caspase-3 when compared to animals in the control group and 6 up/kg or 20 μg/kg E2 replacement group.
Conclusion
We conclude that physiological levels of E2 exhibit neuroprotective effects on cerebral ischemia; whereas, pharmacological or supraphysiological doses of E2 have damaging effects on neurons after cerebral ischemia.
【 授权许可】
2013 Ma et al.; licensee BioMed Central Ltd.
【 预 览 】
Files | Size | Format | View |
---|---|---|---|
20150324043146142.pdf | 2477KB | download | |
Figure 9. | 48KB | Image | download |
Figure 8. | 111KB | Image | download |
Figure 7. | 126KB | Image | download |
Figure 6. | 44KB | Image | download |
Figure 5. | 139KB | Image | download |
Figure 4. | 45KB | Image | download |
Figure 3. | 106KB | Image | download |
Figure 2. | 182KB | Image | download |
Figure 1. | 171KB | Image | download |
【 图 表 】
Figure 1.
Figure 2.
Figure 3.
Figure 4.
Figure 5.
Figure 6.
Figure 7.
Figure 8.
Figure 9.
【 参考文献 】
- [1]Dubal DB, Wise PM: Neuroprotective effects of estradiol in middle-aged female rats. Endocrinology 2001, 142(1):43-48.
- [2]Saleh TM, Cribb AE, Connell BJ: Estrogen-induced recovery of autonomic function after middle cerebral artery occlusion in male rats. Am J Physiol Regul Integr Comp Physiol 2001, 281(5):R1531-1539.
- [3]Viscoli CM, Brass LM, Kernan WN, Sarrel PM, Suissa S, Horwitz RI: A clinical trial of estrogen-replacement therapy after ischemic stroke. N Engl J Med 2001, 345(17):1243-1249.
- [4]Rossouw JE, Anderson GL, Prentice RL, LaCroix AZ, Kooperberg C, Stefanick ML, Jackson RD, Beresford SA, Howard BV, Johnson KC, Kotchen JM, Ockene J, Writing Group for the Women’s Health Initiative Investigators: Risks and benefits of estrogen plus progestin in healthy postmenopausal women: principal results from the women’s health initiative randomized controlled trial. JAMA 2002, 288(3):321-333.
- [5]Strom JO, Theodorsson E, Holm L, Theodorsson A: Different methods for administering 17b-estradiol to ovariectomized rats result in opposite effects on ischemic brain damage. BMC Neurosci 2010, 11:39. BioMed Central Full Text
- [6]Strom JO, Theodorsson A, Theodorsson E: Dose-related neuroprotective versus neurodamaging effects of estrogens in rat cerebral ischemia: a systematic analysis. J Cereb Blood Flow Metab 2009, 29(8):1359-1372.
- [7]Prewitt AK, Wilson ME: Changes in estrogen receptor-alpha mRNA in the mouse cortex during development. Brain Res 2007, 1134(1):62-69.
- [8]Solum DT, Handa RJ: Localization of estrogen receptor alpha (ER [alpha]) in pyramidal neurons of the developing rat hippocampus. Brain Res Dev Brain Res 2001, 128(2):165-175.
- [9]Toung TJ, Traystman RJ, Hurn PD: Estrogen-mediated neuroprotection after experimental stroke in male rats. Stroke 1998, 29(8):1666-1670.
- [10]Yang SH, Liu R, Wu SS, Simpkins JW: The use of estrogens and related compounds in the treatment of damage from cerebral ischemia. Ann N Y Acad Sci 2003, 1007:101-107.
- [11]Adams KL, Maxson MM, Mellander L, Westerink RH, Ewing AG: Estradiol inhibits depolarization-evoked exocytosis in PC12 cells via N-type voltage-gated calcium channels. Cell Mol Neurobiol 2010, 30(8):1235-1242.
- [12]Majewski Ł, Sobczak M, Wasik A, Skowronek K, Rędowicz MJ: Myosin VI in PC12 cells plays important roles in cell migration and proliferation but not in catecholamine secretion. J Muscle Res Cell Motil 2011, 32(4–5):291-302.
- [13]Shen H, Yuan Y, Ding F, Liu J, Gu X: The protective effects of achyranthes bidentata polypeptides against NMDA-induced cell apoptosis in cultured hippocampal neurons through differential modulation of NR2A- and NR2B-containing NMDA receptors. Brain Res Bull 2008, 77(5):274-281.
- [14]Deng X, Luan Q, Chen W, Wang Y, Wu M, Zhang H, Jiao Z: Nanosized zinc oxide particles induce neural stem cell apoptosis. Nanotechnology 2009, 20(11):115101.
- [15]Lasota A, Danowska-Klonowska D: Experimental osteoporosis-different methods of OVX in female white rats. Rocz Akad Med Bialymst 2004, 49(Suppl 1):129-131.
- [16]Inderdeo DS, Edwards DR, Han VK, Khokha R: Temporal and spatial expression of tissue inhibitors of metalloproteinases during the natural ovulatory cycle of the mouse. Biol Reprod 1996, 55(3):498-508.
- [17]Dundar SO, Ozcura F, Cetin ED, Beder N, Dundar M: Effects of estrogen replacement therapy on vascular endothelial growth factor expression in choroidal and retinal vasculature. Bratisl Lek Listy 2010, 111(9):473-476.
- [18]Longa EZ, Weinstein PR, Carlson S, Cummins R: Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 1989, 20(1):84-91.
- [19]Wang Y, Hayashi T, Chang CF, Chiang YH, Tsao LI, Su TP, Borlongan C, Lin SZ: Methamphetamine potentiates ischemia/reperfusion insults after transient middle cerebral artery ligation in mice. Stroke 2001, 32(3):775-782.
- [20]Garcia JH, Wagner S, Liu KF, Hu XJ: Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rats. Stroke 1995, 26(4):627-634.
- [21]Wang Y, Chang CF, Morales M, Chiang YH, Harvey BK, Su TP, Tsao LI, Chen S, Thiemermann C: Diadenosine tetraphosphate protects against injuries induced by ischemia and 6-hydroxydopamine in rat brain. J Neurosci 2003, 23(21):7958-7965.
- [22]McEwen BS, Alves SE: Estrogen actions in the central nervous system. Endocr Rev 1999, 20(3):279-307.
- [23]Wang JM, Liu L, Brinton RD: Estradiol-17-induced human neural progenitor cell proliferation is mediated by an estrogen receptor-phosphorylated extracellularly regulated kinase pathway. Endocrinology 2008, 149(1):208-218.
- [24]Mérot Y, Ferrière F, Gailhouste L, Huet G, Percevault F, Saligaut C, Flouriot G: Different outcomes of unliganded and liganded estrogen receptor-αon neurite utgrowth in PC12 Cells. Endocrinology 2009, 150(1):200-211.
- [25]Wang JM, Johnston PB, Ball BG, Brinton RD: The neurosteroid allo-pregnanolone promotes proliferation of rodent and human neural progenitor cells and regulates cell-cycle gene and protein expression. J Neurosci 2005, 25(19):4706-4718.
- [26]Masui Y: From oocyte maturation to the in vitro cell cycle: the history of discoveries of maturation-promoting factor (MPF) and cytostatic factor (CSF). Differentiation 2001, 69(1):1-17.
- [27]Jung JY, Roh KH, Jeong YJ, Kim SH, Lee EJ, Kim MS, Oh WM, Oh HK, Kim WJ: Estradiol protects PC12 cells against CoCl -induced apoptosis. Brain Res Bull 2008, 76(6):579-585.
- [28]Jover T, Tanaka H, Calderone A, Oguro K, Bennett MV, Etgen AM, Zukin RS: Estrogen protects against global ischemia-induced neuronal death and prevents activation of apoptotic signaling cascades in the hippocampal CA1. J Neurosci 2002, 22(6):2115-2124.
- [29]Barha CK, Dalton GL, Galea LA: Low doses of 17a-estradiol and 17b-estradiol facilitate, whereas higher doses of estrone and 17a- and 17b-estradiol impair, contextual fear conditioning in adult female rats. Neuropsychopharmacology 2010, 35(2):547-559.
- [30]Heldring N, Pike A, Andersson S, Matthews J, Cheng G, Hartman J, Tujague M, Ström A, Treuter E, Warner M, Gustafsson JA: Estrogen receptors: how do they signal and what are their targets. Physiol Rev 2007, 87(3):905-931.
- [31]Hall JM, Couse JF, Korach KS: The multifaceted mechanisms of estradiol and estrogen receptor signaling. J Biol Chem 2001, 276(40):36869-36872.
- [32]Chu Z, Andrade J, Shupnik MA, Moenter SM: Differential regulation of gonadotropin-releasing hormone neuron activity and membrane properties by acutely applied estradiol: dependence on dose and estrogen receptor subtype. J Neurosci 2009, 29(17):5616-5627.
- [33]Funakoshi T, Yanai A, Shinoda K, Kawano MM, Mizukami Y: G protein-coupled receptor 30 is an estrogen receptor in the plasma membrane. Biochem Biophys Res Commun 2006, 346(3):904-910.
- [34]Gingerich S, Kim GL, Chalmers JA, Koletar MM, Wang X, Wang Y, Belsham DD: Estrogen receptor alpha and G-protein coupled receptor 30 mediate the neuroprotective effects of 17beta-estradiol in novel murine hippocampal cell models. Neuroscience 2010, 170(1):54-66.
- [35]Lebesgue D, Traub M, De Butte-Smith M, Chen C, Zukin RS, Kelly MJ, Etgen AM: Acute administration of non-classical estrogen receptor agonists attenuates ischemia-induced hippocampal neuron loss in middle-aged female rats. PLoS One 2010, 5(1):e8642.
- [36]Liu SB, Zhang N, Guo YY, Zhao R, Shi TY, Feng SF, Wang SQ, Yang Q, Li XQ, Wu YM, Ma L, Hou Y, Xiong LZ, Zhang W, Zhao MG: G-protein coupled receptor 30 mediates rapid neuroprotective effects of estrogen via depression of NR2B-containing NMDA receptors. J Neurosci 2012, 32(14):4887-4900.
- [37]Jover-Mengual T, Miyawaki T, Latuszek A, Alborch E, Zukin RS, Etgen AM: Acute estradiol protects CA1 neurons from ischemia-induced apoptotic cell death via the PI3K/Akt pathway. Brain Res 2010, 1321:1-12.
- [38]Hojo Y, Higo S, Ishii H, Ooishi Y, Mukai H, Murakami G, Kominami T, Kimoto T, Honma S, Poirier D, Kawato S: Comparison between hippocampus-synthesized and circulation-derived sex steroids in the hippocampus. Endocrinology 2009, 150(11):5106-5112.
- [39]Mukai H, Tsurugizawa T, Ogiue-Ikeda M, Murakami G, Hojo Y, Ishii H, Kimoto T, Kawato S: Local neurosteroid production in the hippocampus: influence on synaptic plasticity of memory. Neuroendocrinology 2006, 84(4):255-263.