Neuroprotective effect of α-lipoic acid on cultured retinal ganglion cells under elevated pressure in vitro

( views:330, downloads:0 )
Author:
LIU Bing()
GUO Da-dong(Affiliated Eye Hospital of Shandong University of Traditional Chinese Medicine, Jinan 250014 , China)
BI Hong-sheng(Affiliated Eye Hospital of Shandong University of Traditional Chinese Medicine, Jinan 250014 , China)
()
()
()
Journal Title:
Chinese Journal of Ocular Fundus Diseases
Issue:
Volume 28, Issue 05, 2012
DOI:
10.3760/cma.j.issn.1005-1015.2012.05.017
Key Word:
Retinal ganglion cells/ pathology; Thioctic acid/therapeutic use; Cells, cultured;Animal experimentation

Abstract: Objective To observe the neuroprotective effect of α-lipoic acid (ALA) on cultured retinal ganglion cells (RGC-5) under elevated pressure in vitro.Methods Cultured RGC-5 cells were divided randomly into 4 groups,including normal control group (group A),negative control group (group B),elevated pressure group (group C) and elevated pressure + ALA group (group D).The cells of group A and C were not intervened with ALA.The cells of group B were treated with 200 μmol/L ALA.The cells of group D were treated with different concentrations of ALA (50,100,200 μmol/L) for one hour.Then cells of group C and D were exerted to 50 mm Hg (1 mm Hg=0.133 kPa) for 24 hours,while the cells of group A and B were exerted to normal pressures for 24 hours.The cell viability was measured using the methyl thiazolyl tetrazolium (MTT) assay and apoptosis was evaluated using 4',6 diamidino-2-phenylindole (DAPI) staining.Expression of MnSOD was determined by real-time polymerase chain reaction (RT-PCR)and Western blot,respectively.Results The cell viability of group B was (65.6 ± 3.4) %,which lower than that in group D of three concentrations of ALA[ (75.1± 3.3)%,(81.8 ± 2.9 ) %,(87.9 ± 3.1 ) % ],the differences were significantly (t =5.108,10.007,12.513; P<0.05).DAPI staining revealed that characteristic apoptotic changes,such as chromatin condensation,convoluted nuclei with cavitations,fragmentation of the nucleus,and apoptotic bodies appeared in RGC-5 cells after 24 ours pressure.There was almost no evidence of apoptosis in group D.RT-PCR and Western blot analysis revealed that the expression of MnSOD mRNA and protein were weakly expressed in group C compared with control A (t=22.045,26.979; P<0.01).Compared group C with group D,the level of MnSOD mRNA and protein in group D increased significantly (t=9.171,12.267,23.567,7.723,12.009,28.198;P<0.05).In addition,the presence of ALA was found to inhibit hydrostatic pressure induced damage of RGC-5 cells in a dose-dependent manner (F=134.273,194.597;P<0.01).Conclusion ALA can effectively improve the expression of MnSOD in RGC-5 cells under the condition of elevated pressure,enhance the ability of RGC-5 cells against oxidative damage.

  • [1]Izzotti A,Bagnis A,Saccd SC.The role of oxidative stress in glaucoma.Mutat Res,2006,612:105-114.
  • [2]Liu CL,Ogando D,Bonanno JA.SOD2 contributes to anti-oxidative capacity in rabbit corneal endothelial cells.Mol Vis,2011,17:2473-2481.
  • [3]Osborne NN.Pathogenesis of ganglion “cell death” in glaucoma and neuroprotection:focus on ganglion cell axonal mitochondria.Prog Brain Res,2008,173:339-352.
  • [4]Jia Z,Zhu H,Vitto MJ,et al.Alpha-lipoic acid potently inhibits peroxynitrite-mediated DNA strand breakage and hydroxyl radical formation:implications for the neuroprotective effects of alpha-lipoic acid.Mol Cell Biochem,2009,323:131-138.
  • [5]Liu Q,Ju WK,Crowston JG,et al.Oxidative stress is an early event in hydrostatic pressure-induced retinal ganglion cell damage.Invest Ophthalmol Vis Sci,2007,48:4580-4589.
  • [6]Ding JY,Kreipke CW,Speirs SL,et al.Hypoxia-inducible factor-1α signaling in aquaporin upregulation after traumatic brain injury.Neurosci Lett,2009,453:68-72.
  • [7]Ju WK,Liu Q,Kim KY,et al.Elevated hydrostatic pressure triggers mitochondrial fission and decreases cellular ATP in differentiated RGC-5 cells.Invest Ophthalmol Vis Sci,2007,48:2145-2151.
  • [8]Ju WK,Kim KY,Lindsey JD,et al.Elevated hydrostatic pressure triggers release of OPA1 and cytochrome C,and induces apoptotic cell death in differentiated RGC-5 cells.Mol Vis,2009,15:120-134.
  • [9]Orrenius S.Reactive oxygen species in mitochondria-mediated cell death.Drug Metab Rev,2007,39:443-455.
  • [10]Kowluru RA,Kowluru V,Xiong Y,et al.Overexpression of mitochondrial superoxide dismutase in mice protects the retina from diabetes-induced oxidative stress.Free Radic Biol Med,2006,41:1191-1196.
  • [11]Kanwar M,Chan PS,Kern TS,et al.Oxidative damage in the retinal mitochondria of diabetic mice:possible protection by superoxide dismutase.Invest Ophthalmol Vis Sci,2007,48:3805-3811.
  • [12]Lebovitz RM,Zhang H,Vogel H,et al.Neurodegeneration,myocardial injury,and perinatal death in mitochondrial superoxide dismutase-deficient mice.Proc Natl Acad Sci USA,1996,93:9782-9787.
  • [13]Toklu HZ,Hakan T,Celik H,et al.Neuroprotective effects of alpha-lipoic acid in experimental spinal cord injury in rats.J Spinal Cord Med,2010,33:401-409.
  • [14]Perera J,Tan JH,Jeevathayaparan S,et al.Neuroprotective effects of alpha lipoic acid on haloperidol-induced oxidative stress in the rat brain[J/OL].Cell Biosci,2011,1:E12[2011-03-22].http://www.cellandhioscience.com/content/1/1/12.
  • [15]Johnson TV,Tomarev SI.Rodent models of glaucoma.Brain Res Bull,2010,81:349-358.
  • [16]Lee JK,Lu S,Madhukar A.Real-time dynamics of Ca2+,caspase-3/7,and morphological changes in retinal ganglion cell apoptosis under elevated pressure[J/OL].PLoS ONE,2010,5:E13437[2010-10-18].http://www.plosone.org/article/info%3Adoi%2F10.1371% 2Fjournal.pone.0013437.
  • [17]Shimazawa M,Yamashima T,Agarwal N,et al.Neuroprotective effects of minocycline against in vitro and in vivo retinal ganglion cell damage.Brain Res,2005,1053:185-194.
  • [18]Kumar DM,Perez E,Cai ZY,et al.Role of nonfeminizing estrogen analogues in neuroprotection of rat retinal ganglion cells against glutamate-induced cytotoxicity.Free Radic Biol Med,2005,38:1152-1163.
  • [19]Agar A,Li S,Agarwal N,et al.Retinal ganglion cell line apoptosis induced by hydrostatic pressure.Brain Res,2006,1086:191-200.
WanfangData CO.,Ltd All Rights Reserved
About WanfangData | Contact US
Healthcare Department, Fuxing Road NO.15, Haidian District Beijing, 100038 P.R.China
Tel:+86-010-58882616 Fax:+86-010-58882615 Email:yiyao@wanfangdata.com.cn