Effect of ketamine on apoptosis by energy deprivation in astroglioma cells using flow cytometry system.
Apoptosis is a programmed, physiologic mode of cell death that plays an important role in tissue homeostasis. As for the central nervous system, ischemic insults can induce pathophysiologic cascade of apoptosis in neurophils. Impairment of astrocyte functions during brain ischemia can critically influence neuron survival by neuronglia interactions. We aimed to elucidate the protective effect of ketamine on apoptosis by energy deprivation in astrocytes. Ischemic insults was induced with iodoacetate/ carbonylcyanide m-chlorophenylhydrazone (IAA/CCCP) 1.5 mM/20 microm or 150 microm/2 microm for 1 hr in the HTB-15 and CRL-1690 astrocytoma cells. Then these cells were reperfused with normal media or ketamine (0.1 mM) containing media for 1 hr or 24 hr. FITC-annexin-V staining and propidium iodide binding were determined by using flow cytometry. Cell size and granularity were measured by forward and side light scattering properties of flow cytometry system, respectively. An addition of ketamine during reperfusion increased the proportion of viable cells. Ketamine alleviated cell shrinkage and increased granularity during the early period, and ameliorated cell swelling during the late reperfusion period. Ketamine may have a valuable effect on amelioration of early and late apoptosis in the astrocytoma cells, even though the exact mechanism remains to be verified.
Choi
Soo Joo
SJ
Department of Anesthesiology and Pain Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
Kim
Myung Hee
MH
Lim
Seung Woon
SW
Gwak
Mi Sook
MS
D000778:Anesthetics, Dissociative / Q000494:pharmacology*; D017304:Annexin A5 / Q000494:pharmacology; D017209:Apoptosis; D001253:Astrocytes / Q000378:metabolism; D001254:Astrocytoma / Q000188:drug therapy* / Q000473:pathology; D001921:Brain / Q000473:pathology; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D045744:Cell Line, Tumor; D048429:Cell Size; D002470:Cell Survival; D002490:Central Nervous System / Q000187:drug effects / Q000473:pathology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007202:Indicators and Reagents / Q000494:pharmacology; D007461:Iodoacetates / Q000494:pharmacology; D007511:Ischemia / Q000473:pathology; D007649:Ketamine / Q000378:metabolism / Q000494:pharmacology*; D008027:Light; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D009504:Neutrophils / Q000378:metabolism; D010477:Perfusion; D011419:Propidium / Q000494:pharmacology; D012542:Scattering, Radiation; D013997:Time Factors; D014475:Uncoupling Agents / Q000494:pharmacology
D000778:Anesthetics, Dissociative; D017304:Annexin A5; D004791:Enzyme Inhibitors; D007202:Indicators and Reagents; D007461:Iodoacetates; D014475:Uncoupling Agents; D011419:Propidium; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D007649:Ketamine
Biotechniques. 1997 Sep;23(3):525-31
9298227
J Neurosci Res. 1997 Jun 15;48(6):563-70
9210526
Anesth Analg. 1998 Nov;87(5):1186-93
9806706
Histochem Cell Biol. 1998 Nov;110(5):467-76
9826126
Kidney Int. 1998 Dec;54(6):1955-66
9853260
Histochem Cell Biol. 1998 Dec;110(6):553-8
9860253
Brain Res Brain Res Protoc. 1999 Jul;4(2):209-16
10446416
Cytometry. 1998 Jan 1;31(1):1-9
9450519
Neuroreport. 1997 Dec 22;8(18):3871-5
9462458
Exp Neurol. 1998 Jan;149(1):51-63
9454614
Annu Rev Physiol. 2001;63:795-813
11181976
Glia. 2001 Aug;35(2):121-30
11460268
J Neurosci. 2001 Sep 1;21(17):6512-21
11517240
Neurochem Int. 2002 May;40(6):511-26
11850108
Brain Pathol. 2002 Oct;12(4):475-81
12408234
J Cereb Blood Flow Metab. 2003 Feb;23(2):137-49
12571445
J Neurosci. 2003 Apr 15;23(8):3364-72
12716944
Methods Mol Biol. 2003;228:271-9
12824560
Anal Biochem. 2004 Aug 15;331(2):385-94
15265746
Neuroscience. 1987 Jun;21(3):673-8
2819768
Brain Res. 1989 May 22;487(2):380-3
2567200
J Gen Physiol. 1990 May;95(5):837-66
2163431
Ann N Y Acad Sci. 1991;625:818-20
2058937
Neuroscience. 1991;45(2):461-5
1837070
Anesthesiology. 1992 May;76(5):755-62
1575344
J Cell Biol. 1992 Nov;119(3):493-501
1400587
FEBS Lett. 1994 Feb 14;339(1-2):40-4
8313978
J Immunol Methods. 1994 Jun 3;172(1):1-16
8207258
Stroke. 1994 Aug;25(8):1637-43
8042217
Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7162-6
7638161
J Exp Med. 1995 Nov 1;182(5):1545-56
7595224
Stroke. 1996 Apr;27(4):747-52
8614942
J Immunol. 1996 Jul 15;157(2):512-21
8752896
Cytometry. 1996 Jun 1;24(2):131-9
8725662
Anesthesiology. 1997 Feb;86(2):394-404
9054257
Anaesthesist. 1997 Mar;46 Suppl 1:S47-54
9163279
J Exp Med. 1997 Apr 21;185(8):1481-6
9126928
J Neurochem. 1998 Oct;71(4):1588-96
9751192