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Effects of Reactive Oxygen Species and Nitrogen Species on the Excitability of Spinal Substantia Gelatinosa Neurons KCI 등재후보

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대한구강생물학회 (The Korean Academy of Oral Biology)
초록

Reactive oxygen species (ROS) and nitrogen species (RNS) are both important signaling molecules involved in pain transmission in the dorsal horn of the spinal cord. Xanthine oxidase (XO) is a well-known enzyme for the generation of superoxide anions (O2 ⦁-), while S-nitroso-N-acetyl-DLpenicillamine (SNAP) is a representative nitric oxide (NO) donor. In this study, we used patch clamp recording in spinal slices of rats to investigate the effects of O2 ⦁- and NO on the excitability of substantia gelatinosa (SG) neurons. We also used confocal scanning laser microscopy to measure XO- and SNAP-induced ROS and RNS production in live slices. We observed that the ROS level increased during the perfusion of xanthine and xanthine oxidase (X/XO) compound and SNAP after the loading of 2′,7′-dichlorofluorescin diacetate (H2DCF-DA), which is an indicator of intracellular ROS and RNS. Application of ROS donors such as X/XO, β -nicotinamide adenine dinucleotide phosphate (NADPH), and 3-morpholinosydnomimine (SIN-1) induced a membrane depolarization and inward currents. SNAP, an RNS donor, also induced membrane depolarization and inward currents. X/XO-induced inward currents were significantly decreased by pretreatment with phenyl N-tert-butylnitrone (PBN; nonspecific ROS and RNS scavenger) and manganese(III) tetrakis(4-benzoic acid) porphyrin (MnTBAP; superoxide dismutase mimetics). Nitro-L-arginine methyl ester (NAME; NO scavenger) also slightly decreased X/XO-induced inward currents, suggesting that X/XO-induced responses can be involved in the generation of peroxynitrite (ONOO-). Our data suggest that elevated ROS, especially O2 ⦁-, NO and ONOO-, in the spinal cord can increase the excitability of the SG neurons related to pain transmission.

목차
서 론
 재료 및 방법
  척수절편 제작
  실험용액
  전기생리학적 기록방법
  형광이미지 측정
  실험자료의 분석
 연구성적
  척수 후각세포에서 X/XO, SNAP 투여에 의한 활성산소량 변화
  척수 후각세포 흥분성에 대한 X/XO, SNAP 투여의효과
 고 찰
 References
저자
  • Sang Woo Chun(Dept. of Oral Physiology, College of Dentistry, Institute of Wonkwang Biomaterial and Implant, Wonkwang University, Iksan 570-749)
  • Areum Park(Dept. of Oral Physiology, College of Dentistry, Institute of Wonkwang Biomaterial and Implant, Wonkwang University, Iksan 570-749)
  • Joo Young Park(Dept. of Oral Physiology, College of Dentistry, Institute of Wonkwang Biomaterial and Implant, Wonkwang University, Iksan 570-749)