氧化应激对牛阴茎退缩肌中一氧化氮能神经传递的抑制作用：超氧化物歧化酶模拟物的影响

Inhibition of nitrergic neurotransmission in the bovine retractor penis muscle by an oxidant stress: effects of superoxide dismutase mimetics.

作者信息

Mok J S, Paisley K, Martin W

机构信息

Clinical Research Initiative, Institute of Biomedical & Life Sciences, University of Glasgow.

出版信息

Br J Pharmacol. 1998 May;124(1):111-8. doi: 10.1038/sj.bjp.0701809.

DOI:10.1038/sj.bjp.0701809

PMID:9630350

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1565368/

Abstract

A number of superoxide dismutase (SOD) mimetics were examined both biochemically for their ability to inhibit the superoxide-catalyzed reduction of cytochrome c and nitro blue tetrazolium, and functionally for their ability to mimic authentic Cu/Zn SOD in restoring nitrergic neurotransmission in bovine retractor penis (BRP) muscle following its inhibition by oxidant stress. 2. The SOD mimetics investigated were CuSO4, MnCl2, CuDIPS (copper [II] [diisopropylsalicylate]2), MnTBAP (manganese [III] tetrakis 4-benzoic acid porphyrin), MnTMPyP (manganese [III] tetrakis 1-methyl-4-pyridyl porphyrin pentachloride), tiron (4,5-dihydroxy-1,3-benzene disulphonic acid), PTIYO (4-phenyl,2,2,5,5,-tetramethyl-3-imidazolin-1-yloxy-3-oxide) and tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl). 3. The rank order of potency in inhibiting the reduction of cytochrome c was: CuSO4 > or = MnCl2 > or = CuDIPS > or = MnTMPyP > MnTBAP > tempol > or = tiron > PTIYO. 4. The requirement for EDTA (0.1 mM) prevented assessment of the activity of CuSO4, MnCl2 and CuDIPS in the assay involving inhibition of reduction of nitro blue tetrazolium. However, the rank order of potency for those agents which could be examined (MnTMPyP > MnTBAP > tiron > or = tempol > PTIYO) was essentially similar to that seen in the cytochrome c assay. 5. Inhibition of endogenous Cu/Zn SOD with diethyldithiocarbamate (DETCA, 3 mM, 120 min) in BRP muscle strips, followed by addition of the superoxide anion generator, LY 83583 (1 microM), resulted in almost complete abolition of nitrergic relaxation (4 Hz, 10 s). 6. Authentic Cu/Zn SOD (1-300 u ml(-1)), CuSO4 (0.1-300 microM), MnCl2 (0.1-100 microM) and MnTMPyP (10-300 microM) each restored nitrergic transmission by around 50%. However, CuDIPS (0.1-30 microM), MnTBAP (0.1-100 microM), tempol (10 microM - 3 mM), PTIYO (1-300 microM) and tiron (10 microM - 10 mM) all failed to restore nitrergic transmission. 7. The ability of MnTMPyP to restore nitrergic neurotransmission may therefore provide a lead in the development of SOD mimetics as therapeutic agents in the treatment of neuropathies associated with oxidant stress.

摘要

对多种超氧化物歧化酶（SOD）模拟物进行了生物化学检测，以考察它们抑制超氧化物催化的细胞色素c和硝基蓝四唑还原的能力，并进行了功能检测，以考察它们在牛阴茎退缩肌（BRP）因氧化应激抑制后，模拟天然铜/锌超氧化物歧化酶恢复一氧化氮能神经传递的能力。2. 所研究的SOD模拟物有硫酸铜、氯化锰、CuDIPS（铜[II][二异丙基水杨酸酯]2）、MnTBAP（锰[III]四（4 - 苯甲酸）卟啉）、MnTMPyP（锰[III]四（1 - 甲基 - 4 - 吡啶基）卟啉五氯化物）、钛铁试剂（4,5 - 二羟基 - 1,3 - 苯二磺酸）、PTIYO（4 - 苯基，2,2,5,5 - 四甲基 - 3 - 咪唑啉 - 1 - 氧基 - 3 - 氧化物）和TEMPOL（4 - 羟基 - 2,2,6,6 - 四甲基哌啶 - N - 氧化物）。3. 抑制细胞色素c还原的效力排序为：硫酸铜≥氯化锰≥CuDIPS≥MnTMPyP＞MnTBAP＞TEMPOL≥钛铁试剂＞PTIYO。4. 在涉及抑制硝基蓝四唑还原的检测中，由于需要加入乙二胺四乙酸（EDTA，0.1 mM），因此无法评估硫酸铜、氯化锰和CuDIPS的活性。然而，对于那些能够检测的试剂，其效力排序（MnTMPyP＞MnTBAP＞钛铁试剂≥TEMPOL＞PTIYO）与细胞色素c检测中观察到的基本相似。5. 在BRP肌条中用二乙基二硫代氨基甲酸盐（DETCA，3 mM，120分钟）抑制内源性铜/锌超氧化物歧化酶，随后加入超氧阴离子生成剂LY 83583（1 microM），导致一氧化氮能舒张几乎完全消失（4 Hz，10秒）。6. 天然铜/锌超氧化物歧化酶（1 - 300单位/毫升）、硫酸铜（0.1 - 300 microM）、氯化锰（0.1 - 100 microM）和MnTMPyP（10 - 300 microM）各自使一氧化氮能传递恢复约50%。然而，CuDIPS（0.1 - 30 microM）、MnTBAP（0.1 - 100 microM）、TEMPOL（10 microM - 3 mM）、PTIYO（1 - 300 microM）和钛铁试剂（10 microM - 10 mM）均未能恢复一氧化氮能传递。7. 因此，MnTMPyP恢复一氧化氮能神经传递的能力可能为开发SOD模拟物作为治疗与氧化应激相关神经病变的治疗药物提供线索。