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化学发光研究中溶剂二甲基亚砜使用情况的评估。

Evaluation of the use of the solvent dimethyl sulfoxide in chemiluminescent studies.

作者信息

Kahler C P

机构信息

Department of Pharmacology, Medical University of Southern Africa, Box 225, PO Medunsa 0204, South Africa.

出版信息

Blood Cells Mol Dis. 2000 Dec;26(6):626-33. doi: 10.1006/bcmd.2000.0340.

Abstract

Dimethyl sulfoxide (DMSO) is extensively used as solvent in in vitro chemiluminescent studies, to dissolve stimulants, drugs, and luminescent probes. DMSO is also known to scavenge hydroxyl radicals. In this study the effect of DMSO on the superoxide production of the different white blood cells, neutrophils, eosinophils, lymphocytes, and monocytes, after stimulation with phorbol myristate acetate (PMA), opsonized zymosan (OZ), and formylmethionylleucylphenylalanine (fMLP), was determined. Neutrophils, eosinophils, lymphocytes, and monocytes were isolated from human blood. DMSO was added in different concentrations before the cells were stimulated to produce superoxides. Concentrations of 10, 8, and 6% (v/v) DMSO cause significant inhibition of superoxide production (SP) after PMA and OZ stimulation. This was extended to 4 and 2% (v/v) DMSO after fMLP stimulation. The SP rates of eosinophils, lymphocytes, and monocytes are nonsignificantly decreased except at 10% (v/v) DMSO for lymphocytes after PMA stimulation and 10, 8, and 6% (v/v) DMSO for monocytes after fMLP stimulation. Due to strong scavenging effects of DMSO, this solvent should be used with caution as solvent in chemiluminescent studies. Concentrations of less than 1% (v/v) DMSO should not interfere with the results. During chemiluminescent studies care must be taken to ensure that the total amount of DMSO in the cuvette should not exceed 1% of the total volume, especially if the drug, the luminescent probe, and the stimulant were dissolved in DMSO.

摘要

二甲基亚砜(DMSO)在体外化学发光研究中被广泛用作溶剂,用于溶解刺激剂、药物和发光探针。已知DMSO还能清除羟基自由基。在本研究中,测定了DMSO对不同白细胞(中性粒细胞、嗜酸性粒细胞、淋巴细胞和单核细胞)在用佛波酯肉豆蔻酸酯乙酸酯(PMA)、调理酵母聚糖(OZ)和甲酰甲硫氨酰亮氨酰苯丙氨酸(fMLP)刺激后超氧化物生成的影响。从人血中分离出中性粒细胞、嗜酸性粒细胞、淋巴细胞和单核细胞。在细胞被刺激产生超氧化物之前,加入不同浓度的DMSO。10%、8%和6%(v/v)的DMSO浓度在PMA和OZ刺激后会显著抑制超氧化物生成(SP)。在fMLP刺激后,这一抑制作用扩展到4%和2%(v/v)的DMSO。嗜酸性粒细胞、淋巴细胞和单核细胞的SP率除了在PMA刺激后淋巴细胞为10%(v/v)DMSO以及fMLP刺激后单核细胞为10%、8%和6%(v/v)DMSO时无显著降低外,其他情况下均无显著降低。由于DMSO具有较强的清除作用,在化学发光研究中作为溶剂应谨慎使用。浓度低于1%(v/v)的DMSO不应干扰结果。在化学发光研究过程中,必须注意确保比色皿中DMSO的总量不超过总体积的1%,特别是当药物、发光探针和刺激剂溶解在DMSO中时。

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