丹曲林可抑制经脂多糖和γ干扰素处理的大鼠肺泡巨噬细胞中的一氧化氮合酶。

Dantrolene inhibits nitric oxide synthase in rat alveolar macrophages treated with lipopolysaccharide and interferon-gamma.

作者信息

Li C Y, Chou T C, Wu C C, Wong C S, Ho S T, Yen M H, Ding Y A

机构信息

Department of Anesthesiology, Graduate Institute of Medical Sciences, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan, Republic of China.

出版信息

Can J Anaesth. 1998 Mar;45(3):246-52. doi: 10.1007/BF03012910.

DOI:10.1007/BF03012910

PMID:9579263

Abstract

PURPOSE

To examine the effects of dantrolene on nitric oxide (NO) production and on the activity and protein expression of inducible nitric oxide synthase (iNOS) induced by lipopolysaccharide (LPS) plus interferon-gamma (IFN gamma) in rat alveolar macrophages.

METHODS

Pulmonary alveolar macrophages isolated from Sprague-Dawley rats were used. After incubation of macrophages with dantrolene (1 to 100 microM) and LPS (1 microgram.ml-1) and IFN-gamma (100 u.ml-1) for 24 hr, the cell-free medium was removed for measuring the nitrite and tumour necrosis factor-alpha (TNF-alpha) levels by Griess reaction and ELISA kit, respectively. The harvested macrophages were also used to determine the activity of iNOS by using the conversion of [3H]-L-arginine to [3H]-L-citrulline method. Protein expression of iNOS was detected by Western blot analysis.

RESULTS

In rats alveolar macrophages, (i) dantrolene (1 to 100 microM) caused a dose-dependent suppression of the production of nitrite and TNF-alpha induced by LPS (1 microgram.ml-1) plus IFN-gamma (100 u.ml-1) and (ii) dantrolene (100 microM) inhibited the activity (by 37 +/- 5%, P < 0.01) and protein expression (by 39 +/- 12%, P < 0.01) of iNOS in response to LPS plus IFN-gamma.

CONCLUSION

Dantrolene inhibits NO production as well as the activity and expression of iNOS in alveolar macrophages treated with LPS plus IFN-gamma, which may be associated with the reduction of TNF-alpha production.

摘要

目的

研究丹曲林对脂多糖（LPS）加干扰素-γ（IFNγ）诱导的大鼠肺泡巨噬细胞中一氧化氮（NO）生成以及诱导型一氧化氮合酶（iNOS）活性和蛋白表达的影响。

方法

使用从Sprague-Dawley大鼠分离的肺泡巨噬细胞。将巨噬细胞与丹曲林（1至100微摩尔）、LPS（1微克/毫升）和IFNγ（100单位/毫升）孵育24小时后，去除无细胞培养基，分别通过格里斯反应和ELISA试剂盒测量亚硝酸盐和肿瘤坏死因子-α（TNF-α）水平。收获的巨噬细胞还用于通过[3H]-L-精氨酸转化为[3H]-L-瓜氨酸的方法测定iNOS的活性。通过蛋白质印迹分析检测iNOS的蛋白表达。

结果

在大鼠肺泡巨噬细胞中，（i）丹曲林（1至100微摩尔）对LPS（1微克/毫升）加IFNγ（100单位/毫升）诱导的亚硝酸盐和TNF-α生成产生剂量依赖性抑制，并且（ii）丹曲林（100微摩尔）抑制LPS加IFNγ刺激下iNOS的活性（降低37±5%，P<0.01）和蛋白表达（降低39±12%，P<0.01）。