丹曲林对内毒素诱导的白细胞介素-10、肿瘤坏死因子-α及一氧化氮在体内和体外产生的调节作用

Modulation by dantrolene of endotoxin-induced interleukin-10, tumour necrosis factor-alpha and nitric oxide production in vivo and in vitro.

作者信息

Haskó G, Szabó C, Németh Z H, Lendvai B, Vizi E S

机构信息

Department of Pharmacology, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest.

出版信息

Br J Pharmacol. 1998 Jul;124(6):1099-106. doi: 10.1038/sj.bjp.0701934.

DOI:10.1038/sj.bjp.0701934

PMID:9720779

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1565490/

Abstract

Intracellular calcium has been suggested to be an important mediator of the cellular response in endotoxaemia and shock. Dantrolene is an agent that interferes with intracellular calcium fluxes resulting in a decreased availability of calcium in the cytoplasm. Here we have investigated the effect of dantrolene on lipopolysaccharide (LPS)-induced production of interleukin-10 (IL-10), tumour necrosis factor-alpha (TNF-alpha), and nitric oxide (NO) in mice and in cultured RAW 264.7 macrophages in vitro. 2. In BALB/c mice, LPS-induced plasma IL-10 levels were significantly enhanced by pretreatment with dantrolene (20 mg kg(-1), i.p.) (P < 0.005 at the 90 min time-point). On the other hand, dantrolene pretreatment suppressed circulating TNF-alpha and nitrite/nitrate (breakdown products of NO) concentrations. However, dantrolene had no effect on LPS-induced plasma interleukin-6 (IL-6) levels (67.22+/-5.51 ng ml(-1) in vehicle-pretreated mice and 62.22+/-3.66 ng ml(-1) in dantrolene-pretreated mice, n = 9). 3. Dantrolene inhibited TNF-alpha and NO production in C57BL/6 IL-10+/+ mice, as well as in their IL-10 deficient counterparts (C57BL/6 IL-10(0/0)). 4. In RAW 264.7 macrophages, dantrolene (10-300 microM) reduced IL-10, TNF-alpha, and nitrite (breakdown product of NO) production elicited by LPS (10 microg ml(-1)). Dantrolene (300 microM) did not affect the LPS-induced nuclear translocation of transcription factor nuclear factor kappaB in these cells. 5. Although LPS failed to alter the intracellullar concentration of calcium in single macrophages loaded with Fura-2, dantrolene caused a significant decrease of the basal calcium level as determined 30 min after dantrolene treatment (P < 0.005). ATP (1 mM) caused a rapid rise in intracellular calcium levels in both dantrolene-pretreated and vehicle-pretreated cells. 6. These results indicate that unlike the secretion of TNF-alpha and NO, IL-10 production is differentially regulated in vitro and in vivo. The decrease of plasma levels of the pro-inflammatory mediators TNF-alpha and NO, and increase in circulating IL-10 concentrations by dantrolene suggest that this drug might offer a new therapeutic approach in inflammatory diseases and septic shock.

摘要

细胞内钙已被认为是内毒素血症和休克中细胞反应的重要介质。丹曲林是一种干扰细胞内钙通量的药物，导致细胞质中钙的可用性降低。在此，我们研究了丹曲林对小鼠和体外培养的RAW 264.7巨噬细胞中脂多糖（LPS）诱导的白细胞介素-10（IL-10）、肿瘤坏死因子-α（TNF-α）和一氧化氮（NO）产生的影响。2. 在BALB/c小鼠中，丹曲林（20 mg kg⁻¹，腹腔注射）预处理可显著提高LPS诱导的血浆IL-10水平（在90分钟时间点P < 0.005）。另一方面，丹曲林预处理可抑制循环中的TNF-α和亚硝酸盐/硝酸盐（NO的分解产物）浓度。然而，丹曲林对LPS诱导的血浆白细胞介素-6（IL-6）水平没有影响（载体预处理小鼠中为67.22±5.51 ng ml⁻¹，丹曲林预处理小鼠中为62.22±3.66 ng ml⁻¹，n = 9）。3. 丹曲林抑制C57BL/6 IL-10⁺/⁺小鼠以及它们的IL-10缺陷对应物（C57BL/6 IL-10(0/0)）中TNF-α和NO的产生。4. 在RAW 264.7巨噬细胞中，丹曲林（10 - 300 μM）可降低LPS（10 μg ml⁻¹）诱导的IL-10、TNF-α和亚硝酸盐（NO的分解产物）的产生。丹曲林（300 μM）对这些细胞中LPS诱导的转录因子核因子κB的核转位没有影响。5. 尽管LPS未能改变加载Fura-2的单个巨噬细胞内的钙浓度，但丹曲林处理30分钟后测定显示，丹曲林可导致基础钙水平显著降低（P < 0.005）。ATP（1 mM）可使丹曲林预处理和载体预处理的细胞内钙水平迅速升高。6. 这些结果表明，与TNF-α和NO的分泌不同，IL-10的产生在体外和体内受到不同的调节。丹曲林降低促炎介质TNF-α和NO的血浆水平，并增加循环中IL-10的浓度，这表明该药物可能为炎症性疾病和脓毒性休克提供一种新的治疗方法。