Nawrocka A, Papierz W, Bialasiewicz P, Stolarek R, Komos J, Nowak D
Department of Pathology, Medical University of Lódz, Czechoslowacka 8/10, Lódz, 92-216, Poland.
Pulm Pharmacol Ther. 1999;12(6):369-75. doi: 10.1006/pupt.1999.0219.
We have investigated whether pretreatment with N-acetylcysteine (NAC) and/or ambroxol (Amb), drugs known as reactive oxygen species (ROS) scavengers, would minimize lipopolysaccharide (LPS)-induced leucocyte accumulation in rat lung microvasculature and protect lungs from damage and the effect of these drugs on chemotactic peptide (fMLP)-induced chemiluminescence of human polymorphonuclear leukocytes (PMNs). Animals were injected ip with NAC (27.6 mg/kg, n=8), ambroxol (70 mg/kg, n=8), combination NAC+ambroxol (n=8), or 1 ml buffer alone (n=8), once a day for 3 consecutive days. Then animals were injected with LPS (17 mg/kg), and killed 3 h later. In each of another four groups eight rats were used as a control, and received the same drug treatment but LPS was replaced with 0.9% NaCl. PMNs and macrophages (Ms) were counted in histologic slides of lung tissue. Using computer image analysis we measured the area of alveolar profiles. Luminol-enhanced chemiluminescence was measured in PMNs suspensions obtained from healthy volunteers. Chemiluminescence intensity was measured in resting and fMLP-stimulated cells, and compared between cells incubated with Amb, NAC or distilled water. We observed significant differences in the number of PMNs and Ms, alveolar profile area between control and LPS-treated animals (P<0.01). PMNs and Ms were numerous in lungs of LPS-administered animals (PMNs: Median (M)=137.5 per 6 high power fields range (r)=54.0; Ms: M=123.0 r=11.0), less numerous in ambroxol-treated group (PMNs: M=101.5 r=32.0 and Ms:53.5 r=36.0), not abundant in NAC (PMNs:M=56.0 r=28.0 and Ms:M=20.5 r=13.0) and in NAC+ambroxol treated rats (PMNs:M=53.5 r=21.0 and Ms:M=29.0 r=9.0), and rare in LPS+drugs-untreated control group (PMNs:M=40.5 r=19.0 and Ms:M=18.5 r=15.0). Chemiluminescence assay revealed that 100 micro;M ambroxol stimulated fMLP-induced PMNs chemiluminescence and NAC of the same concentration had no significant effect.
In our experiment we showed that pretreatment with NAC and ambroxol may inhibit phagocyte influx to rat lung and may protect it from damage. We also revealed that NAC at dose 27.6 mg/kg has stronger protective properties than ambroxol at dose 70 mg/kg and this may result from enhancing effect of ambroxol on fMLP-provoked PMNs chemiluminescence.
我们研究了用N-乙酰半胱氨酸（NAC）和/或氨溴索（Amb）进行预处理，这两种药物作为活性氧（ROS）清除剂，是否能使脂多糖（LPS）诱导的白细胞在大鼠肺微血管中的积聚降至最低，并保护肺免受损伤，以及这些药物对趋化肽（fMLP）诱导的人多形核白细胞（PMN）化学发光的影响。动物腹腔注射NAC（27.6mg/kg，n = 8）、氨溴索（70mg/kg，n = 8）、NAC + 氨溴索组合（n = 8）或仅1ml缓冲液（n = 8），连续3天每天注射一次。然后给动物注射LPS（17mg/kg），3小时后处死。在另外四组中，每组8只大鼠作为对照，接受相同的药物治疗，但LPS被替换为0.9%氯化钠。在肺组织的组织学切片中对PMN和巨噬细胞（Ms）进行计数。使用计算机图像分析测量肺泡轮廓的面积。在从健康志愿者获得的PMN悬液中测量鲁米诺增强的化学发光。测量静息和fMLP刺激细胞中的化学发光强度，并在与Amb、NAC或蒸馏水孵育的细胞之间进行比较。我们观察到对照动物和LPS处理动物之间PMN和Ms的数量、肺泡轮廓面积存在显著差异（P < 0.01）。在给予LPS的动物肺中PMN和Ms数量众多（PMN：每6个高倍视野中位数（M）= 137.5，范围（r）= 54.0；Ms：M = 123.0，r = 11.0），在氨溴索处理组中数量较少（PMN：M = 101.5，r = 32.0；Ms：53.5，r = 36.0），在NAC处理组中不丰富（PMN：M = 56.0，r = 28.0；Ms：M = 20.5，r = 13.0），在NAC + 氨溴索处理的大鼠中也较少（PMN：M = 53.5，r = 21.0；Ms：M = 29.0，r = 9.0），在未用LPS和药物处理的对照组中很少见（PMN：M = 40.5，r = 19.0；Ms：M = 18.5，r = 15.0）。化学发光分析显示，100μM氨溴索刺激fMLP诱导的PMN化学发光，相同浓度的NAC没有显著影响。
在我们的实验中，我们表明用NAC和氨溴索预处理可能抑制吞噬细胞流入大鼠肺并可能保护其免受损伤。我们还发现，27.6mg/kg剂量的NAC比70mg/kg剂量的氨溴索具有更强的保护特性，这可能源于氨溴索对fMLP诱导的PMN化学发光的增强作用。
