摘要
目的通过建立急性肺损伤大鼠模型和体外肺泡巨噬细胞培养体系,观察急性肺损伤中,肺泡巨噬细胞在氨溴索干预前后细胞因子表达的情况,探讨氨溴索可能的肺损伤保护机制。方法一、体内实验:大鼠24只,随机分成3组,每组8只:(1)正常对照组;(2)急性肺损伤组:采用腹腔注射酵母悬液方法复制大鼠急性肺损伤模型;(3)氨溴索治疗组:建立Au大鼠模型后用氨溴索干预。观察指标主要有:肺部病理学改变、测量大鼠肺系数、血氧分压,肺泡巨噬细胞TNF-α、IL-10、IL-24mRNA的表达则采用通过逆转录-聚合酶链式反应(RT-PCR)技术检测。二、体外实验:收集肺泡巨噬细胞后分3组:(1)正常对照组:肺泡巨噬细胞中加无菌生理盐水;(2)LPS组:予LPS 10mg/L刺激肺泡巨噬细胞;(3)LPS+氨溴索组:予LPS 10mg/L和氨溴索180μmol/L刺激肺泡巨噬细胞。分别于刺激前(0h),刺激后(6、12、24h)留取细胞。通过RT-PCR技术检测肺泡巨噬细胞TNF-α、IL-10、IL-24mRNA的表达变化。结果氨溴索治疗组在病理改变,肺系数,动脉血氧分压等各项指标均优于急性肺损伤组,氨溴索干预治疗组,TNF-α、IL-10、IL-24mRNA表达水平均明显下降,与急性肺损伤组比较差异有统计学意义(P〈0.05),但仍高于正常对照组。体外实验也得出了相同的结论。结论氨溴索可抑制急性肺损伤或LPS刺激的肺泡巨噬细胞TNF-α、IL-10、IL-24细胞因子mRNA的表达水平。
Objective To explore the mechanism of protection role of ambroxol against acute lung injury (ALI) by studying the change of cytokine mRNA expression during the course of ALI. Method The study was composed of experiments both in vivo and in vitro. ( 1 ) Experiments in vivo were as follows. Twenty-four sprague-Dawley rats were randomly divide into 3 groups, namely, control group (n = 8), acute lung injury group (LPS group, n = 8) and ambroxol group (LPS+ A group, n = 8). The rat model of acute lung injury was induced by intraperitoneal injection of 10 mmol/L lipopolysaccharide (LPS). The pathological alteration of lung tissue and arterial partial pressure of oxygen (PaO2) were observed. Expressions of TNF-α, IL-10 and IL- 24 mRNA were determined by using RT-PCR assay. (2) Experiments in vitro were the followings. Mveolar macrophage cells were collected and divided into 3 groups as above mentioned. Cells were treated with normal saline, with LPS, and with LPS plus ambroxol in dose of 180 μmol/L at 0, 6, 12 and 24 hours after exposure of LPS, respectively, in 3 groups as above stated. The expressions of TNF-α, IL-10 and IL-24 mRNA were also determine by using RT-PCR. Results The pathological changes of could be parially ameliorated by giving ambroxol. Massive hemorrhage along with vascular edema and infiltration occurred in the lungs of ALT rots. The pathological alterations in ALl rats treated with ambroxol were less severe. The expressions of TNF-α, IL-10 and IL-24 mRNA were dramatically increased in ALl rots, and were partially attenuated after treatment of ambroxol. Macrophages oxposed to LPS for 6 hours showed dramatical increase in expressions of TNF-α, IL-10 and IL-24 mRNA those remained at high levels afterwards. The expressions of TNF-α, IL-10 and IL-24 mRNA in macmphages after exposure to both LPS and ambroxol were increased less than those exposed to LPS alone. Conclusions Ambroxol can partially ameliorate the expressions of TNF-α IL-10 and IL-24 mRNA in alveolar macrophage induced by LPS.
出处
《中华急诊医学杂志》
CAS
CSCD
2008年第4期355-360,共6页
Chinese Journal of Emergency Medicine
基金
广西省自然科学基金项目(桂科自0447055)