国际麻醉学与复苏杂志   2014, Issue (9): 4-4
    
白细胞介素-10通过降低脂多糖诱导大鼠急性肺损伤高迁移率组蛋白1释放产生保护作用
张锋, 高鹏飞, 李紫薇, 武昊天, 王凯, 刘功俭1()
1.江苏省徐州医学院麻醉学重点实验室
The protective effect of interleukin-10 on lipopolysaccharide -induced acute lung injury in rats by attenuating the release of extracellular high mobility group protein 1
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摘要:

目的 旨在研究白细胞介素(interleukin, IL)-10对高迁移率组蛋白1(high mobility group protein 1, HMGB1)释放的影响及其肺保护作用,进而探讨IL-10在治疗急性肺损伤中的可能机制。 方法 采用腹腔注射脂多糖(lipopolysaccharide,LPS)致大鼠脓毒症急性肺损伤模型,健康SPF级雄性SD大鼠72只,体重180 g~220 g,采用随机数字表法,随机分为对照组即磷酸盐缓冲液(phosphate buffered solution, PBS)组(P组,6只)、急性肺损伤组即LPS组(L组,30只)、PBS+IL-10组(PI组,6只)、LPS+IL-10组(LI组,30只)。P组和PI组腹腔注射等体积PBS的同时分别经由气道滴注5 ml的PBS和IL-10,L组和LI组腹腔注射LPS的同时分别经由气道滴注等体积的PBS和IL-10。L组和LI组注射LPS后的4、8、16、24 h和48 h分别检测各组大鼠肺湿/干重比(wet/dry weight ratio, W/D)和支气管肺泡灌洗液(bronchoalveolar lavage fluid, BALF)中总蛋白浓度,酶联免疫吸附实验法检测BALF中炎性因子肿瘤坏死因子?蛳α(tumor necrosis factor-α, TNF-α)和IL?蛳6水平,苏木精-伊红(hematoxylin-eosin, HE)染色观察肺组织的病理变化,Western Blot分析肺组织中HMGB1表达水平。 结果 腹腔注射LPS后,L组大鼠肺组织W/D和BALF中总蛋白浓度分别为(5.68±0.12) mg/L和(254±105) mg/L,与P组比较,分别增加了12%和297%(P<0.05),BALF中TNF-α和IL-6水平明显增加(P<0.05),HE染色显示在注射LPS后4 h肺组织的细胞浸润达到峰值随后减少,肺组织中HMGB1水平升高(P<0.05);使用IL-10后,LI组肺组织W/D和BALF中总蛋白浓度分别为(5.28±0.14) mg/L和(109±48) mg/L,与L组比较,分别降低了7%和57%(P<0.05),BALF中炎性因子水平降低(P<0.05),肺组织细胞浸润改善,HMGB1表达下调(P<0.05)。 结论 IL-10对急性肺损伤具有保护作用,其机制可能为降低BALF中炎性因子的水平,下调晚期炎症介质HMGB1的表达,从而改善肺组织的细胞浸润实现的。

关键词: 白细胞介素-10; 脂多糖; 急性肺损伤; 高迁移率组蛋白1
Abstract:

Objective This study aimed to investigate the effect of interleukin(IL)-10 on extracellular release of high mobility group protein 1(HMGB1) in lipopolysaccharide(LPS)-induced acute lung injury in rats and its possible mechanism. Methods LPS(5 mg/kg,ip) induced acute lung injury model was used in our experiment, the healthy male Sprague-Dawley rats, weighed 180 g-220 g, were randomly divided into four groups: control group only used phosphate buffered solution (PBS) (group P, n=6), PBS+IL-10 group(group PI, n=6), acute lung injury group also LPS group(group L, n=30), LPS+interleukin-10 group( group LI, n=30). In the group L and LI, 4, 8, 16, 24 h and 48 h after the injection of LPS, the lung wet/dry weight ratio(W/D) and the concentration of the total protein in bronchoalveolar lavage fluid(BALF) were analyzed. The cytokines[tumor necrosis factor-α (TNF-α) and IL-6] in bronchoalveolar lavage fluid were detected by enzyme-linked immunosorbent assay. Hitophathological changes in lung tissue were observed by hematoxylin-eosin staining, and the expression of HMGB1 was analyzed by Western Blot. Results After using of LPS, the W/D and the concentration of the total protein in BALF in group L were(5.68±0.12) mg/L and(254±105)mg/L, which were increased by 12% and 297%(P<0.05),compared with the group P, the concentration of TNF-α and IL-6 in BALF, HE staining of the lung tissue infiltration and the expression of HMGB1 in the lung tissue were all increased(P<0.05). However after the intratracheal spray of IL-10,the W/D and the concentration of the total protein in BALF in group LI were (5.28±0.14) mg/L and(109±48) mg/L, which were decreased by 7% and 57%(P<0.05), compared with the group L, the level of inflammatory factors, lung tissue infiltration and the concentration of HMGB1 were all decreased (P<0.05). Conclusions IL-10 can attenuate the injury induced by LPS in acute lung injury. The mechanism may be due to the effect of decreasing the proinflammatory cytokines and down-regulating the expression of HMGB1.

Key words: Interleukin-10; Lipopolysaccharide; Acute lung injury; High mobility group protein 1