CAJ | 학술논문

目的探讨肺血管内巨噬细胞(PIM)在感染性急性肺损伤(ALI)发病中的作用.方法采用改良Morton法分离、培养猪PIM;用贴壁法获得黏附的PIM,培养于RPMI 1640培养基,予10 mg/L脂多糖(LPS)刺激.用鼠胸腺细胞增殖法测定白细胞介素-1β(IL-1β)活性,酶联免疫吸附法(ELISA)测定IL-6、IL-8含量.结果 LPS刺激后,PIM释放IL-1β、IL-6和IL-8均增多,峰值分别出现在刺激后的2 h[IL-1β活性(10 400±2 389)次/min]、4 h[IL-6含量(0.80±0.36)μg/L]和6 h[IL-8含量(4.94±1.19)μg/L],与刺激前[IL-1β活性(213±85)次/min,IL-6含量(0.27±0.12)μg/L,IL-8含量(1.84±0.53)μg/L]比较差异均有统计学意义(均P＜0.01).结论 LPS刺激后,PIM分泌多种细胞因子,其中IL-1β升高最早,提示其在ALI发病早期起重要作用;而IL-6、IL-8升高较晚,且持续时间较长,可能对ALI的病情进展起重要作用.细胞因子间的相互作用在ALI的发病中似乎更为重要.
목적 탐토폐혈관내거서세포(PIM)재감염성급성폐손상(ALI)발병중적작용.방법 채용개량Morton법분리、배양저PIM;용첩벽법획득점부적PIM,배양우RPMI 1640배양기,여10 mg/L지다당(LPS)자격.용서흉선세포증식법측정백세포개소-1β(IL-1β)활성,매련면역흡부법(ELISA)측정IL-6、IL-8함량.결과 LPS자격후,PIM석방IL-1β、IL-6화IL-8균증다,봉치분별출현재자격후적2 h[IL-1β활성(10 400±2 389)차/min]、4 h[IL-6함량(0.80±0.36)μg/L]화6 h[IL-8함량(4.94±1.19)μg/L],여자격전[IL-1β활성(213±85)차/min,IL-6함량(0.27±0.12)μg/L,IL-8함량(1.84±0.53)μg/L]비교차이균유통계학의의(균P＜0.01).결론 LPS자격후,PIM분비다충세포인자,기중IL-1β승고최조,제시기재ALI발병조기기중요작용;이IL-6、IL-8승고교만,차지속시간교장,가능대ALI적병정진전기중요작용.세포인자간적상호작용재ALI적발병중사호경위중요.
Objective To investigate the role of pulmonary intravascular macrophages(PIMs)in the pathogenesis of acute lung injury(ALI)due to infection. Methods Porcine pulmonary blood vessels were flushed by modified Morton method, and PIMs were isolated and cultured. The adhered PIMs were collected with adhesion method and incubated in RPMI 1640 medium. They were challenged with lipopolysaccharide (LPS, 10 mg/L). The activity of interleukin-1β(IL-1β), and contents of IL-6 and IL-8 in the culture supernatant were measured by method of thymocyte proliferation and enzyme linked immunoadsorbent assay (ELISA). Results The released contents of IL-1β, IL-6 and IL-8 from PIMs were increased significantly compared with those before LPS challenge, and they peaked at 2 hours[IL-1β activity:(10 400±2 389)scintillant count/min], 4 hours[IL-6 content:(0. 80± 0. 36)μg/L], and 6 hours[IL-8 content:(4. 94± 1.19)μg/L]after LPS challenge, and the differences were significant compared with those before LPS challenge[IL-1β activity:(213±85)scintillant count/min, IL-6 content:(0. 27 ± 0. 12)μg/L, IL-8 content:(1.84±0.53)μg/L, all P＜0. 01]. Conclusion Among the cytokines released from PIMs after LPS challenge, the increase in IL-1β occurred earlier in comparison with that of IL-6 and IL-8, suggesting that the former might play an important role at the early stage of ALI; on the other hand, though the increase in IL-6 and IL-8 contents occurred later than that of IL-1β but it lasted for a longer duration,suggesting that they might be associated with the advancement of ALI. The results also suggested that interaction of these cytokines played a more important role in the pathogenesis of ALI.