Wang Zong-Yu, Yang Ba-Xian, Zhu Xi, Wu Sheng-Nan
Department of Anesthesiology, Peking University People's Hospital, Beijing 100044, China.
Zhongguo Wei Zhong Bing Ji Jiu Yi Xue. 2011 Apr;23(4):239-42.
To observe the local changes in alveoli in intravenous endotoxin induced acute lung injury (ALI) rat model after inhalation of aerosolized unfractioned heparin (UFH), and to observe its effects on coagulability, fibrinolysis and inflammatory response.
Eighty seven male Wistar rats were divided into groups according to table of random number: sham, model, heparin therapy (HT) and heparin prophylaxis (HP). Endotoxin induced ALI model was reproduced by intravenous administration of lipopolysaccharide (LPS). Rats in HP and HT groups received aerosolized UFH before and after injection with LPS respectively, while rats in both sham and model groups inhaled aerosolized normal saline. Rats in each group were respectively sacrificed at 6, 12 and 24 hours after intravenous administration of LPS, and bronchoalveolar lavage fluid (BALF) was collected. Enzyme linked immunosorbent assay was used to measure the level of thrombin antithrombin (TAT), tissue type plasminogen activator (t-PA), urokinase type plasminogen activator (u-PA), plasminogen activator inhibitor 1 (PAI-1), tumor necrosis factor α (TNF-α), interleukin 6 (IL-6) in BALF.
At 6 hours after injury, the level of TAT (μg/L) in model group (3.346±0.585) was highest, that in HT group (2.764±0.100) was higher, and that in HP group (2.564±0.216) was lowest in BALF (all P<0.05). The t-PA (μg/L) concentration in HP group (3.037±0.524) was highest, that in HT group (2.494±0.191) was higher, and that in model group (1.716±0.125) was lowest (all P<0.05). Compared with model group, u-PA (μg/L) level in HP group dramatically enhanced (0.411±0.118 vs. 0.303±0.049, P<0.05). The concentration of PAI-1 (μg/L) in HP group was significantly lower than that of HT and model groups (2.296±0.246 vs. 2.597±0.425, 2.834±0.198, both P<0.05). In HP group, it was still lower than that in HT group at 12 hours (1.273±0.441 vs. 1.817±0.252, P<0.05). TNF-α (ng/L) levels in HT and HP groups were markedly lower compared with model group (68.154±3.915, 36.990±6.539 vs. 77.001±4.485) at 6 hours, and the level in HP group was lower than that in HT group (all P<0.05). TNF-α concentration in HP group was still the lowest at 12 hours (15.287±4.754), and there was significant difference compared with HT and model groups (26.756±5.336, 23.674±4.398, both P<0.05). The levels of IL-6 were not distinctively different among model, HT and HP groups at various time points.
It was proved that inhalation of aerosolized UFH resulted in lowering local coagulability, alleviating fibrinolytic depression, improving fibrinolysis , and attenuating inflammation in endotoxin induced ALI rat model. More prominent results will be obtained when it was use as a prophylactic measure. The optimal time of usage is 6 hours after endotoxin injection.
观察雾化吸入未分级肝素（UFH）后静脉注射内毒素致急性肺损伤（ALI）大鼠模型肺泡局部的变化，观察其对凝血、纤溶及炎症反应的影响。
87只雄性Wistar大鼠按随机数字表分为假手术组、模型组、肝素治疗组（HT）和肝素预防组（HP）。通过静脉注射脂多糖（LPS）复制内毒素诱导的ALI模型。HP组和HT组大鼠分别在注射LPS前后雾化吸入UFH，假手术组和模型组大鼠吸入雾化生理盐水。静脉注射LPS后6、12和24小时分别处死每组大鼠，收集支气管肺泡灌洗液（BALF）。采用酶联免疫吸附测定法检测BALF中凝血酶抗凝血酶（TAT）、组织型纤溶酶原激活物（t-PA）、尿激酶型纤溶酶原激活物（u-PA）、纤溶酶原激活物抑制剂1（PAI-1）、肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）水平。
损伤后6小时，模型组BALF中TAT（μg/L）水平最高（3.346±0.585），HT组（2.764±0.100）较高，HP组（2.564±0.216）最低（均P<0.05）。HP组t-PA（μg/L）浓度最高（3.037±0.524），HT组（2.494±0.191）较高，模型组（1.716±0.125）最低（均P<0.05）。与模型组比较，HP组u-PA（μg/L）水平显著升高（0.411±0.118比0.303±0.049，P<0.05）。HP组PAI-1（μg/L）浓度显著低于HT组和模型组（2.296±0.246比2.597±0.425、2.834±0.198，均P<0.05）。HP组在12小时时仍低于HT组（1.273±0.441比1.817±0.252，P<0.05）。6小时时，HT组和HP组TNF-α（ng/L）水平显著低于模型组（68.154±3.915、36.990±6.539比77.001±4.485），且HP组低于HT组（均P<0.05）。HP组TNF-α浓度在12小时时仍最低（15.287±4.754），与HT组和模型组比较差异有统计学意义（26.756±5.336、23.674±4.398，均P<0.05）。各时间点模型组、HT组和HP组IL-6水平差异无统计学意义。
证明雾化吸入UFH可降低内毒素诱导的ALI大鼠模型的局部凝血功能，减轻纤溶抑制，改善纤溶，并减轻炎症。作为预防措施使用时效果更显著。最佳使用时间为内毒素注射后6小时。
