贝母素甲对急性肺损伤小鼠TNF-α、IL-4、IL-10及PGE_2时空表达的影响

An effect of PM on expression of TNF-α, IL-4, IL-10 and PGE_2 in mice with acute lung injury

目的:本研究探讨贝母素甲(PM)对脂多糖(LPS)诱导的小鼠急性肺损伤过程中TNF-α、IL-4、IL-10及PGE_2时间和空间表达的影响。方法:选取60只SPF级小鼠,体重25~30g,随机分成正常组、模型组、阳性对照组(Dex组)、贝母素甲(PM)低、中、高剂量组,腹腔注射戊巴比妥(40mg/kg)麻醉小鼠,模型组、Dex组及PM组小鼠给予气管内注射4mg/kg LPS诱导建立急性肺损伤模型。以LPS注入前1h、12h、24h、36h和注入后12h、24h、36h作为时间点,分别给予五组动物生理盐水、10mg/kg醋酸地塞米松、0.1mg/kg PM溶液、1mg/kg PM溶液、10mg/kg PM溶液,正常组给予同等体积生理盐水。实验结束后解剖肺脏,观察肺脏形态学改变,分别用Real-Time PCR和酶免疫吸附实验(ELISA)测定肿瘤坏死因子(TNF-α)、IL-4、IL-10的m RNA水平和蛋白含量及血液PGE_2的水平。结果:与正常组和阳性对照组比较,LPS注射导致肺部出现明显的水肿和炎症细胞浸润等现象,TNF-α、PGE_2含量分别增加了145.1%(P<0.05)、64.5%(P<0.05);与模型组比较,PM处理后LPS对肺泡细胞的破坏明显减轻,TNF-α、PGE_2含量分别降低了80.4%(P<0.05)、35.6%(P<0.05),但IL-4和IL-10蛋白水平分别增加了2.9倍(P<0.01)和2.1倍(P<0.01)。结论:PM处理后使肺组织中TNF-α、PGE_2含量明显降低,LPS对肺泡细胞的破坏显著减轻,对急性肺损伤小鼠肺组织具有保护作用。

Objective: To investigate efficacy of PM on expression of TNF-α, IL-4, IL-10 and PGE2 in mice with acute lung injury from LPS. Methods: 60 mice with SPF were randomly divided into the normal group, model group, positive control group (Dex group) and PM low, middle and high dose groups. All mice were intraperitoneally injected pentobarbital (40mg/kg). The model group, Dex group and PM groups were intratracheally injected LPS (4mg/kg) to establish acute lung injury model. Before and after injecting LPS 1 hour, 12 hours, 24 hours, 36 hours, the model group, Dex group and PM groups were given saline, 10mg/kg dexamethasone hydrochloride, 0.1mg/kg PM, 1mg/kg PM, 10mg/kg PM; the normal group was given the same volume of saline. At the end of the experiment, the lungs were dissected, and morphological changes were observed. TNF-α, IL-4 and IL-10, and blood PGE2 levels were detected by Real-Time PCR and ELISA. Results: After LPS injection, the lungs showed significant edema and inflammatory cell infiltration. Compared with the normal group and the positive control group, the levels of TNF-α and PGE2 were increased by 145.1% (P<0.05) and 64.5% (P<0.05) respectively.Compared with the model group, TNF-α and PGE2 were reduced by 80.4% (P<0.05), 35.6% (P<0.05); but IL-4 and IL-10 levels were increased by 2.9 times (P<0.01) and 2.1 times (P<0.01) respectively in the PM groups. Conclusion: PM can significantly decrease the levels of TNF-α and PGE2 in lung tissue, attenuate the damage of alveolar cells from LPS, and protect lung tissue of mice with acute lung injury.