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目的观察大鼠吸入1.5%异氟烷不同时间后肺表面活性蛋白-A(SP-A)的含量及其基因表达的改变情况。方法雄性Wister大鼠40只,随机分为5组,每组8只。正常对照组(Con),异氟烷2h组、4h组和8h组分别为吸入40%O2+1.5%异氟醚2、4和8h组;停药2h组(10h组)吸入40%O2+1.5%异氟醚8h后再吸入40%O22h。各组于实验结束后取出大鼠迅速处死,检测支气管肺泡灌洗液(BALF)中表面活性蛋白A(SP-A)的含量(WesternBlot法);肺组织内SP-A含量(WesternBlot法)以及SP-AmRNA的表达(RT-PCR法)。结果吸入异氟烷2h组BALF中SP-A的含量与对照组比较无明显变化,4h、8h组均较对照组显著下降(P<0.01),10h组未见明显恢复(P<0.01)。肺组织内SP-A的含量在2h组无明显改变,但4h、8h组明显下降(P<0.01),10h组有所恢复,但仍低于对照组(P<0.05)。吸入异氟烷的各组SP-AmRNA的表达均有降低趋势(P<0.05),停药2h后(10h)明显恢复。结论吸入1.5%异氟烷2h以上可降低肺组织SP-A的基因表达,继而SP-A的合成与分泌减少;但停止吸入2h后,SP-A基因表达可恢复至正常水平,而其合成与分泌功能仍未能完全恢复。
Objective To investigate the changes of lung surfactant protein-A (SP-A) and its gene expression after 1.5% isoflurane inhalation in rats. Methods Forty male Wister rats were randomly divided into 5 groups with 8 rats in each group. In the control group (Con), isoflurane 2h, 4h and 8h groups were inhaled 40% O2 + 1.5% isoflurane 2,4 and 8h respectively; inhaled 40% O2 + 1.5% isoflurane 8h and then inhaled 40% O22h. Rats in each group were sacrificed immediately after the completion of the experiment, and the content of surfactant protein A (SP-A) in bronchoalveolar lavage fluid (BALF) was determined by Western Blotting method; the content of SP-A in lung tissue (Western Blot method) SP-AmRNA expression (RT-PCR method). Results Compared with the control group, the content of SP-A in BALF of inhaled isoflurane 2h group was significantly lower than that of the control group (P <0.01), and no significant recovery was found in 10h group (P <0.01). The content of SP-A in lung tissue did not change significantly in 2h group, but significantly decreased in 4h, 8h (P <0.01), and recovered in 10h group, but still lower than that in control group (P <0.05). The expression of SP-AmRNA in each group inhaled isoflurane decreased (P <0.05), after 2h withdrawal (10h) significantly recovered. Conclusions Inhalation of 1.5% isoflurane for 2 h can reduce the expression of SP-A gene in lung tissue and decrease the synthesis and secretion of SP-A. However, the expression of SP-A gene returned to normal level after 2 h of inhalation, And secretory function still failed to fully recover.