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目的探讨外源性肺表面活性物质(PS)对宫内感染所致肺损伤胎兔肺组织IL-8表达的影响。方法选取健康成年育龄日本大耳白兔19只,成功受孕后随机分为对照组、细菌组、细菌+PS组。建立宫内感染致肺损伤胎兔模型,对照组和细菌组按胎龄又分为21d、22d、23d、24d、26d5个亚组;细菌+PS组分为24d、26d2个亚组。细菌组宫内注射E.coli菌株,细菌+PS组宫内注射E.coli菌株,同时胎兔羊膜腔内注射PS200mg·kg-1,对照组宫内注射9g.L-1盐水。模型建成后分别在各时间点对孕兔行剖宫产,杀取胎兔后取肺组织,以鼠抗兔IL-8单克隆抗体为一抗,用免疫组织化学方法检测其肺组织内IL-8表达;用反转录-PCR检测其IL-8mRNA表达;用Westernblotting方法检测其IL-8蛋白表达;应用SPSS13.0软件进行统计学处理。结果细菌感染后胎兔肺组织IL-8mRNA、IL-8蛋白表达增高,与对照组比较差异有统计学意义(Pa=0.000)。免疫组织化学结果显示对照组支气管黏膜上皮细胞、平滑肌细胞、血管内皮细胞及巨噬细胞胞质内可见阳性表达,细菌组和细菌+PS组表达增强,在肺泡上皮细胞胞质也见明显表达,不同组间比较差异有统计学意义(F=79.785,P=0.000)。注射PS后胎兔肺组织IL-8mRNA、IL-8蛋白表达低于细菌组,但仍高于对照组。结论在感染环境中,羊膜腔内注射PS对减轻肺损伤、促进肺发育具有积极作用。
Objective To investigate the effect of exogenous pulmonary surfactant (PS) on the expression of IL-8 in lung tissue of fetal rabbits with lung injury caused by intrauterine infection. Methods Nineteen Japanese white rabbits of childbearing age were selected and randomly divided into control group, bacteria group and bacteria + PS group after successful conception. The model of fetal lung injury caused by intrauterine infection was established. The control group and bacteria group were divided into 21 subgroups at 21 days, 22 days, 23 days, 24 days and 26 days respectively. Bacteria + PS components were divided into 24 subgroups and 26 days subgroups. E. coli strains were intrauterinely injected into the bacterial group, and E. coli strains were intrauterinely injected into the bacteria + PS group. At the same time, intraperitoneal injection of PS200mg · kg-1 was given to the fetus and 9g.L-1 saline was injected into the control group. After the model was established, cesarean section was performed on the pregnant rabbits at each time point, and the lung tissue was taken after the fetus was killed. The anti-rabbit IL-8 monoclonal antibody was used as the primary antibody. The level of IL -8 expression. The expression of IL-8 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). The protein expression of IL-8 was detected by Western blotting. SPSS13.0 software was used for statistical analysis. Results The levels of IL-8 mRNA and IL-8 protein in lung tissue of rabbits after bacterial infection were significantly higher than those in control group (Pa = 0.000). Immunohistochemical results showed that in the control group, bronchial epithelial cells, smooth muscle cells, vascular endothelial cells and macrophage cytoplasm showed positive expression, bacterial group and bacteria + PS group increased expression in the alveolar epithelial cells were also seen in the cytoplasm, The difference between different groups was statistically significant (F = 79.785, P = 0.000). The expression of IL-8mRNA and IL-8 in pulmonary tissue of rabbits injected with PS was lower than that of the bacteria group, but still higher than that of the control group. Conclusion In the infected environment, amniotic injection of PS has a positive effect on reducing lung injury and promoting lung development.