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目的:探讨羊膜腔内注射肺表面活性物质(PS)对宫内感染致肺损伤胎兔的肺组织转化生长因子β1(TGF-β1)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,建立宫内感染模型。对照组和细菌组按胎龄又分为21、22、23、24、26 d 5个亚组,细菌+PS组分为24、26 d 2个亚组。细菌组宫内注射E.coli菌株,细菌+PS组宫内注射E.coli菌株的同时羊膜腔内注射PS200 mg/kg,对照组宫内注射生理盐水。模型建成后分别在各时相点取胎兔肺组织,免疫组织化学法检测肺组织内TGF-β1表达;RT-PCR检测TGF-β1mRNA表达;Western blotting法检测TGF-β1蛋白表达。结果:对照组TGF-β1蛋白在支气管上皮或血管内皮细胞及其周围的结缔组织有阳性表达,细菌组、细菌+PS组TGF-β1阳性蛋白表达量增加,与对照组比较,差异有统计学意义(P<0.05)。TGF-β1mRNA、蛋白表达在细菌感染后明显上调,3组比较,差异有显著统计学意义(F=631.109、183.980,P=0.000)。结论:宫内感染后胎肺TGF-β1高表达是肺损伤的高危因素。羊膜腔内注射PS可使TGF-β1表达下降,对减轻肺损伤、促进肺发育具有积极作用。
Objective: To investigate the effect of intra-amniotic injection of pulmonary surfactant (PS) on the expression of transforming growth factor-β1 (TGF-β1) in lung tissue of fetal rabbits with lung injury induced by intrauterine infection. Methods: Nineteen healthy adult Japanese white rabbits of childbearing age were selected and randomly divided into control group, bacteria group and bacteria + PS group after successful conception, and intrauterine infection model was established. The control group and the bacterial group were divided into 5 subgroups of 21, 22, 23, 24, and 26 d according to the gestational age, and the bacterial + PS components were divided into 2 subgroups of 24 and 26 d. E. coli strains were intraperitoneally injected into the bacterial group, PS200 mg / kg was intra-amniotic injected with E. coli strains in the bacteria + PS group, and intrauterine saline was injected into the control group. After the model was established, the lung tissue of fetus and rabbit were taken at each time point. The expression of TGF-β1 in lung tissue was detected by immunohistochemistry. The expression of TGF-β1 mRNA was detected by RT-PCR and the expression of TGF-β1 by Western blotting. Results: The TGF-β1 protein in the control group was positive in the bronchial epithelial or vascular endothelial cells and the surrounding connective tissue. The expression of TGF-β1-positive protein in the bacterial and bacterial + PS groups was increased compared with the control group Significance (P <0.05). TGF-β1 mRNA and protein expression were significantly up-regulated after bacterial infection. There was significant difference between the three groups (F = 631.109, 183.980, P = 0.000). Conclusion: High expression of TGF-β1 in fetal lung after intrauterine infection is a risk factor for lung injury. Intra-amniotic injection of PS can decrease the expression of TGF-β1, and have a positive effect on reducing lung injury and promoting lung development.