论文部分内容阅读
目的观察高体积分数氧(高氧)暴露时新生大鼠肺组织毛细血管内皮细胞中Src抑制的蛋白激酶C(PKC)的底物(SSeCKS)的表达及亚细胞定位的变化。方法出生12 h内大鼠128只按照随机数字表法分为4组:空气对照组、高氧暴露组、高氧暴露加蛋白激酶C抑制剂(R031-8220)组、高氧暴露加9 g/L盐水组。空气对照组置于空气中,余3组均置于常压高氧仓中,氧体积分数>950 mL/L,高氧暴露加R031-8220组自高氧暴露第1天起,腹腔注射5×10-2g/(kg.d)R031-8220,连续72 h;高氧暴露加9 g/L盐水组自高氧暴露第1天起,腹腔注射9 g/L盐水0.2 mL/(kg.d),连续72 h。于高氧暴露12、24、48和72 h时相点处死动物,在高氧暴露72 h行肺组织的病理学检查及肺湿/干质量比值(W/D)检测,应用免疫荧光双标法观察肺组织毛细血管内皮细胞SSeCKS的表达并通过IPP6.0软件对结果进行分析。采用SPSS13.0统计软件分析数据。结果高氧暴露组72 h时相点新生大鼠肺泡内可见大量红细胞,肺泡间隔增宽,粒细胞浸润;高氧暴露加R031-8220组新生大鼠肺组织病理改变较高氧暴露组明显减轻。与空气对照组比较,高氧暴露组W/D增加(t=9.223 P<0.01);与高氧暴露组比较,高氧暴露加R031-8220组W/D降低(t=5.050P<0.01)。72 h时相点其肺组织毛细血管内皮细胞SSeCKS的表达及亚细胞位置的变化:空气对照组SSeCKS主要分布在细胞膜;与空气对照组比较高氧暴露组SSeCKS表达增加,且有非常显著性差异(t=8.861 P<0.01),主要表达在细胞质中,高氧暴露加R031-8220组SSeCKS表达与高氧暴露组比较明显减少,有非常显著性差异(t=9.863 P<0.01),SSeCKS主要表达在细胞膜。结论新生大鼠肺组织毛细血管内皮细胞SSeCKS的变化参与高氧致新生大鼠肺组织毛细血管损伤的病理过程。
Objective To observe the changes of Src-suppressed protein kinase C (PKC) substrate (SSeCKS) expression and subcellular localization in pulmonary capillary endothelial cells of neonatal rats exposed to high volume fraction of oxygen (hyperoxia). Methods 128 rats were randomly divided into 4 groups: control group, hyperoxia exposure group, hyperoxia exposure plus RKC group (R031-8220) group, hyperoxia exposure plus 9 g / L saline group. Air control group placed in the air, the other three groups were placed in atmospheric oxygen chamber, oxygen volume fraction> 950 mL / L, high oxygen exposure plus R031-8220 group from the first day of exposure to oxygen, intraperitoneal injection of 5 × 10-2g / (kg · d) R031-8220 for 72 h; hyperoxia exposure plus 9 g / L saline group, intraperitoneal injection of 9 g / L saline 0.2 mL / (kg. d) for 72 h. The animals were sacrificed at 12, 24, 48 and 72 h after hyperoxia exposure. Pathological examination and W / D lung wet / dry weight ratio (W / D) were performed 72 h after hyperoxia exposure. Methods The expression of SSeCKS in pulmonary capillary endothelial cells was observed and the results were analyzed by IPP6.0 software. Using SPSS13.0 statistical software to analyze the data. Results In the hyperoxia exposure group, a large number of erythrocytes were found in the alveoli of neonatal rats at 72 h. The alveolar septum was widened and the infiltration of granulocytes was observed. The pathological changes in the lung tissue of neonatal rats exposed to hyperoxia exposure and R031-8220 group were significantly lower than those in the hyperoxia group . Compared with the air control group, the W / D of the hyperoxia group increased (t = 9.223 P <0.01), and the W / D of the group of high oxygen exposure plus R031-8220 decreased (t = 5.050 P <0.01) . The expression of SSeCKS and the changes of subcellular location in the pulmonary capillary endothelial cells at 72 h: the SSeCKS of the air control group mainly distributed in the cell membrane; the expression of SSeCKS in the hyperoxia exposed group increased compared with the air control group, and there was a very significant difference (t = 8.861 P <0.01). The expression of SSeCKS in R031-8220 group was significantly lower than that in hyperoxia group (t = 9.863 P <0.01) Expressed in the cell membrane. Conclusion The changes of SSeCKS in the pulmonary capillary endothelial cells of neonatal rats are involved in the pathological process of pulmonary injury induced by hyperoxia in neonatal rats.