实验性先天性膈疝中肺动脉内皮细胞与平滑肌细胞相互作用的改变
Altered pulmonary artery endothelial-smooth muscle cell interactions in experimental congenital diaphragmatic hernia.
作者信息
Acker Shannon N, Seedorf Gregory J, Abman Steven H, Nozik-Grayck Eva, Kuhn Katherine, Partrick David A, Gien Jason
机构信息
1] Department of Surgery, University of Colorado School of Medicine, Aurora, Colorado [2] Department of Pediatrics, The Pediatric Heart Lung Center, Children's Hospital Colorado, University of Colorado School of Medicine, Aurora, Colorado.
1] Department of Pediatrics, The Pediatric Heart Lung Center, Children's Hospital Colorado, University of Colorado School of Medicine, Aurora, Colorado [2] Department of Pediatrics, University of Colorado School of Medicine, Aurora, Colorado.
出版信息
Pediatr Res. 2015 Apr;77(4):511-9. doi: 10.1038/pr.2015.13. Epub 2015 Jan 12.
BACKGROUND
Pulmonary hypertension (PH) secondary to vascular remodeling contributes to poor outcomes in congenital diaphragmatic hernia (CDH), however mechanisms responsible are unknown. We hypothesized that pulmonary artery endothelial cell (PAEC) dysfunction contributes to smooth muscle cell (SMC) hyperplasia in experimental CDH.
METHODS
PAEC and SMC were isolated from fetal sheep with experimental CDH and controls. SMC growth was assessed alone and with SOD plus catalase and during coculture with control or CDH PAEC with and without ET-1 siRNA transfection. ET-1 protein was measured in PAEC and SMC lysates and supernatant. ROS production was measured in normal and CDH PAECs with and without ET-1 siRNA. PAEC growth and tube formation were measured with SOD plus catalase.
RESULTS
CDH SMC growth was decreased and increased with coculture with CDH PAEC more than control PAEC. Treatment of CDH PAEC with SOD plus catalase or ET-1 siRNA prevented the increase in SMC growth seen with coculture. ET-1 protein was increased in CDH PAEC and SMC. ROS production was increased in CDH PAEC and decreased with ET-1 SiRNA. SOD plus catalase restored CDH PAEC growth and tube formation.
CONCLUSION
PAEC dysfunction in experimental CDH increases SMC proliferation via ET-1 induced ROS production by PAEC.
背景
血管重塑继发的肺动脉高压（PH）导致先天性膈疝（CDH）预后不良，但其相关机制尚不清楚。我们推测在实验性CDH中，肺动脉内皮细胞（PAEC）功能障碍导致平滑肌细胞（SMC）增生。
方法
从患有实验性CDH的胎羊和对照胎羊中分离出PAEC和SMC。单独评估SMC生长情况，以及在与超氧化物歧化酶（SOD）加过氧化氢酶共同培养时，以及在与对照或CDH PAEC共培养且有或无内皮素-1（ET-1）小干扰RNA转染的情况下评估SMC生长情况。检测PAEC和SMC裂解物及上清液中的ET-1蛋白。在有或无ET-1小干扰RNA的情况下，检测正常和CDH PAEC中的活性氧（ROS）生成情况。用SOD加过氧化氢酶检测PAEC的生长和管腔形成情况。
结果
与对照PAEC相比，CDH SMC与CDH PAEC共培养时生长减少且增加。用SOD加过氧化氢酶或ET-1小干扰RNA处理CDH PAEC可阻止共培养时SMC生长的增加。CDH PAEC和SMC中的ET-1蛋白增加。CDH PAEC中的ROS生成增加，而ET-1小干扰RNA可使其减少。SOD加过氧化氢酶恢复了CDH PAEC的生长和管腔形成。
结论
实验性CDH中的PAEC功能障碍通过PAEC产生的ET-1诱导ROS生成增加SMC增殖。
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