IL-6Ra in Smooth Muscle Cells Protects against Schistosoma- and Hypoxia-induced Pulmonary Hypertension.
IL-6Ra in Smooth Muscle Cells Protects against Schistosoma- and Hypoxia-induced Pulmonary Hypertension.
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平滑肌细胞中的 IL-6Ra 可预防血吸虫和缺氧引起的肺动脉高压。
DOI：
10.1165/rcmb.2018-0277le
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发表时间：
2019
影响因子：
6.4
通讯作者：
Graham,BrianB
中科院分区：
文献类型：
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作者：
Mickael,Claudia;Kumar,Rahul;Hernandez-Saavedra,Daniel;Kassa,Biruk;Sanders,Linda;Koyanagi,Dan;Gu,Sue;Lee,MichaelH;Tuder,RubinM;Graham,BrianB
方法购买Il6ra flox/flox和Myh11-CreER T2小鼠（Jackson实验室），杂交生成Il6ra flox/flox；Myh11-CreER T2小鼠。所有动物实验均经科罗拉多大学机构动物护理和使用委员会批准。详细信息请参见数据补充。结果处理Il6ra /flox；给Myh11-CreER T2小鼠注射他莫昔芬，并通过qPCR对在平滑肌选择性培养基上生长的解剖和消化的肺血管获得的平滑肌细胞进行定量缺失效率（> 90%的细胞为α-SMA+）。我们发现，平均有59%的Il6ra DNA被删除。我们挑战了他莫昔芬处理的Il6ra flox/flox；Myh11-CreER T2小鼠患有血吸虫病或缺氧，并发现通过右心室收缩压（RVSP）测量的PH严重程度与单独给玉米油后的任何一种刺激相比，任何一种刺激都更差（图1A）。我们通过观察他莫昔芬治疗小鼠肺血管介质厚度的增加，证实了他莫昔芬治疗小鼠更严重的PH值，这是通过免疫染色评估的α-平滑肌肌动蛋白（α-SMA）（图1B）或平滑肌球蛋白重链（数据补充中图E1）。
MethodsWe purchased Il6ra flox/flox and Myh11-CreER T2 mice (The Jackson Laboratory), and crossed the lines to generate Il6ra flox/flox; Myh11-CreER T2 mice. All animal experiments were approved by the University of Colorado Institutional Animal Care and Use Committee. For further details, see the data supplement.ResultsWe treated Il6ra flox/flox; Myh11-CreER T2 mice with tamoxifen and quantified the deletion efficiency by qPCR of smooth muscle cells obtained from dissected and digested pulmonary vessels grown on smooth muscle–selective media (> 90% of the cells were α-SMA+). We found that on average, 59% of the Il6ra DNA was deleted. We challenged tamoxifen-treated Il6ra flox/flox; Myh11-CreER T2 mice with either Schistosoma or hypoxia, and found that the PH severity as measured by right ventricle systolic pressure (RVSP) was worse with either stimuli as compared with either stimuli after corn oil administration alone (Figure 1A). We corroborated the finding of more severe PH in tamoxifen-treated mice by observing increased pulmonary vascular media thickness as assessed by immunostaining for either α-smooth muscle actin (α-SMA)(Figure 1B) or smooth muscle myosin heavy chain (Figure E1 in the data supplement) in Schistosoma-challenged mice that had received tamoxifen.