Differential regulation of the genes encoding platelet-derived growth factor receptor and its ligand in rat lung during microvascular and alveolar wall remodeling in hyperoxia.
Differential regulation of the genes encoding platelet-derived growth factor receptor and its ligand in rat lung during microvascular and alveolar wall remodeling in hyperoxia.
复制标题
高氧条件下微血管和肺泡壁重塑过程中大鼠肺中编码血小板衍生生长因子受体及其配体的基因的差异调节。
DOI：
10.1165/ajrcmb/7.3.278
复制
发表时间：
1992
影响因子：
6.4
通讯作者：
Jones,R
中科院分区：
文献类型：
--
作者：
Powell,PP;Wang,CC;Jones,R
Materials and MethodsAnimal Model Viral antibody-free male Sprague-Dawley rats (175 to 185 g) were obtained from Charles River Laboratories (Kingston, NY). Animals were maintained on a 12-h light/dark cycle at 22 C. Treated rats breathed 87% oxygen at normobaric pressure (18, 19). Control rats were maintained in air. Rats in hyperoxia gained less weight than their age-matched controls. At the time of tissue collection, the animals received sodium pentobarbitol (30 mg/kg, ip).RNA Isolation Forty-four rats (n= 4 at each time point) were exposed to 87% oxygen for time periods of 2, 4, 6, 12, or 18 h or 1, 3, 7, 14, 21, or 28 days. Control rats were killed on days 1 and 28 (n= 4 at each time point). The early time points (hours) were necessary to establish a response for the receptors. The later times (days) were chosen to coincide with ['Hltbymidine labeling on days 1, 7, and 28 and to give extra time points between. Day 3 was chosen to examine any change in the time immediately before the proliferative response occurring in the microvessels (4), and day 14 as the midpoint between days 7 and 28. Lungs and heart were removed quickly, placed in a sterile petri dish, and thin strips of peripheral tissue (2 mm thick) dissected from the single lobed left lung and from the cardiac and diaphragmatic lobes of the right lung. Tissue was immediately dropped into liquid nitrogen and stored at-70 C. Frozen tissue was homogenized in 4 M guanidinium isothiocyanate (Brinkman polytron PT 3000). The homogenate was centrifuged briefly (1,000 X g) to sediment fibrous material. The clear supernatant was layered onto a cushion of 5.7 M cesium chloride. Total RNA was purified by density gradient centrifugation at 120,000 x g. The RNA pellet was resuspended in DEPC-treated water, precipitated with ethanol, and stored at-70 C.
