目的 。 探讨不同氧（O2）浓度环境对肺动脉平滑肌细胞（PASMC）表型的影响及肺动脉高压的形成机制 方法 采用酶解法分离培养大鼠原代PASMC，经鉴定后将稳定传代的PASMC分别置于正常O2含量（C组）及55%O2（H55组）、75%O2（H75组）、95%O2（H95组）的密闭容器培养6 h。分别采用实时荧光定量PCR法和Western blot法检测α-平滑肌肌动蛋白（α-SMA）、平滑肌22α蛋白（SM22α）、骨桥蛋白（OPN）、基质金属蛋白酶2（MMP-2）的mRNA和蛋白表达。 结果 H55组的α-SMA、SM22α、OPN、MMP-2的mRNA和蛋白相对表达量与C组差异均无统计学意义（P均>0.05）；与C组和H55组比较，H75组和H95组α-SMA、SM22α的mRNA和蛋白相对表达量均更低，OPN、MMP-2的mRNA和蛋白相对表达量均更高（P均<0.05）；与H75组相比，H95组MMP-2的mRNA相对表达量更高（P<0.05）。 结论 O2浓度75%以上环境可通过诱导PASMC表型转化，使PASMC获得较强分泌能力，形成肺动脉高压病理基础，并且这种诱导作用呈现O2浓度依赖性。 Objective To investigate the influence of varied oxygen (O2) concentration environments on the phenotypic transformation of pulmonary artery smooth muscle cells (PASMC) and the mechanism of pulmonary hypertension. Methods Primary rat PASMC were isolated and cultured through the process of enzymatic digestion. Following identification, the stable passaged PASMC were subjected to a 6-hour incubation in sealed containers with normal O2 content (group C) and relative O2 content comprising 55% (group H55), 75% (group H75), and 95% (group H95). mRNA and protein expression of α-Actin (α-SMA), smooth muscle 22α (SM22α), osteopontin (OPN), and matrix metalloproteinase-2 (MMP-2) were measured using real-time quantitative PCR and western blot analysis. Results The H55 group displayed no significant difference from the C group in terms of mRNA and relative protein expression levels for α-SMA, SM22α, OPN, and MMP-2 (all P>0.05). On the other hand, groups H75 and H95 exhibited a reduction in mRNA and relative protein expression of α-SMA and SM22α, along with an increase in mRNA and relative protein expression of OPN and MMP-2 when compared with both the C and H55 groups (allP<0.05). The H95 group showed a higher relative mRNA expression of MMP-2 as compared to the H75 group (P<0.05). Conclusions Oxygen concentration environments of 75% or higher can serve as the foundation for the pathogenesis of pulmonary hypertension, essentially by inducing a phenotypic transformation in PASMC towards adopting a robust secretory function. This induction is contingent upon the concentration of oxygen present.
Abstract
Objective To investigate the influence of varied oxygen (O2) concentration environments on the phenotypic transformation of pulmonary artery smooth muscle cells (PASMC) and the mechanism of pulmonary hypertension. Methods Primary rat PASMC were isolated and cultured through the process of enzymatic digestion. Following identification, the stable passaged PASMC were subjected to a 6-hour incubation in sealed containers with normal O2 content (group C) and relative O2 content comprising 55% (group H55), 75% (group H75), and 95% (group H95). mRNA and protein expression of α-Actin (α-SMA), smooth muscle 22α (SM22α), osteopontin (OPN), and matrix metalloproteinase-2 (MMP-2) were measured using real-time quantitative PCR and western blot analysis. Results The H55 group displayed no significant difference from the C group in terms of mRNA and relative protein expression levels for α-SMA, SM22α, OPN, and MMP-2 (all P>0.05). On the other hand, groups H75 and H95 exhibited a reduction in mRNA and relative protein expression of α-SMA and SM22α, along with an increase in mRNA and relative protein expression of OPN and MMP-2 when compared with both the C and H55 groups (allP<0.05). The H95 group showed a higher relative mRNA expression of MMP-2 as compared to the H75 group (P<0.05). Conclusions Oxygen concentration environments of 75% or higher can serve as the foundation for the pathogenesis of pulmonary hypertension, essentially by inducing a phenotypic transformation in PASMC towards adopting a robust secretory function. This induction is contingent upon the concentration of oxygen present.
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