目的 探讨2-甲氧基雌二醇（2-methoxyestradiol, 2ME）在新生大鼠缺氧性肺动脉高压中的保护作用。 方法 96只Wistar新生大鼠随机分为常氧组、缺氧组和缺氧+2ME组，每组随机分为3 d、7 d、14 d、21 d亚组，每个亚组8只。缺氧组和缺氧+2ME组分别予以每日皮下注射生理盐水和2ME（剂量240 μg/kg），常氧组在常氧环境下饲养，每日皮下注射生理盐水。直接测压法测量右心室收缩压（right ventricular systolic pressure, RVSP）；苏木精-伊红染色观察肺血管形态，计算肺血管重塑指标肺小动脉中层血管壁厚度占血管外径的百分比（MT%）和肺小动脉中层截面积占血管总截面积的百分比（MA%）；免疫组化法检测肺组织中缺氧诱导因子-1α（hypoxia-inducible factor-1α, HIF-1α）、增殖细胞核抗原（proliferating cell nuclear antigen, PCNA）蛋白表达水平；实时荧光定量聚合酶链反应检测HIF-1α、PCNA mRNA表达水平。 结果 与常氧组比较，缺氧3、7、14、21 d时缺氧组、缺氧+2ME组RVSP升高，HIF-1α、PCNA蛋白和mRNA表达水平上调（P<0.05），缺氧7、14、21 d时缺氧组MT%、MA%增加（P<0.05）；与缺氧组比较，缺氧3、7、14、21 d时缺氧+2ME组RVSP降低，HIF-1α、PCNA蛋白和PCNA mRNA表达水平下调（P<0.05），缺氧7、14、21 d时缺氧+2ME组MT%、MA%减少（P<0.05）。 结论 2ME可能通过抑制HIF-1α、PCNA表达，减轻肺血管重塑，对新生大鼠缺氧性肺动脉高压发挥保护作用。
Abstract
Objective To investigate the protective effects of 2-methoxyestradiol (2ME) against hypoxic pulmonary hypertension (HPH) in neonatal rats. Methods Ninety-six Wistar neonatal rats were randomly divided into a normoxia group, a hypoxia group, and a hypoxia + 2ME group, with each group further subdivided into 3-day, 7-day, 14-day, and 21-day subgroups, containing eight rats each. The hypoxia and hypoxia + 2ME groups received daily subcutaneous injections of saline and 2ME (240 μg/kg), respectively, while the normoxia group was raised in a normoxic environment with daily saline injections. Right ventricular systolic pressure (RVSP) was measured using the direct pressure method. Pulmonary vascular morphology was assessed using hematoxylin and eosin staining, with metrics including the percentage of medial thickness of small pulmonary arteries relative to the external diameter (MT%) and the cross-sectional area of the media of small pulmonary arteries relative to the total cross-sectional area (MA%). Immunohistochemistry was used to detect the expression levels of hypoxia-inducible factor-1α (HIF-1α) and proliferating cell nuclear antigen (PCNA) proteins, while real-time quantitative PCR was used to to assess HIF-1α and PCNA mRNA levels. Results Compared to the normoxia group, the hypoxia and hypoxia + 2ME groups showed increased RVSP and upregulated HIF-1α and PCNA protein and mRNA expression levels at 3, 7, 14, and 21 days after hypoxia (P<0.05). Furthermore, at 7, 14, and 21 days after hypoxia, the hypoxia group showed increased MT% and MA% (P<0.05). In comparison to the hypoxia group, the hypoxia + 2ME group exhibited reduced RVSP and downregulated HIF-1α and PCNA protein and mRNA expression levels, along with decreased MT% and MA% at 7, 14, and 21 days after hypoxia (P<0.05). Conclusions 2ME may protect against HPH in neonatal rats by inhibiting the expression of HIF-1α and PCNA and reducing pulmonary vascular remodeling. Citation:Chinese Journal of Contemporary Pediatrics, 2024, 26(7): 757-764
关键词
缺氧性肺动脉高压 /
2-甲氧基雌二醇 /
缺氧诱导因子-1α /
新生大鼠
Key words
Hypoxic pulmonary hypertension /
2-Methoxyestradiol /
Hypoxia-inducible factor-1α /
Neonatal rat
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