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目的探讨钙敏感受体(CaSR)及相关分子在持续肺动脉高压(PPH)新生大鼠肺组织中的表达及在PPH发病机制中的作用。方法选取6只Sprague-Dawley(SD)孕鼠,孕第19天时随机分为PPH组和对照组,每组3只。PPH组孕鼠置于12%氧浓度的缺氧箱中并腹腔内注射吲哚美辛0.5 mg/kg,2次/天,连续3天,建立胎鼠肺动脉高压模型;对照组孕鼠暴露在空气中并腹腔内注射等量生理盐水。两组孕鼠均于孕第22天行剖宫产,对照组、PPH组分别获取新生大鼠26、28只,两组各随机抽取15只仔鼠用于后续实验。采用RT-PCR技术检测新生大鼠肺组织CaSR mRNA及Ⅱ型11β羟基类固醇脱氢酶(11β-HSD2)mRNA表达,采用蛋白质印迹法检测新生大鼠肺组织CaSR蛋白和一氧化氮(NO)含量。结果与对照组比较,PPH组新生大鼠肺组织中CaSR mRNA[(0.592±0.052)比(0.408±0.017)]及蛋白表达[(0.717±0.027)比(0.445±0.011)]升高,NO含量[(1.50±0.20)μmol/g比(0.37±0.16)μmol/g]升高,11β-HSD2 mRNA[(0.413±0.073)比(0.978±0.079)]表达降低,差异均有统计学意义(P<0.05)。结论 CaSR及相关分子在新生大鼠PPH发病中可能发挥重要作用。
Objective To investigate the expression of calcium sensitive receptor (CaSR) and related molecules in the lung tissue of neonatal rats with persistent pulmonary hypertension (PPH) and its role in the pathogenesis of PPH. Methods Six pregnant rats of Sprague-Dawley (SD) were selected and randomly divided into PPH group and control group on the 19th day of pregnancy, with 3 rats in each group. Pregnant rats in PPH group were placed in anoxia chamber with oxygen concentration of 12% and intraperitoneally injected with indomethacin 0.5 mg / kg twice daily for 3 days to establish a model of fetal pulmonary hypertension. Rats in control group were exposed to In the air and intraperitoneal injection of the same amount of saline. Two groups of pregnant rats were performed cesarean section on the 22nd day of pregnancy, while 26 and 28 neonatal rats were obtained from the control and PPH groups. 15 offspring were randomly selected for follow-up study in both groups. The expression of CaSR mRNA and 11β-hydroxysteroid dehydrogenase (11β-HSD2) mRNA in the lungs of neonatal rats were detected by RT-PCR. CaSR protein and NO content in the lungs of neonatal rats were detected by Western blotting . Results Compared with the control group, the levels of CaSR mRNA [(0.592 ± 0.052) (0.408 ± 0.017)] and protein expression in the lung tissue of PPH group [(0.717 ± 0.027) vs (0.445 ± 0.011)], NO content (1.50 ± 0.20) μmol / g (0.37 ± 0.16) μmol / g], while the expression of 11β-HSD2 mRNA (0.413 ± 0.073 vs 0.978 ± 0.079) <0.05). Conclusion CaSR and related molecules may play an important role in the pathogenesis of PPH in neonatal rats.