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Effect of calcium-sensitive receptors on endothelial nitric oxide synthase and nitric oxide in neonatal mice with persistent pulmonary hypertension
LI Xiang, WU Bing-Xia, LI He, WANG Meng-Meng, MA Ke-Tao, GU Qiang
Chinese Journal of Contemporary Pediatrics ›› 2019, Vol. 21 ›› Issue (2) : 189-194.
PDF(2281 KB)
PDF(2281 KB)
Effect of calcium-sensitive receptors on endothelial nitric oxide synthase and nitric oxide in neonatal mice with persistent pulmonary hypertension
Objective To study the effect of calcium-sensitive receptors (CaSR) on the expression of endothelial nitric oxide synthase (eNOS) and the concentration of nitric oxide (NO) in a neonatal mouse model of persistent pulmonary hypertension (PPH). Methods Eighty neonatal C57BL/6 mice were randomly divided into control, PPH, agonist and antagonist groups. The control group was exposed to air, and the other three groups were exposed to 12% oxygen. The agonist and antagonist groups were intraperitoneally injected with a CaSR agonist (GdCl3 16 mg/kg) and a CaSR antagonist (NPS2390, 1 mg/kg), respectively, while the PPH and control groups were intraperitoneally injected with normal saline instead. All mice were treated for 14 days. Alveolar development and pulmonary vessels were assessed by hematoxylin-eosin staining. The protein and mRNA expression of eNOS and its localization in lung tissues were determined by Western blot, qRT-PCR and immunohistochemistry. The levels of brain natriuretic peptide (BNP) and NO in lung homogenate were determined using ELISA. Results Compared with the control group, the PPH and agonist groups showed significant increases in alveolar mean linear intercept, the percent wall thickness of pulmonary arterioles, right to left ventricular wall thickness ratio (RV/LV) and BNP concentration, but a significant reduction in radial alveolar count (P < 0.05). The antagonist group had significant improvements in all the above indices except RV/LV compared with the PPH and agonist groups (P < 0.05). Compared with those in the control group, the protein and mRNA expression of eNOS and NO concentration were significantly increased in the PPH group and increased more significantly in the agonist group, but were significantly reduced in the antagonist group (P < 0.05). Conclusions CaSR plays an important role in the development of PPH in neonatal mice, possibly by increasing eNOS expression and NO concentration.
Persistent pulmonary hypertension / Calcium-sensitive receptor / Endothelial nitric oxide synthase / Nitric oxide / Neonatal mice
[1] Nakwan N. The practical challenges of diagnosis and treatment options in persistent pulmonary hypertension of the newborn:a developing country's perspective[J]. Am J Perinatol, 2018, 35(14):1366-1375.
[2] Drummond WH. Neonatal pulmonary hypertension[J]. Equine Vet J, 1987, 19(3):169-171.
[3] Godo S, Sawada A, Saito H, et al. Disruption of physiological balance between nitric oxide and endothelium-dependent hyperpolarization impairs cardiovascular homeostasis in mice[J]. Arterioscler Thromb Vasc Biol, 2016, 36(1):97-107.
[4] 王萌萌, 李贺, 张方方, 等. 钙敏感受体在新生小鼠持续性肺动脉高压中的作用[J]. 中国当代儿科杂志, 2017, 19(2):208-214.
[5] 李贺, 谷强, 王萌萌, 等. 钙敏感受体对低氧诱导的持续性肺动脉高压小鼠肺动脉平滑肌细胞内钙离子浓度的影响[J]. 中国新生儿科杂志, 2018, 33(1):59-64.
[6] Ambalavanan N, Bulger A, Murphy-Ullrich J, et al. Endothelin-A receptor blockade prevents and partially reverses neonatal hypoxic pulmonary vascular remodeling[J]. Pediatr Res, 2005, 57(5 Pt 1):631-636.
[7] Knudsen L, Weibel ER, Gundersen HJ, et al. Assessment of air space size characteristics by intercept (chord) measurement:An accurate and efficient stereological approach[J]. J Appl Physiol (1985), 2010, 108(2):412-421.
[8] Cooney TP, Thurlbeck WM. The radial alveolar count method of emery and mithal:a reappraisal 1-postnatal lung growth[J]. Thorax, 1982, 37(8):572-579.
[9] Xu XF, Gu WZ, Wu XL, et al. Fetal pulmonary vascular remodeling in a rat model induced by hypoxia and indomethacin[J]. J Matern Fetal Neonatal Med, 2011, 24(1):172-182.
[10] Wedgwood S, Lakshminrusimha S, Schumacker PT, et al. Hypoxia inducible factor signaling and experimental persistent pulmonary hypertension of the newborn[J]. Front pharmacol, 2015, 6:47.
[11] Siragusa M, Fleming I. The eNOS signalosome and its link to endothelial dysfunction[J]. Pflugers Arch, 2016, 468(7):1125-1137.
[12] Rudic RD, Shesely EG, Maeda N, et al. Direct evidence for the importance of endothelium-derived nitric oxide in vascular remodeling[J]. J Clin Invest, 1998, 101(4):731-736.
[13] Kruzliak P, Maruyama J, Maruyama K. Role of nitric oxide in pathophysiology and treatment of pulmonary hypertension[J]. Vitam Horm, 2014, 96:407-424.
[14] Komai H, Naito Y, Aimi Y, et al. Nitric oxide synthase expression in lungs of pulmonary hypertensive patients with heart disease[J]. Cardiovasc Pathol, 2001, 10(1):29-32.
[15] 曹静, 朱艳萍, 李明霞. 缺氧诱导因子-1α及下游因子在缺氧性肺动脉高压新生大鼠肺内的表达研究[J]. 中国新生儿科杂志, 2014, 29(3):194-199.
[16] 桑葵, 周英, 李明霞. 缺氧诱导因子-1α及其调控因子在缺氧性肺动脉高压新生大鼠肺组织的蛋白质表达研究[J]. 中华儿科杂志, 2012, 50(12):919-924.
