Abstract：【Objective】To investigate the role and mechanism of NR2D subunit of NMDA receptor in hyperoxia-induced lung injury of neonatal rats. 【Methods】Animal experiments were performed in a total of 40 newborn SD rats (within 12 hours of birth) . The rats were randomly divided into the control group (Con), 1 day after hyperoxia exposure group, 7 days after hyperoxia exposure group and 14 days after hyperoxia exposure group, with 10 rats in each group. Neonatal rats were exposed to hyperoxia (oxygen content ≥95%) for 3 days to establish lung injury model induced by hyperoxia. The neonatal rats in the hyperoxia group were sacrificed with the control group mice at 1, 7, 14 days after the establishment of lung injury model. Hematoxylin-eosin staining (HE) and Masson staining were used to observe the pathological changes of lung in rats of each group; then Holfbauer score was performed. The content of hydroxyproline (HYP) in lung tissue was measured by a hydroxyproline detection kit. The expression of NR2D subunit was detected by Western blot. In vitro experiments, rat alveolar epithelial cell line RLE-6TN was divided into six groups: the blank control group, the model group, NR2D overexpression plasmid group, NR2D empty plasmid group, NR2D siRNA group and NR2D siRNA negative control group. Cells in the model group and the transfected groups were exposed to hyperoxia to establish the lung cell injury status. After transfection of plasmid and siRNA, cell proliferation was detected by CKK-8 assay; levels of cytokines TNF-α, IL-6 and TGF-β secreted by cells were measured by enzyme-linked immunosorbent assay (ELISA); expressions of NR2D subunit, E-cadherin and Vimentin proteins were shown by Western blot. 【Results】Animal experiment results showed that, the Holfbauer score, HYP content and expression level of NR2D subunit of neonatal rats in the hyperoxia exposure group were higher than those in the control group and the trend of increase was in a time-dependent manner (P<0.05). Cell experiment results showed that, compared to the control group, cell proliferation and E-cadherin protein expression in the model group decreased (P<0.05), while the expressions of NR2D subunit, TNF-α , IL-6 , TGF-β and Vimentin increased (P<0.05). Compared to the model group, cell proliferation and E-cadherin protein expression in NR2D overexpression group decreased (P<0.05), while the NR2D subunit expression and protein expressions of TNF-α, IL-6, TGF-β and Vimentin increased (P<0.05). On the other hand, relative cell proliferation rate and E-cadherin protein expression in NR2D siRNA group were increased (P<0.05), while the NR2D subunit expression and protein expressions of TNF-α, IL-6, TGF-β and Vimentin were decreased (P<0.05). There were no significant changes of above indexes in both NR2D empty plasmid group and NR2D siRNA negative control group (P>0.05). 【Conclusion】NMDA receptor NR2D subunit plays an important role in the occurrence and development of lung injury induced by hyperoxia in neonatal rats. Downregulation of its expression protects alveolar epithelial cells from hyperoxia injury, possibly by reducing levels of cytokines TNF-α, IL-6 and TGF-β.
张喜, 李林瑞, 段效军, 陈艳萍, 王铭杰. NMDA受体NR2D亚基在新生大鼠高氧性肺损伤中的作用及其机制[J]. 医学临床研究, 2019, 36(11): 2095-2100.
ZHANG Xi, LI Lin-rui, DUAN Xiao-jun, et al. Investigation on the Role and Mechanism of NR2D Subunit of NMDA Receptor in Hyperoxia-induced Lung Injury of Neonatal SD Rats. JOURNAL OF CLINICAL RESEARCH, 2019, 36(11): 2095-2100.
[1] 张媛,程怀平,包天平, 等. 高氧诱导肺损伤新生小鼠肺组织中长链非编码RNA NANCI的表达及对NKX2.1的调控作用[J].中国当代儿科杂志,2017, 19(2):215-221. [2] Syed M, Das P, Pawar A,et al. Hyperoxia causes miR-34a-mediated injury via angiopoietin-1 in neonatal lungs[J].Nat Commun,2017,8(1):1173. [3] Murrough JW, Abdallah CG, Mathew SJ. Targeting glutamate signalling in depression: progress and prospects[J].Nat Rev Drug Discov,2017,16(7):472-486. [4] Liao Z, Zhou X, Luo Z, et al. N-Methyl-D-aspartate receptor excessive activation inhibited fetal rat lung development in vivo and in vitro[J].Biomed Res Int,2016, 2016:5843981. [5] Wang YR, Yue SJ, Luo ZQ, et al. N-methyl-D-aspartate receptor activation mediates lung fibroblast proliferation and differentiation in hyperoxia-induced chronic lung disease in newborn rats[J].Respir Res,2016, 17(1):136. [6] 马玲,刘伟,冯丹丹,等NMDA受体阻断剂美金刚胺对小鼠内毒素急性肺损伤的保护作用[J].中南大学学报(医学版), 2014, 39(1):12-16. [7] 徐惠琴, 杜延茹, 沈晶赞, 等. NMDA受体NR2A、NR2B亚基在戊四氮诱导慢性癫痫大鼠中的作用[J].温州医科大学学报, 2018, 48(12): 866-870. [8] Cerpa W, Gambrill A, Inestrosa NC, et al. Regulation of NMDA-receptor synaptic transmission by Wnt signaling[J].J Cogn Neurosci,2016, 31(26):9466-9471. [9] Cui H, He J, Chen H, et al. Erythropoietin attenuates hyperoxia-induced lung injury by upregulating epidermal growth factor-like domain 7 in newborn rats[J].Biomed Rep,2017, 6(1):32-38. [10] Bukowczan J,Warzecha Z,Ceranowicz P,et al.Pretreatment with obestatin reduces the severity of ischemia/reperfusion-induced acute pancreatitis in rats[J].Eur J Pharmacol,2015,760 ( 5) :113-121. [11] 杨文彬,赵小青,梁冉,等.小分子RNA干扰雷帕霉素靶蛋白基因表达对百草枯致大鼠肺纤维化的影响[J].中华危重病急救医学, 2017, 29(9):830-835. [12] Lu HY,Chen XQ,Tang W,et al. GRP78 silencing enhances hyperoxia-induced alveolar epithelial cell apoptosis via CHOP pathway[J].Mol Med Rep,2017, 16(2):1493-1501. [13] Hummler JK, Dapaah-Siakwan F, Vaidya R, et al. Inhibition of Rac1 signaling downregulates inflammasome activation and attenuates lung injury in neonatal rats exposed to hyperoxia[J].Neonatology,2017, 111(3):280-288. [14] Ito M, Nagano N, Arai Y, et al. Genetic ablation of Bach1 gene enhances recovery from hyperoxic lung injury in newborn mice via transient upregulation of inflammatory genes[J].Pediatr Res,2017, 81(6):926-931. [15] Bai W, Zhu WL, Ning YL, et al. Dramatic increases in blood glutamate concentrations are closely related to traumatic brain injury-induced acute lung injury[J].Sci Rep,2017, 7(1):5380. [16] Maillet I, Perche O, P ris A, et al. Glufosinate aerogenic exposure induces glutamate and IL-1 receptors dependent lung inflammation[J].Clin Sci(Lond), 2016, 130(21):1939-1954. [17] Shi T S, Hao JX,Wiesenfeld-Hallin Z, et al. Gabapentin and NMDA receptor antagonists interacts synergistically to alleviate allodynia in two rat models of neuropathic pain[J].Scand J Pain,2018,18(4): 687-693. [18] Fu H, Zhang T, Huang R, et al. Calcitonin gene-related peptide protects type II alveolar epithelial cells from hyperoxia-induced DNA damage and cell death[J].Exp Ther Med,2017, 13(4):1279-1284.
2019, Vol. 36 
