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. 2019 Jun 6;44(2):535–548. doi: 10.3892/ijmm.2019.4233

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Copyright: © Yang et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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SPHK1 silencing alleviates ORMDL3 induced airway barrier dysfunction. (A) SPHK1 protein expression in si-SPHK1/si-ctrl 16HBE cells was detected by western blotting. (B) si-SPHK1/si-ctrl cells were treated with TNF-α (10 ng/ml) for 5 days and TEER was measured daily. (C) Claudin-18 and E-cadherin mRNA expression levels in si-SPHK1/si-ctrl and si-SPHK1+TNF-α/si-ctrl+TNF-α cells were detected by qRT-PCR. (D-a) Sphk1, (b) ORMDL3, (c) Claudin-18, (d) E-cadherin and (e) ERK1/2 protein expression levels in si-SPHK1/si-ctrl and si-SPHK1+TNF-α/si-ctrl+TNF-α cells were detected by western blotting. (E) ORMDL3/Vector cells were treated with DMS (10 nM) for 5 days and TEER was measured daily. (F) Claudin-18 and E-cadherin mRNA expression levels in ORMDL3/Vector and ORMDL3+DMS/Vector+DMS cells were detected by qRT-PCR. (G-a) Sphk1, (b) ORMDL3, (c) Claudin-18, (d) E-cadherin and (e) ERK1/2 protein expression levels in ORMDL3+DMS/Vector+DMS cells were detected by western blotting. Data are presented as the mean ± standard deviation of six independent experiments. ^**P<0.01 vs. si-ctrl group. ^#P<0.05, ^##P<0.01 and ^###P<0.001 vs. si-ctrl+TNF-a group. ^★P<0.05 vs. the Vector group. ^∆P<0.05, ^∆∆P<0.01 and ^∆∆∆P<0.001 vs. ORMDL3 group. E, epithelial; SPHK1, sphingosine kinase 1; ORMDL3, orosomucoid-like protein isoform 3; si, small interfering; ctrl, control; TNF, tumor necrosis factor; ERK, extracellular signal regulated kinase.

SPHK1 silencing alleviates ORMDL3 induced airway barrier dysfunction. (A) SPHK1 protein expression in si-SPHK1/si-ctrl 16HBE cells was detected by western blotting. (B) si-SPHK1/si-ctrl cells were treated with TNF-α (10 ng/ml) for 5 days and TEER was measured daily. (C) Claudin-18 and E-cadherin mRNA expression levels in si-SPHK1/si-ctrl and si-SPHK1+TNF-α/si-ctrl+TNF-α cells were detected by qRT-PCR. (D-a) Sphk1, (b) ORMDL3, (c) Claudin-18, (d) E-cadherin and (e) ERK1/2 protein expression levels in si-SPHK1/si-ctrl and si-SPHK1+TNF-α/si-ctrl+TNF-α cells were detected by western blotting. (E) ORMDL3/Vector cells were treated with DMS (10 nM) for 5 days and TEER was measured daily. (F) Claudin-18 and E-cadherin mRNA expression levels in ORMDL3/Vector and ORMDL3+DMS/Vector+DMS cells were detected by qRT-PCR. (G-a) Sphk1, (b) ORMDL3, (c) Claudin-18, (d) E-cadherin and (e) ERK1/2 protein expression levels in ORMDL3+DMS/Vector+DMS cells were detected by western blotting. Data are presented as the mean ± standard deviation of six independent experiments. ^**P<0.01 vs. si-ctrl group. ^#P<0.05, ^##P<0.01 and ^###P<0.001 vs. si-ctrl+TNF-a group. ^★P<0.05 vs. the Vector group. ^∆P<0.05, ^∆∆P<0.01 and ^∆∆∆P<0.001 vs. ORMDL3 group. E, epithelial; SPHK1, sphingosine kinase 1; ORMDL3, orosomucoid-like protein isoform 3; si, small interfering; ctrl, control; TNF, tumor necrosis factor; ERK, extracellular signal regulated kinase.