Effect of the Notch4/Dll4 signaling pathway in early gestational intrauterine infection on lung development

Zhan,Canyang; Sun,Yi; Pan,Jiarong; Chen,Lihua; Yuan,Tianming

doi:10.3892/etm.2021.10404

Effect of the Notch4/Dll4 signaling pathway in early gestational intrauterine infection on lung development

Authors:
- Canyang Zhan
- Yi Sun
- Jiarong Pan
- Lihua Chen
- Tianming Yuan
View Affiliations

Affiliations: Department of Neonatology, Children's Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, P.R. China
Published online on: July 8, 2021 https://doi.org/10.3892/etm.2021.10404
Article Number: 972
Copyright: © Zhan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Intrauterine infection is an important risk factor for bronchopulmonary dysplasia (BPD). BPD is characterized by arrested lung alveolarization and impaired pulmonary vascularization. The Notch4 signaling pathway is a key regulator of vascular remodeling and angiogenesis. Therefore, the presents study investigated the expression of Notch4, delta‑like canonical Notch ligand 4 (Dll4) and related factors in an in vivo rat model and in rat pulmonary microvascular endothelial cells (PMVECs) in vitro, to study the mechanisms by which intrauterine infection affects rat lung development. A rat model of intrauterine infection was established by endocervical inoculation with Escherichia scoli on embryonic day 15. The date of birth was counted as postnatal day 0 (P0). Then, the lung tissues were collected from pups at days P3‑P14. The expression of Notch4, Dll4 and related factors was measured by reverse transcription‑quantitative PCR and western blotting. In addition, the γ‑secretase inhibitor DAPT was used to examine the effect of Notch4 signaling on PMVECs. Intrauterine E. coli infection impaired normal lung development, as indicated by decreased microvessel density, fewer alveoli, fewer secondary septa, and larger alveoli compared with the control group. Furthermore, Notch4, Dll4 and NF‑κB levels were significantly increased in the E. coli‑infected group at P3 compared with the control group. Similarly, the mRNA expression levels of fetal liver kinase 1 (Flk‑1, a VEGF receptor) were significantly increased in the E. coli‑infected group at P3 and P7. In PMVECs, the inhibition of Notch4 signaling contributed to decreases in lipopolysaccharide (LPS)‑induced expression of VEGF and its receptors. Furthermore, the inhibition of Notch4/Dll4 signaling accelerated cell proliferation and decreased the apoptosis rate of LPS‑induced PMVECs. LPS‑induced NF‑κB expression in PMVECs was also attenuated by the Notch4/Dll4 inhibitor. In conclusion, intrauterine E. coli infection impaired normal lung development, possibly through Notch4/Dll4 signaling and effects on VEGF and its receptors.

View Figures

View References

1	Zysman-Colman Z, Tremblay GM, Bandeali S and Landry JS: Bronchopulmonary dysplasia-trends over three decades. Paediatr Child Health. 18:86–90. 2013.PubMed/NCBI View Article : Google Scholar
2	Jobe AH: What is BPD in 2012 and what will BPD become? Early Hum Dev. 88:S27–S28. 2012.PubMed/NCBI View Article : Google Scholar
3	Thébaud B and Abman SH: Bronchopulmonary dysplasia: Where have all the vessels gone? Roles of angiogenic growth factors in chronic lung disease. Am J Respir Crit Care Med. 175:978–985. 2007.PubMed/NCBI View Article : Google Scholar
4	De Paepe ME, Greco D and Mao Q: Angiogenesis-related gene expression profiling in ventilated preterm human lungs. Exp Lung Res. 36:399–410. 2010.PubMed/NCBI View Article : Google Scholar
5	Stenmark KR and Abman SH: Lung vascular development: Implications for the pathogenesis of bronchopulmonary dysplasia. Annu Rev Physiol. 67:623–661. 2005.PubMed/NCBI View Article : Google Scholar
6	Hartling L, Liang Y and Lacaze-Masmonteil T: Chorioamnionitis as a risk factor for bronchopulmonary dysplasia: A systematic review and meta-analysis. Arch Dis Child Fetal Neonatal Ed. 97:F8–F17. 2012.PubMed/NCBI View Article : Google Scholar
7	Cao L, Wang J, Tseu I, Luo D and Post M: Maternal exposure to endotoxin delays alveolarization during postnatal rat lung development. Am J Physiol Lung Cell Mol Physiol. 296:L726–L737. 2009.PubMed/NCBI View Article : Google Scholar
8	Kim DH, Choi CW, Kim EK, Kim HS, Kim BI, Choi JH, Lee MJ and Yang EG: Association of increased pulmonary interleukin-6 with the priming effect of intra-amniotic lipopolysaccharide on hyperoxic lung injury in a rat model of bronchopulmonary dysplasia. Neonatology. 98:23–32. 2010.PubMed/NCBI View Article : Google Scholar
9	Kume T: Ligand-Dependent notch signaling in vascular formation. Adv Exp Med Biol. 727:210–222. 2012.PubMed/NCBI View Article : Google Scholar
10	Hori K, Sen A and Artavanis-Tsakonas S: Notch signaling at a glance. J Cell Sci. 126:2135–2140. 2013.PubMed/NCBI View Article : Google Scholar
11	Caolo V, Molin DG and Post MJ: Notch regulation of hematopoiesis, endothelial precursor cells, and blood vessel formation: Orchestrating the vasculature. Stem Cells Int. 2012(805602)2012.PubMed/NCBI View Article : Google Scholar
12	Zhan CY, Yuan TM, Sun Y and Yu HM: Early gestational intrauterine infection induces postnatal lung inflammation and arrests lung development in a rat model. J Matern Fetal Neonatal Med. 24:213–222. 2011.PubMed/NCBI View Article : Google Scholar
13	Pan J, Zhan C, Yuan T, Wang W, Shen Y, Sun Y, Wu T, Gu W, Chen L and Yu H: Effects and molecular mechanisms of intrauterine infection/inflammation on lung development. Respir Res. 19(93)2018.PubMed/NCBI View Article : Google Scholar
14	Uenishi GI, Jung HS, Kumar A, Park MA, Hadland BK, McLeod E, Raymond M, Moskvin O, Zimmerman CE, Theisen DJ, et al: NOTCH signaling specifies arterial-type definitive hemogenic endotheliumfrom human pluripotent stem cells. Nat Commun. 9(1828)2018.PubMed/NCBI View Article : Google Scholar
15	Zhao H, Xu CN, Huang C, Jiang J and Li L: Notch1 signaling participates in the release of inflammatory mediators in mouse RAW264.7 cells via activating NF-κB pathway. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 33:1310–1315. 2017.PubMed/NCBI(In Chinese).
16	Wang Y, Chen H, Li H, Zhang J and Gao Y: Effect of angiopoietin-like protein 4 on rat pulmonary microvascular endothelial cells exposed to LPS. Int J Mol Med. 32:568–576. 2013.PubMed/NCBI View Article : Google Scholar
17	Zhou HS, Li M, Sui BD, Wei L, Hou R, Chen WS, Li Q, Bi SH, Zhang JZ and Yi DH: Lipopolysaccharide impairs permeability of pulmonary microvascular endothelial cells via connexin40. Microvasc Res. 115:58–67. 2018.PubMed/NCBI View Article : Google Scholar
18	Zhang K, Wang P, Huang S, Wang X, Li T, Jin Y, Hehir M and Xu C: Different mechanism of LPS-induced calcium increase in human lung epithelial cell and microvascular endothelial cell: A cell culture study in a model for ARDS. Mol Biol Rep. 41:4253–4259. 2014.PubMed/NCBI View Article : Google Scholar
19	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
20	Landry JS, Chan T, Lands L and Menzies D: Long-Term impact of bronchopulmonary dysplasia on pulmonary function. Can Respir J. 18:265–270. 2011.PubMed/NCBI View Article : Google Scholar
21	Krebs LT, Xue Y, Norton CR, Shutter JR, Maguire M, Sundberg JP, Gallahan D, Closson V, Kitajewski J, Callahan R, et al: Notch signaling is essential for vascular morphogenesis in mice. Genes Dev. 14:1343–1352. 2000.PubMed/NCBI
22	Uyttendaele H, Ho J, Rossant J and Kitajewski J: Vascular patterning defects associated with expression of activated notch4 in embryonic endothelium. Proc Natl Acad Sci USA. 98:5643–5648. 2001.PubMed/NCBI View Article : Google Scholar
23	Gale NW, Dominguez MG, Noguera I, Pan L, Hughes V, Valenzuela DM, Murphy AJ, Adams NC, Lin HC, Holash J, et al: Haploinsufficiency of delta-like 4 ligand results in embryonic lethality due to major defects in arterial and vascular development. Proc Natl Acad Sci USA. 101:15949–15954. 2004.PubMed/NCBI View Article : Google Scholar
24	Trindade A, Kumar SR, Scehnet JS, Lopes-da-Costa L, Becker J, Jiang W, Liu R, Gill PS and Duarte A: Overexpression of delta-like 4 induces arterialization and attenuates vessel formation in developing mouse embryos. Blood. 112:1720–1729. 2008.PubMed/NCBI View Article : Google Scholar
25	Qiao LN, Xu HB, Shi K, Zhou TF, Hua YM and Liu HM: Role of notch signal in angiotensin II induced pulmonary vascular remodeling. Transl Pediatr. 2:5–13. 2013.PubMed/NCBI View Article : Google Scholar
26	Muratore CS, Luks FI, Zhou Y, Harty M, Reichner J and Tracy TF: Endotoxin alters early fetal lung morphogenesis. J Surg Res. 155:225–230. 2009.PubMed/NCBI View Article : Google Scholar
27	Kallapur SG, Bachurski CJ, Le Cras TD, Joshi SN, Ikegami M and Jobe AH: Vascular changes after intra-amniotic endotoxin in preterm lamb lungs. Am J Physiol Lung Cell Mol Physiol. 287:L1178–L1185. 2004.PubMed/NCBI View Article : Google Scholar
28	Bhatt AJ, Pryhuber GS, Huyck H, Watkins RH, Metlay LA and Maniscalco WM: Disrupted pulmonary vasculature and decreased vascular endothelial growth factor, Flt-1, and TIE-2 in human infants dying with bronchopulmonary dysplasia. Am J Respir Crit Care Med. 164:1971–1980. 2001.PubMed/NCBI View Article : Google Scholar
29	Hou Y, Liu M, Husted C, Chen C, Thiagarajan K, Johns JL, Rao SP and Alvira CM: Activation of the nuclear factor-κB pathway during postnatal lung inflammation preserves alveolarization by suppressing macrophage inflammatory protein-2. Am J Physiol Lung Cell Mol Physiol. 309:L593–L604. 2015.PubMed/NCBI View Article : Google Scholar
30	Alvira CM: Nuclear factor-kappa-B signaling in lung development and disease: One pathway, numerousfunctions. Birth Defects Res A Clin Mol Teratol. 100:202–216. 2014.PubMed/NCBI View Article : Google Scholar