Anti-inflammatory Effect of 25-OH Vitamin D in Neonatal Infectious Pneumonia by Modulating the TGF-β-mediated YAP/TAZ Nuclear Translocation
10.3969/j.issn.1671-7414.2024.04.006
- VernacularTitle:25-OH维生素D通过调控TGF-β介导的YAP/TAZ核转位机制对新生儿感染性肺炎的抗炎作用研究
- Author:
Yang LIU
1
;
Yun SHI
;
Linping ZHONG
;
Gaoyang QIN
Author Information
1. 电子科技大学医学院附属妇女儿童医院/成都市妇女儿童中心医院新生儿科,成都 611731
- Keywords:
neonatal infectious pneumonia;
25-OH-VD;
TGF-β;
YAP/TAZ nuclear translocation
- From:
Journal of Modern Laboratory Medicine
2024;39(4):28-33,39
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the anti-inflammatory effect of 25-OH vitamin D(25-OH-VD)on neonatal infectious pneumonia(NIP)and its mechanism.Methods A total of 65 children with NIP admitted to Chengdu Women and Children's Central Hospital from January 2022 to January 2023 were selected.According to the severity of the disease,they were divided into mild group(n=34)and severe group(n=31),and 60 healthy neonates in the same period were selected as the control group.Serum 25-OH-VD,interleukin-2(IL-2),interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α)and IL-1β levels were measured by ELISA.The expressions of vitamin D receptor(VDR),transforming growth factor-β(TGF-β)/nuolea Yes-associated protein(n-YAP)and nuclear transcriptional coactivator PDZ-binding motif(n-TAZ)pathway related proteins in peripheral blood mononuclear cells were detected by Western blot.NIP in vitro models were established by lipopolysaccharide(LPS)stimulation of human lung epithelial cells,which were divided into control group,LPS group and LPS+VD group.Cell viability was detected by CCK-8 assay.The mRNA expressions of recombinant cytochrome P450 27B1(CYP27B1)and VDR in each group were detected by qRT-PCR.The number of YAP positive nuclei in each group was detected by immunocytochemistry.The nuclear translocation of YAP/TAZ complex in each group was detected by immunofluorescence.The expression of inflammatory factor-related proteins in each group was detected by Western blot.Results Compared with the control group,the serum levels of IL-2(1.91±0.18 μ g/L,2.63±0.27 μg/L vs 1.05±0.12 μg/L),IFN-γ(1.73±0.13 μg/L,2.18±0.19 μg/L vs 1.03±0.07 μg/L),TNF-α(1.79±0.08 μg/L,2.38±0.13 μg/L vs 0.97±0.04 μg/L),IL-1 β(2.18±0.07 μg/L,2.59±0.11μg/L vs 0.96±0.02 μg/L),TGF-β(1.67±0.21,2.43±0.42 vs 1.02±0.04),n-YAP(2.08±0.11,4.23±0.37 vs 0.99±0.02)and n-TAZ(2.47±0.42,4.21±0.58 vs 1.03±0.05)proteins of the children in the mild and severe groups were gradually increased,but the content of 25-OH-VD(12.57±2.21 μg/L,7.85±2.03 μg/L vs 16.76±1.02 μg/L)and the expression of VDR(0.73±0.09,0.51±0.06 vs 1.03±0.08)proteins were gradually decreased,and the differences were significant(F=18.983～56.782,all P＜0.001).Cell experiment results showed that cellular survival rate was lower at 12 h(76.23％±0.73％vs 116.72％±2.14％),24 h(57.23％±0.94％vs 125.76％±1.67％)and 48 h(41.23％±0.56％vs 138.56％±1.35％)in the LPS group compared with the control group(t=10.342,26.562,37.821),but the rate of cytosolic n-YAP positivity(47.35±3.47 vs 12.46±1.34),the rate of nuclear translocation of the YAP/TAZ complex(2.56％±0.32％vs 1.01％±0.06％)(t=46.362,26.921),the protein expression levels of IL-2(2.03±0.09vs 1.03±0.08),IFN-γ(2.07±0.21 vs 1.02±0.04),TNF-α(2.18±0.11 vs 0.99±0.02)and IL-1β(3.17±0.42 vs 1.03±0.05)were up-regulated(t=28.341,26.713,31.235,47.823),with significant differences(all P＜0.001),while there was no significant difference in the mRNA expression of CYP27B1 and VDR(t=0.872,0.786,all P＞0.05).Compared with the LPS group,the cell survival rate at 12h(85.23％±0.36％),24h(79.82％±0.63％)and 48h(76.28％±0.72％)and the mRNA expression of CYP27B1(4.42±0.14)and VDR(5.13±0.56)were elevated in the LPS+VD group,while the nucleus n-YAP positivity(24.41±3.23),nuclear translocation of the YAP/TAZ complex(1.47％±0.26％),IL-2(1.21±0.06),IFN-γ(1.13±0.42),TNF-α(1.03±0.37)and IL-1β(1.61±0.58)protein levels were down-regulated,and the differences were significant(t=7.263,19.892,23.145,27.872,26.982,14.762,13.623,18.273,25.314,27.873,22.134,all P＜0.0 1).Conclusion The serum 25-OH-VD level is related to the severity of NIP.Exogenous VD supplementation may play an anti-inflammatory role in reducing NIP injury by regulating the TGF-β-mediated YAP/TAZ nuclear translocation mechanism.
