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CD40 ligand induces expression of vascular cell adhesion molecule 1 and E-selectin in orbital fibroblasts from patients with Graves’ orbitopathy

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Abstract

Purpose

The aim of this study was to detect the effect of the CD40 ligand (CD40L) on the expression of vascular cell adhesion molecule 1 (VCAM-1) and E-Selectin in orbital fibroblasts (OFs) from patients with Graves’ orbitopathy (GO), as well as the signaling pathways involved in this effect.

Methods

OFs were isolated from orbital tissues obtained from patients with severe GO who were undergoing orbital decompression surgery. VCAM-1 and E-selectin RNA and protein expression levels were quantified in OFs stimulated with soluble CD40L (sCD40L). RNA and protein quantification was performed with real-time polymerase chain reaction (PCR) and western blot analysis. Cytoplasmic and nuclear fractions were isolated in order to detect the nuclear translocation of nuclear factor-κB (NF-κB). Signaling pathway inhibitors were applied to determine the pathways involved.

Results

Compared to unstimulated OFs, the mRNA and protein levels of VCAM-1 and E-selectin in OFs incubated with sCD40L were significantly increased. This was observed in dose- and time-course experiments, and the inductive effects of sCD40L were much weaker in OFs from healthy donors. At the same time, we observed that CD40L induced nuclear translocation of NF-κB, also in a dose- and time-dependent manner. The up-regulation of VCAM-1 and E-selectin, as well as the NF-κB nuclear translocation induced by CD40L, was significantly attenuated by inhibitors targeting mitogen-activated protein kinases (MAPKs), phosphatidylinositol 3-kinase (PI3K), and NF-κB.

Conclusions

CD40L demonstrated the ability to up-regulate the expression of VCAM-1 and E-selectin at the pre-translational level in OFs from patients with GO. The MAPK and PI3K pathways and NF-κB may play important roles in CD40L-induced VCAM-1 and E-selectin expression.

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Acknowledgments

Our work is funded by the National Nature and Science Foundation of China (grant Nos. 81070758 and 81371056). Dr. Stephen J. Atkins from the Kellogg Eye Center at the University of Michigan provided English language assistance in the writing of our paper.

Compliance with ethical standards

ᅟ

Conflicts of interest

The authors declare that they have no conflict of interest.

Research involving human participants

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

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Authors and Affiliations

  1. Department of Ophthalmology, Shanghai Changzheng Hospital, Second Military Medical University, 415 Fengyang Road, 200003, Shanghai, China

    Hao Wang, Jin-Wei Cheng, Ji-Ping Cai, You Li, Xiao-Ye Ma & Rui-Li Wei

  2. Department of Transfusion Medicine, Shanghai Changhai Hospital, Second Military Medical University, 174 Changhai Road, Shanghai, China

    Li-Shuang Zhu

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  1. Hao Wang
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Corresponding author

Correspondence to Rui-Li Wei.

Additional information

Hao Wang and Li-Shuang Zhu contributed equally to this work.

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Wang, H., Zhu, LS., Cheng, JW. et al. CD40 ligand induces expression of vascular cell adhesion molecule 1 and E-selectin in orbital fibroblasts from patients with Graves’ orbitopathy. Graefes Arch Clin Exp Ophthalmol 253, 573–582 (2015). https://doi.org/10.1007/s00417-014-2902-1

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  • DOI: https://doi.org/10.1007/s00417-014-2902-1

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