Inflammatory infiltration in aortic valves promotes calcific aortic valve disease (CAVD) progression. While soluble extracellular matrix (ECM) proteins induce inflammatory responses in aortic valve interstitial cells (AVICs), the impact of monocytes on AVIC inflammatory responses is unknown. We tested the hypothesis that monocytes enhance AVIC inflammatory responses to soluble ECM protein in this study.
Methods
Human AVICs isolated from normal aortic valves were cocultured with monocytes and stimulated with soluble ECM protein (matrilin-2). ICAM-1 and IL-6 productions were assessed. YAP and NF-κB phosphorylation were analyzed. Recombinant CD18, neutralizing antibodies against β2-integrin or ICAM-1, and inhibitor of YAP or NF-κB were applied.
Results
AVIC expression of ICAM-1 and IL-6 was markedly enhanced by the presence of monocytes, although matrilin-2 did not affect monocyte production of ICAM-1 or IL-6. Matrilin-2 up-regulated the expression of monocyte β2-integrin and AVIC ICAM-1, leading to monocyte-AVIC adhesion. Neutralizing β2-integrin or ICAM-1 in coculture suppressed monocyte adhesion to AVICs and the expression of ICAM-1 and IL-6. Recombinant CD18 enhanced the matrilin-2-induced ICAM-1 and IL-6 expression in AVIC monoculture. Further, stimulation of coculture with matrilin-2 induced greater YAP and NF-κB phosphorylation. Inhibiting either YAP or NF-κB markedly suppressed the inflammatory response to matrilin-2 in coculture.
Conclusion
Monocyte β2-integrin interacts with AVIC ICAM-1 to augment AVIC inflammatory responses to soluble matrilin-2 through enhancing the activation of YAP and NF-κB signaling pathways. Infiltrated monocytes may promote valvular inflammation through cell–cell interaction with AVICs to enhance their sensitivity to damage-associated molecular patterns.
A novel magnetic (Fe3O4) surface molecularly imprinted polymer (MIP) based on ionic liquid (IL) (Fe3O4@VTEO@IL-MIPs) was prepared for the selective extraction of lysozyme (Lys). As the functional monomer of the MIPs, an imidazolium-based IL with vinyl groups was prepared. It can provide multiple interactions with template molecules. The amount of IL was optimized (200 mg). Fourier transform infrared spectrometry (FT-IR), transmission electron microscopy (TEM), dynamic light scattering (DLS), thermogravimetric analysis (TGA) and a vibrating sample magnetometer (VSM) were used to characterize the MIP. The results indicate the successful formation of an imprinting polymer layer. The concentration of Lys in the supernatant was determined by UV-vis spectrophotometry at a wavelength of 280 nm. The maximum adsorption capability of the MIP is 213.7 mg g−1 and the imprinting factor (IF) is 2.02. It took 2.5 h for the MIP to attain adsorption equilibrium. The structure of the protein was evaluated using circular dichroism (CD) spectra and UV-visible spectra. The adsorption performance was further investigated in detail by selective adsorption experiments, competitive rebinding tests, and reusability and stability experiments. Furthermore, it was utilized to separate the template protein from a mixture of proteins and real samples successfully because of the high adsorption capacity for Lys.A novel magnetic (Fe3O4) surface molecularly imprinted polymer (MIP) based on ionic liquid (IL) (Fe3O4@VTEO@IL-MIPs) was prepared for the selective extraction of lysozyme (Lys).相似文献