4-hexylresorcinol-induced protein expression changes in human umbilical cord vein endothelial cells as determined by immunoprecipitation high-performance liquid chromatography

4-Hexylresorcinol (4HR) is used as a food preservative and an ingredient of toothpaste and cosmetics. The present study was performed using 233 antisera to determine the changes in protein expression induced by 4HR in human umbilical cord vein endothelial cells (HUVECs), and evaluated the 4HR-induced effects in comparison with previous results (Kim et al., 2019). Similar to RAW 264.7 cells, 4HR-treated HUVECs showed decreases in the expression of the proliferation-related proteins, cMyc/MAX/MAD network proteins, p53/RB and Wnt/β-catenin signaling, and they showed inactivation of DNA transcription and protein translation compared to the untreated controls. 4HR upregulated growth factors (TGF-β1, β2, β3, SMAD2/3, SMAD4, HGF-α, Met, IGF-1) and RAS signaling proteins (RAF-B, p38, p-p38, p-ERK-1, and Rab-1), and induced stronger expression of the cellular protection-, survival-, and differentiation-related proteins in HUVECs than in RAW 264.7 cells. 4HR suppressed NFkB signaling in a manner that suggests potential anti-inflammatory and wound healing effects by reducing M1 macrophage polarization and increasing M2 macrophage polarization in both cells. 4HR-treated HUVECs tended to increase the ER stress mediators by upregulating eIF2AK3, ATF4, ATF6, lysozyme, and LC3 and downregulating eIF2α and GADD153 (CHOP), resulting in PARP-1/AIF-mediated apoptosis. These results indicate that 4HR has similar effects on the protein expression of HUVECs and RAW 264.7 cells, but their protein expression levels differ according to cell types. The 4HR-treated cells showed global protein expression characteristic of anticancer and wound healing effects, which could be alleviated simultaneously by other proteins exerting opposite functions. These results suggest that although 4HR has similar effects on the global protein expression of HUVECs and RAW 264.7 cells, the 4HR-induced molecular interferences in those cells are complex enough to produce variable protein expression, leading different cell functions. Moreover, HUVECs have stronger wound healing potential to overcome the impact induced by 4HR than RAW 264.7 cells.

Effect of anthocyanin-rich sour cherry extract on the level of IL-8 in LPS-induced endothelial cell

The anthocyanin content of the Hungarian sour cherry is remarkable. Nutraceutical and pharmaceutical effects of the anthocyanins and their role in disease prevention have been studied extensively. Endothelial cells are involved in the pathogenesis of several inflammatory diseases. The objective of this work was to investigate pure sour cherry extract on human umbilical cord vein endothelial cells (HUVECs) as an inflammatory model. HUVECs were treated with 100 ng/mL lipopolysaccharide (LPS) and 50 mg/mL sour cherry extract or M199 medium as control. The optimal concentration range of the sour cherry extract was investigated and selected based on MTT assay measuring the conversion of the tetrazolium salt to formazan by mitochondrial dehydrogenases. The level of interleukine-8 (IL-8), a pro-inflammatory cytokine, was measured in Luminex MagPlex assay. LPS treatment significantly increased the secretion of IL-8. The pure sour cherry extract was able to attenuate this increment indicating the potent anti-inflammatory effect of pure sour cherry extract. Our results emphasize that pure sour cherry extract could reduce the LPS-induced inflammatory response thereby may improve endothelial dysfunction.

Determining the cytotoxic effect potential of ozonated hazelnut oil

Although the use of hazelnut oil is becoming ever more common, the ozonated form of this oil has never been obtained before. Ozonated vegetable oils are generally preferred in ozone therapy as they are easy to apply and obtain. In this study, the density, viscosity, peroxide and iodine values of hazelnut oil obtained by being treated with ozone in different periods at a flow rate of 7-8 were determined and compared with its commercial form, i.e. refined hazelnut oil. In order to determine whether ozonated hazelnut oil can be safely used, its cytotoxic effect potential was assessed by performing 3-[4,5-dimethylthiazol-2-yl]-2,5- diphenyltetrazolium assay on three different cell lines. For this purpose, the viability at five different concentrations of each sample was calculated after 48 and 72 hours of incubation in the H1299 lung cancer, human umbilical cord vein endothelial cells and A549 cell line. As a result, it was found that hazelnut oil applied to three different cell lines and in five different concentrations (100/50/25/12.5/6.25μg/mL) did not have any cytotoxic effects. Furthermore, it was also revealed that the samples did not have a toxic effect due to treatment with ozone in comparison to refined ozonated hazelnut oil. Therefore, this study has shown that ozonated hazelnut oil can be used as an alternative to refined hazelnut oil without exerting any toxic effects due to treatment with ozone. In light of this information, it was concluded that hazelnut oil can be safely used after treatment with ozone without having any cytotoxic effects and that it can be produced as a functional oil providing both the pharmacological and cosmetic effects of ozone.