TABLE 2.

In vitro and in vivo studies outcome.

1	García et al. (2017)	To study effects of Hydroxytyrosol in endothelial cell expressing extracellular matrix remodeling enzymes in inhibition of angiogenesis	Animal and Cells 1. Rats 2. BAECs Treatment In vitro-HT 0–800 nmol) and 1 mM of HT for 24 h cells In vivo-HT 31.2,62.5, 125 and 250 µm) for 48 hours	1. Ex vivo rat aortic ring assay 2. In vivo chorioallantoic membrane (CAM) assay 3. mRNAs for some extracellular matrix remodeling enzymes	1. HT reduced MMP-1 and MMP-2, uPA mRNA expression 2. HT inhibit ex vivo angiogenesis, yet endothelial outgrowing observed 3. HT prevented in vivo angiogenesis		HT decreased expression of extracellular matrix remodeling enzyme that could supress migration of smooth muscle cells
2	Catalán et al. (2018)	To study the potential of hydroxytyrosol (HT) and its plasmatic metabolites (HTmet) in enhancement of endothelial function	Animal and cells 1. Apolipoprotein E knockout mice 2. HAEC 3. Jurkat Treatment Invivo-10 mg/kg/day of HT derivatives for 12 weeks Invitro-cells (1, 2 and 5 µM) and TNF-α (10 ng/ml) for 24 h	1. VCAM-1, E-selectin, MCP-1, ICAM-1 expression 2. Human Phospho-MAPK Array 3. NF-B (p65) expression	1. Mice aortas stained less for E-selectin, MCP-1, and ICAM-1 2. HTmet reduced Jurkat T adhesion 3. HTmet decreased E-selectin and VCAM-1 mRNA expression in HAECs 4. HT and HTmet decreased CREB, ERK, JNK pan, JNK, p38δ, p70 S6 kinase	1. ERK 2. JNK 3. MAPK	HT and its metabolites shown to have endothelial protection potential which regulated by the MAPK pathway
3	Yaoa et al. (2019)	To examine the potential of hydroxytyrosol acetate on vascular endothelial inflammation mechanism	Animal and Cells 1. Specific Sirt6 knockout mice hypercholesteraemic 2. HUVECs Treatment Invivo- P-407 (0.5 g/kg), P-407 + HT (5, 10, 20 mg/kg), and P-407+HT-AC (5, 10, 20 mg/kg) groups Invitro-HT or HT-AC (25, 50, or 100 μmol/L) for 1 h, and then stimulated with TNF (10 ng/ml) for 8 h	1. Cell viability 2. SOD, MDA and ROS level 3. SIRT6 siRNA transfection 4. SIRT6 and PKM2 expression 5. HT-AC molecular docking	1. HT and HT-AC decreased TNF and IL1B in mice serum 2. HT and HT-AC decreased mRNA expression of Il-b, Il6 and Ccl2 and TNF 3. HT and HT-AC decreased mRNA expressions of IL1B, IL6 and CCL2 in HUVECs 4. HT-AC increased SOD while decreased MDA and ROS level in TNF- induced HUVECs 5. HT-AC decreased TNFRSF1A protein and mRNA in HUVECs 6. HT-AC upregulated SIRT6 protein and mRNA expression in mice 7. Molecular docking shown good compatibility between HT-AC and SIRT6 8. HT-AC decreased expression of PKM2 in mice and TNF-stimulate HUVECs	1. PKM2	HT and HT-AC exhibited protection against endothelial inflammation in mice and HUVECs cells by mediating PKM2 signaling pathway
4	Fuccelli et al. (2018)	To study the effect of HT in inflammatory markers Cyclooxygenase-2 (COX2) And tumor necrosis factor alfa (TNF-α) and oxidative stress reduction in vivo systematic inflammation model	Animal Balb/c mice Treatment 1. HT (40 and 80 mg/kg) 2. LPS induction (50 µg/mouse)	1. COX2 mRNA detection 2. TNF-a cytokine determination 3. DNA damage assessment 4. Antioxidant plasma power quantification	1. HT inhibits the COX2 gene expression 2. HT reduces the TNF-α cytokine secretion 3. HT improves the antioxidant power of plasma 4. HT prevents the DNA damage induced	1. COX2 2. TNF-α	HT inhbited LPS induced COX2 expression, TNF-α production and the DNA damage while enhance antioxidant potential of plasma in vivo model

Abbreviations: THP-1, human monocyte cell line; U937, Monocytic cell line; HUVECs, Human umbilical vein endothelial cells; HMEC-1, Human microvascular endothelial cell line; PBMC, Human peripheral blood mononuclear cells; PPAECs, Porcine pulmonary artery endothelial cells; BVSMC, Bovine Vascular smooth muscle cells; HMECs, Human microvascular endothelial cells; VAFs, vascular adventitial fibroblasts; HVECs, Human vascular endothelium cells; BAECs, Bovine aorta endothelial cells; HAECs, human aortic endothelial cells; EA, hy926-endothelial cells; C2C12, myoblasts cells; HREC, Human retinal endothelial cells; RVSMCs, Rat Vascular smooth muscle cells; PMA, phorbol myristate acetate; MMP, matrix metalloproteinase; ROS, Reactive oxygen species; COX-2, cyclooxygenase 2; NF-κβ, nuclear factor kappa-light-chain-enhancer of activated B cells; MCP-1, monocyte chemoattractant protein-1; ICAM-1, intercellular adhesion molecule-1; VCAM-1, vascular cell adhesion molecule-1; IL-1β, interleukin-1β; TNF-α, tumour necrosis factor-α; HMVECs-d-Ad, Human dermal microvascular endothelial cells; VEGF, Vascular endothelial growth factor; prostaglandin (PG)E2; protein kinase C (PKC); FOXO3a, forkhead transcription factor 3a; AMPK-AMP, activated protein kinase; Akt, protein kinase B; CORM-2, Carbon Monoxide-Releasing Molecule-2; PEMF, Pulsed electromagnetic fields; mTOR-mechanistic target of rapamycin; TGF-β1, Transforming growth factor; MAPK, mitogen-activated protein kinase; EndMT, Endothelial-to-mesenchymal transition; HT-3Os, plasma metabolite HT-3O sulfate; FGFR1, fibroblast growth factor receptor 1