Table 1.

Effects of HNE and 15d-PGJ₂ on endothelial cells.

Cell type	Treatment	Conc.⁎ (μmol/L)	Time	Main findings	References
RTK and MAPK activation
HEC	HNE	0.1	3 h	HNE (0.1 µM) and oxLDL (200 µg/ml)↑EGFR activation	[36]
	oxLDL				[49]
BLMVEC	HNE	10–100	2 h	↑Phosphorylation of ERK, JNK, p38 MAPK
MPEC	HNE	20	0–2 h	↑p∼JNK within 30 min,↔p∼p38, p∼ERK1/2	[20]
BLMVEC	HNE	25	30 min	↑MAPK activation	[88]
BPAEC	HNE	50	30 min	↑Phospholipase D activation via protein tyrosine phosphorylation	[37,141]
HUVEC	15d-PGJ2	10	2 h	↑Phosphorylation of MAPK JNK (2 h)	[142]
Transcriptional activity
HUVEC	HNE	5	12 h	↑HO-1 and NQO1 via Nrf2, protects HUVEC from tBHP attack	[55]
HUVEC	HNE	1–10	12 h	↔NFκB activation, ↓IL-8 and ICAM-1 production in a concentration dependent manner indicating ↓inflammatory responses	[10]
HUVEC	15d-PGJ2	2.5	2–8 h	↑Nrf2 nuclear accumulation, ↑HO-1, ↑NQO1, ↑adducts with Keap1 at cysteines residues in IVR region, which may mediate shear induced Nrf2 activation	[143]
BAEC	15d-PGJ2	2	16 h	↑GSH, HO-1, ↑Keap1 adduct formation	[144]
HUVEC	15d-PGJ2	5	24 h	↑GSH, ↑GCLC, ↑GCLM, ↑resistance to oxidative stress which relies on de novo GSH synthesis, ARE responsible for GCLC induction, Cys273 and Cys278 of Keap1 conjugated by 15d-PGJ2	[52,145]
Oxidative stress and protein adduct clearance
BAEC	HNE	5	1 h	Modifies thioredoxin-1 at cys-73 and ↓activity, ↑ROS, ↓GSH	[90]
HAEC	HNE	5	24 h	Protects HAEC from 6-hydroxydopamine induced cell death
BAEC	HNE	25	4 h	Proposed ↑proteasomal degradation of GTPCH and Hsp90	[23]
Cellular dysfunction
BLMVEC	HNE	10–100	2 h	↑Endothelial permeability; ↑Michael adducts formation, actin fiber remodeling via phosphorylation of ERK, JNK, p38 MAPK	[49]
BLMVEC	HNE	25	30 min	Induces actin rearrangement, ↓GSH, ↑cell adhesion and surface integrins; ↓tyrosine phosphorylation of FAK,↑MAPK activation, ↑Michael adducts with ↑focal adhesion & adherens junctional proteins, reversed by pretreatment NAC	[88]
BAEC	HNE	5	1 h	↑Monocyte adhesion	[90]
BAEC	HNE	10	5 d	↑Membrane phospholipid perturbation, ↑prostacyclin, ↑monocyte migration	[146]
BAEC	HNE	10	6–8 h	↓Junctional communication	[87]
BAEC	HNE	25	4 h	↑ROS, ↓NO, GSH, ↑apoptosis involving caspase-3 activation which can be attenuated by BH4 supplementation. ↓BH4 by ↓GTPCH leading to ↑eNOS uncoupling, ↓HSP90 leading to ↓eNOS phosphorylation, proposed ↑proteasomal activity and proteasomal degradation of BH4 and HSP 90	[23]
Mitochondrial function
BAEC	HNE	5–10	4–16 h	↑GSH and HO-1 at 16 h, ↑ROS from mitochondria after 4 h	[71]
HUVEC	15d-PGJ2	2.5	24 h	↑GSH, ↑complex I activity in cell extracts, dependent on de novo protein synthesis in response to ROS but independent of GSH depletion	[94]
BAEC	15d-PGJ2	10	16 h	↑HO-1, ↓mitochondrial membrane potential ↑Nrf2 nuclear accumulation, blocked IBTP, a mitochondrial-targeted thiol reactive compound.	[53]
ER stress
HUVEC	HNE	25	30 min – 2 h	↑Protein adduction (HSP90, HSP70,PDI), ↑xBP-1 splicing, ↑PERK, p∼eIF2α, and ATF6 translocation; ↑Grp78 and HERP, ↑ICAM-1, cell adhesion, TNF-α, IL-6, and IL-8; activation of endothelial cells mediated by ↑ER stress, noting depletion of GSH cannot ↑ER stress	[107]
HMEC	HNE	20	24 h	↑PERK and ATF6; ↑p∼IRE1 and p∼eIF2α; ↑ER stress, which is prevented by pretreatment with NAC	[108]
HMEC	HNE/oxLDL	0–25 (HNE)/ 200 µg/ml oxLDL	14–18 h	Forms adducts with PDI,↓PDI activity and↓cell viability. ↑CHOP and xBP1s mRNA as indicators of ↑ER stress. Loss of PDI activity and reduced viability prevented by NAC.	[110]
Apoptosis
HUVEC	HNE	10–50	3 h	↑Cell death and ↓growth capacity but this effect is affected by serum in the medium	[147]
HUVEC	HNE	10	12 h	↑Apoptosis, ↓protein synthesis	[10]
PCEC	HNE	1–50	3 h	↑Chromosomal aberrations and micronuclei formation	[113]
BAEC	HNE	25	4 h	↑ROS, ↓NO,↓GSH, ↑apoptosis involving caspase-3 activation which can be attenuated by BH4 supplementation.	[23]
				↓BH4 resulting from ↓GTPCH and ↓HSP90 levels leads to ↑eNOS uncoupling and ↓eNOS phosphorylation. Proposed ↑proteasomal activity towards GTPCH and HSP 90 underlies impaired NO production
MPEC	HNE	40	8 h	↑p∼JNK leading to ↑p53 and Bax expression and↑apoptosis. Abolished by GST4 overexpression	[20]
HUVEC	15d-PGJ2	10	2–16 h	↑ROS (1 h), ↑phosphorylation of MAPK and JNK (2 h), ↑p53 expression and phosphorylation (8 h), ↑caspase associated apoptosis	[142]

Cell type abbreviations: MPEC, mouse pancreatic islet endothelial cells; HEC human endothelial cell line CRL-1998; BAEC, bovine aortic endothelial cells; BLMVEC, Bovine lung microvascular vein endothelial cells; BPAEC, bovine pulmonary arterial endothelial cells; HMEC, human microvascular endothelial cells; HUVEC, human umbilical vein endothelial cells; PCEC, porcine cerebral endothelial cells.