Table 4.

Functional role for EVs in APAP toxicity.

Model/EV source	Dose (APAP)	Isolation method	EV marker	Result	References [#]
PRH Cell media	10 mmol/L Collected 36 h later	100,000 g UC	None reported	Metabolically active APAP-sEVs produce unique metabolites in serum	[126]
C57BL/6J Plasma	300 mg/kg Collected plasma 2, 4, 6 h PT	100,000 g UC	ALIX/CD63	Increased EVs at 2 h prior to change in ALT. No APAP-CYS adducts present in EV fraction. Increased plasma-EVs in APAP overdose patients	[127]
PRH Cell media	10 mmol/L Collected 36 h later	Size-exclusion chromatography	CD63	Cyp2d1 is present in EVs and metabolically active	[131]
PRH Cell media	10 mmol/L Collected 36 h later	100,000 g UC	Hsp70/Hsp90/Rab8	Increased Arginine synthase, Arginase 1, GST, cysteine synthesis	[131]
C57BL/6 Ja Plasma	300 mg/kg Collected from plasma 24 h later	Exoquick (commercial)	CD63/TSG101	In vitro: Increased cell death in vivo: Increased TNF-α, IL-1β, JNK activation, no change in ALT	[129]
BALB/c Plasma	300 mg/kg Collected from plasma 1 and 3 h post treatment	Modified Exoquick	Hsp90/Hsp70/CD63	GST packaged in plasma APAP-EVs	[133]
PRH Cell media	10 mmol/L Collected 36 h later	100,000 g UC, SG 1.18–1.23 g/mL	ALIX/CD81/CD63/Hsp70/RAB8	Hepatic-EV can increase deoxyinosine, APAP-EVs can deplete arginine and interfere with endothelial function	[134]
Human Plasma	–	Plasma Exo Kit (commerical)	CD63	Cyp2e1 is metabolically active and packaged into plasma EVs	[128]
PRH Cell media	–	100,000 g UC	CD81/TSG101/Flotillin 1/AIP1	Defined three subpopulations of EVs from PRH. Evidence EVs can transfer mRNA to stellate-like cells and activate them	[12]
Human Plasma	–	Exoquick (commercial)	CD63/CD81	Human plasma contains functional Cyp2e1	[130]
Winstar Rats Urine	–	100,000 g UC, SG	CD63/CD81/Flotillin/TSG101 Albumin negative	GGT present in urinary vesicles	[135]

^aStudy treated mice with 300 mg/kg APAP, collected EVs from plasma, then took these APAP-EVs and treated to primary mouse hepatocytes or i.p. injection to mice.