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Microvesicles derived from TGF-β1 stimulated hepatic stellate cells aggravate hepatocellular injury.
Stem Cells Dev. 2019 May 29;:
Authors: Liang Y, Pan Q, Wang R, Ye Z, Li Z, Zeng L, Chen Y, Ma X, Li M, Miao H
Abstract
Hepatic stellate cells (HSCs) are liver specific cells playing critical roles in liver physiological and pathophysiological processes. Transforming growth factor-β1 (TGF-β1) is an inflammatory cytokine secreted by both hepatocytes and HSCs. We have previously shown that microvesicles (MVs) derived from quiescent HSCs protect hepatocyte functions. Here, we investigated the effects of MVs released from TGF-β1 stimulated HSCs (HSC-MVs) on xenobiotic-injured hepatocytes. Two hepatocytes cell lines (BRL-3A and HL-7702) were treated with N-acetyl-p-aminophenol n-(APAP) or H2O2 to build the injury models. Different concentration of HSC-MVs were used to co-culture with injured hepatocytes. MTT, Hochest33258 staining and flow cytometry were used to determine their effects on the viability and apoptosis of hepatocytes. Liver injury indicators, alanine aminotransferase (ALT) and aspartate amino transferase (AST) were assessed by enzyme-linked immune sorbent assay kits. The PI3K activator (740Y-P) and Erk1/2 activator (PDGF-BB) were used for pathway analysis. The expression levels of p-PI3K/PI3K, p-Akt/Akt and activated caspase-3 were measured by Western blot. Results showed that: 1) HSC-MVs dose-dependently impaired the viability of hepatocytes in both injury models; 2) Moreover, HSC-MVs dose-dependently increased the apoptosis in those cell models; 3) HSC-MVs also elevated the levels of ALT and AST in the coculture media; 4) These effects were accompanied by a decrease in p-PI3K/PI3K and p-Akt/Akt, which could be partially abolished by 740Y-P. Meanwhile, the pro-apoptotic effect of HSC-MVs was associated with p-Erk1/2/Erk1/2 down-regulation and activated caspase-3 up-regulation, and could be inhibited by Erk1/2 activation. Our findings demonstrate that HSC-MVs are involved in inflammatory hepatocytes injury probably via the PI3K/Akt, Erk1/2 pathway, and caspase-3.
Hepatic stellate cells (HSCs) are liver specific cells playing critical roles in liver physiological and pathophysiological processes. Transforming growth factor-β1 (TGF-β1) is an inflammatory cytokine secreted by both hepatocytes and HSCs. We have previously shown that microvesicles (MVs) derived from quiescent HSCs protect hepatocyte functions. Here, we investigated the effects of MVs released from TGF-β1 stimulated HSCs (HSC-MVs) on xenobiotic-injured hepatocytes. Two hepatocytes cell lines (BRL-3A and HL-7702) were treated with N-acetyl-p-aminophenol n-(APAP) or H2O2 to build the injury models. Different concentration of HSC-MVs were used to co-culture with injured hepatocytes. MTT, Hochest33258 staining and flow cytometry were used to determine their effects on the viability and apoptosis of hepatocytes. Liver injury indicators, alanine aminotransferase (ALT) and aspartate amino transferase (AST) were assessed by enzyme-linked immune sorbent assay kits. The PI3K activator (740Y-P) and Erk1/2 activator (PDGF-BB) were used for pathway analysis. The expression levels of p-PI3K/PI3K, p-Akt/Akt and activated caspase-3 were measured by Western blot. Results showed that: 1) HSC-MVs dose-dependently impaired the viability of hepatocytes in both injury models; 2) Moreover, HSC-MVs dose-dependently increased the apoptosis in those cell models; 3) HSC-MVs also elevated the levels of ALT and AST in the coculture media; 4) These effects were accompanied by a decrease in p-PI3K/PI3K and p-Akt/Akt, which could be partially abolished by 740Y-P. Meanwhile, the pro-apoptotic effect of HSC-MVs was associated with p-Erk1/2/Erk1/2 down-regulation and activated caspase-3 up-regulation, and could be inhibited by Erk1/2 activation. Our findings demonstrate that HSC-MVs are involved in inflammatory hepatocytes injury probably via the PI3K/Akt, Erk1/2 pathway, and caspase-3.
PMID: 31140359 [PubMed - as supplied by publisher]
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