In:
PLOS ONE, Public Library of Science (PLoS), Vol. 16, No. 8 ( 2021-8-3), p. e0255656-
Abstract:
Although quiescent hepatic stellate cells (HSCs) have been suggested to regulate hepatic blood flow, there is no direct evidence that quiescent HSCs display contractile abilities. Here, we developed a new method to quantitatively measure the contraction of single isolated HSCs and evaluated whether endothelin-1 (ET-1) induced contraction of HSCs in a non-activated state. HSCs isolated from mice were seeded on collagen gel containing fluorescent beads. The beads around a single HSC were observed gravitating toward the cell upon contraction. By recording the movement of each bead by fluorescent microscopy, the real-time contraction of HSCs was quantitatively evaluated. ET-1 induced a slow contraction of non-activated HSCs, which was inhibited by the non-muscle myosin II inhibitor blebbistatin, the calmodulin inhibitor W-7, and the ET A receptor antagonist ambrisentan. ET-1-induced contraction was also largely reduced in Ca 2+ -free conditions, but sustained contraction still remained. The tonic contraction was further diminished by the Rho-kinase inhibitor H-1152. The mRNA expression of P/Q-type voltage-dependent Ca 2+ channels (VDCC), as well as STIM and Orai , constituents of store-operated channels (SOCs), was observed in mouse non-activated HSCs. ET-1-induced contraction was not affected by amlodipine, a VDCC blocker, whereas it was partly reduced by Gd 3+ and amiloride, non-selective cation channel blockers. However, neither YM-58483 nor SKF-96365, which inhibit SOCs, had any effects on the contraction. These results suggest that ET-1 leads to Ca 2+ -influx through cation channels other than SOCs and produces myosin II-mediated contraction of non-activated HSCs via ET A receptors, as well as via mechanisms involving Ca 2+ -calmodulin and Rho kinase.
Type of Medium:
Online Resource
ISSN:
1932-6203
DOI:
10.1371/journal.pone.0255656
DOI:
10.1371/journal.pone.0255656.g001
DOI:
10.1371/journal.pone.0255656.g002
DOI:
10.1371/journal.pone.0255656.g003
DOI:
10.1371/journal.pone.0255656.g004
DOI:
10.1371/journal.pone.0255656.g005
DOI:
10.1371/journal.pone.0255656.t001
DOI:
10.1371/journal.pone.0255656.s001
DOI:
10.1371/journal.pone.0255656.s002
DOI:
10.1371/journal.pone.0255656.s003
DOI:
10.1371/journal.pone.0255656.s004
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2021
detail.hit.zdb_id:
2267670-3
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