Isolation and characterization of hepatic mast cells from cholestatic rats.
Isolation and characterization of hepatic mast cells from cholestatic rats.
复制标题
从胆固性大鼠中分离和表征肝肥大细胞。
DOI：
10.1038/labinvest.2016.89
复制
发表时间：
2016-11
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
作者：
Mast cells (MCs) are immune cells that release histamine and other mediators. MC number increases after bile duct ligation (BDL) and blocking mast cell-derived histamine decrease biliary proliferation. We aimed to isolate and characterize MCs from cholestatic livers. Rats were subjected to BDL starting at 6 hrs and up to 14 days. MC infiltration was evaluated by toluidine blue. BDL rats were perfused using standard collagenase perfusion. Following enzymatic digestion, tissue was passed through a fine gauge needle. Suspensions were incubated with MAb AA4, washed and incubated with goat anti-mouse coated Dynal® beads. MCs were stained with toluidine blue, and in isolated MCs, the expression of FCεRI and MC proteases was measured. The expression of histidine decarboxylase, histamine receptors, VEGF-receptors and TIE 1 and 2 was evaluated by qPCR. Histamine and VEGF-A secretion was measured in MC supernatants. MC purity was evaluated by CK-19, CK-8, albumin, VAP-1 and α-SMA expression. In vitro, cholangiocytes and HSCs were treated with isolated MC supernatants from BDL rats treated with either NaCl or cromolyn sodium (to block MC histamine release) and biliary proliferation and hepatic fibrosis were measured. MCs infiltrate the liver and surround bile ducts starting at day 2. We isolated a virtually pure preparation of mature, functional MCs. TEM images reveal distinct secretory granules and isolated MCs secrete histamine. MCs express FCεRI, chymase, tryptase, RMCPI and RMCPII, but were virtually void of other cell markers. Biliary proliferation and fibrosis increased following treatment with MC supernatants from BDL rats + NaCl and these parameters decreased in cells treated with MC supernatants from BDL + cromolyn sodium. In conclusion, we have isolated and characterized MCs from cholestatic livers. MCs regulate cholestatic liver injury and hepatic fibrosis. This tool provides a better understanding of the paracrine influence of mast cells on biliary/liver pathologies.
登录
查看更多内容
影响因子：
24.5
作者：
GITTLEN, SD;SCHULMAN, ES;MADDREY, WC
通讯作者：
MADDREY, WC
影响因子：
5
作者：
Kennedy, Lindsey L.;Hargrove, Laura A.;Francis, Heather L.
通讯作者：
Francis, Heather L.
影响因子：
9.3
作者：
Choi, Joon-Seok;Kim, Jun Ki;Choi, Jin Woo
通讯作者：
Choi, Jin Woo
影响因子：
5
作者：
Koda, W;Harada, K;Nakanuma, Y
通讯作者：
Nakanuma, Y
影响因子：
29.4
作者：
LeSage, G;Glaser, SS;Alpini, G
通讯作者：
Alpini, G