The role of myosin light chain kinase in intestinal epithelial barrier dysfunction due to hypoxia.
- Author:
Pei WANG
1
;
Chuan-li CHEN
;
Mu LI
;
Feng-jun WANG
Author Information
- Publication Type:Journal Article
- MeSH: Caco-2 Cells; Epithelium; metabolism; physiopathology; Humans; Hypoxia; metabolism; physiopathology; Intestinal Absorption; Intestinal Mucosa; metabolism; physiopathology; Intestines; cytology; metabolism; physiopathology; Myosin Light Chains; metabolism; Myosin-Light-Chain Kinase; metabolism
- From: Chinese Journal of Burns 2009;25(1):57-60
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the role of myosin light chain kinase (MLCK) in intestinal epithelial barrier dysfunction after hypoxia.
METHODSThe Caco-2 monolayers developed with Transwell inserts were exposed to hypoxia for 0 h (NC group), 2, 6, 8, 12 and 24 h (H group), and 6 h hypoxic specimens were treated with 100 mol/L ML-9 (T group). The transepithelial electrical resistance (TER) of monolayers was measured with an ohmmeter. The tight junction protein ZO-1 of monolayers was analyzed by immunofluorescence assay. The protein expressions of phosphorylated myosin light chain (p-MLC) and MLCK were detected by Western blotting.
RESULTSThe TER of monolayers in H group at 6, 8, 12 and 24 h was 422 +/- 17, 427 +/- 27, 403 +/- 40 and 426 +/- 22 ohms respectively, which was significantly lower than that of NC group (451 +/- 27 ohms, P < 0.05). The TER of monolayers in T group was 558 +/- 110 ohms, which was significantly higher than that in H group at each time point ( P < 0.01). The ZO-1 of monolayers in H group at 6 h was irregular in arrangement, with interruptions and rugae, and sawtooth. These abnormalities were ameliorated in T group (regular in arrangement, with little or without ruga and sawtooth). The protein expressions of p-MLC and MLCK in H group at each time point were higher than those in NC group.
CONCLUSIONSIntestinal epithelial barrier dysfunction after hypoxia can be mediated by MLCK.
