Differential proteomic analysis of rat hepatic stellate cells treated by oxymatrine liposomes using two-dimensional electrophoresis.
- Author:
Ning-Li CHAI
1
;
Qing CHANG
;
Shi-Ping XU
;
Jun WAN
;
Ben-yan WU
Author Information
- Publication Type:Journal Article
- MeSH: Alkaloids; pharmacology; Animals; Electrophoresis, Gel, Two-Dimensional; Hepatic Stellate Cells; cytology; drug effects; metabolism; Liposomes; Liver Cirrhosis, Experimental; metabolism; Male; Proteome; metabolism; Quinolizines; pharmacology; Rats; Rats, Sprague-Dawley
- From: Chinese Journal of Integrated Traditional and Western Medicine 2013;33(5):679-685
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo analyze differentially expressed proteins of hepatic stellate cells (HSCs) treated with oxymatrine (OMT) liposomes, thus further exploring the molecular mechanism of OMT liposomes for treating liver fibrosis.
METHODSA rat model of CCl4 induced chronic liver fibrosis was established. HSCs were perfusion isolated from modeled SD rats and cultured in vitro . Passage 2 HSCs were divided into the model group (Group A), the OMT-liposome-treated group (Group B), and the liposome-treated control group (Group C). HSCs from normal rats were taken as the normal control group (Group D). The total proteins of HSCs cells were extracted from Group B and D after 7 days of treatment, and separated with isoelectrofocusing two-dimensional electrophoresis (2-DE). A 2-DE system was established to analyze the differences in the protein profile between Group B and Group C. Tow protein dots with most obvious difference were selected to determine the structures and functions of different proteins using peptide mass fingerprinting (PMF).
RESULTS(1) The total number bf proteins decreased after treated with OMT liposomes, with 864 spots before treatment and 756 spots after treatment, and the matching rate was 63%. (2) According to 2-DE results, 10 differential protein spots were found by image analysis of magnifying images in local regions. (3) Two most differently expressed proteins were identified to be ATM (46. 236 kD) and Miz1 (54. 051 kD) by PMF and SWISS-PROT protein database retrieval.
CONCLUSIONAction of OMT liposomes on HSCs of rats with chronic liver fibrosis caused different protein expressions, which might be involved in the signaling pathways of inducing the apoptosis of HSCs.