Salvia miltiorrhiza monomer IH764-3 induces hepatic stellate cell apoptosis via caspase-3 activation.
OBJECTIVE
To investigate the effects of IH764-3 on HSC apoptosis and the expression of caspase-3 protein in HSC apoptotic process.


METHODS
HSCs were cultured in medium with different IH764-3 doses(10 microg.mL(-1) 20 microg.mL(-1) 30 microg.mL(-1) 40 microg.mL(-1)) and without IH764-3 and HSC proliferation was quantitatively measured by (3)H-thymidine incorporation. The morphological changes of HSCs were observed with transmission electron microscope after exposure to the dose of 40 microg.mL(-1) of IH764-3 for 48 hr. The apoptosis rates were detected by annexin V/PI and TdT-mediated dUTP nick end labeling (TUNEL). The expression of caspase-3 protein was determined by flow cytometry.


RESULTS
(1) HSC proliferation rates induced with different IH764-3 doses (10 microg.mL(-1) 20 microg.mL(-1) 30 microg.mL(-1) 40 microg.mL(-1)) were significantly reduced compared with that of the control group (P<0.01). (2)With the doses above,IH764-3 dose-dependently produced HSC apoptosis rates of 6.7%(9.4%) 9.3%(21.6%) 15.1%(27.2%) and 19.0%(28.4%) respectively by annexin V and PI-labeled flow cytometry assay or TUNEL while it was only 2.3%(6.7%) in the control. (3) The expression of caspase-3 protein in IH764-3 groups was significantly higher than that of the control (P<0.05).


CONCLUSIONS
Within the dose range used in present study IH764-3 can inhibit HSC proliferation as well as enhance HSC apoptosis. Furthermore IH764-3 can significantly increase the caspase-3 protein expression.
Zhang
Xiao-Lan
XL
Department of Gastroenterology The Second Hospital of Hebei Medical University Shijiazhuang 050000 Hebei Province China.
Liu
Li
L
Jiang
Hui-Qing
HQ
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects*; D008103:Liver Cirrhosis / Q000188:drug therapy; D008517:Phytotherapy; D051381:Rats; D027561:Salvia miltiorrhiza
D004365:Drugs, Chinese Herbal; C079053:IH 764-3; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases