Published online Jun 15, 2002. doi: 10.3748/wjg.v8.i3.515
Revised: January 3, 2002
Accepted: January 23, 2002
Published online: June 15, 2002
AIM: 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 μg·mL-1, 20 μg·mL-1, 30 μg·mL-1, 40 μg·mL-1) and without IH764-3, and HSC proliferation was quantitatively measured by 3H-thymidine incorporation. The morphological changes of HSCs were observed with transmission electron microscope after exposure to the dose of 40 μg·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 μg·mL-1, 20 μg·mL-1, 30 μg·mL-1, 40 μg·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).
CONCLUSION: 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.