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AIM:To investigate the effects of IH764-3 on HSC apoptosis,and the expression of caspase-3 protein in HSC apoptoticprocess.METHODS:HSCs were cultured in medium with differentIH764-3 doses(10μg·mL~(-1),20μg·mL~(-1),30μg·mL~(-1),40μg·mL~(-1))and without IH764-3,and HSC proliferation wasquantitatively measured by ~3H-thymidine incorporation.Themorphological changes of HSCs were observed withtransmission electron microscope after exposure to the doseof 40μg·mL~(-1)of IH764-3 for 48 hr.The apoptosis rates weredetected by annexin V/PI and TdT-mediated dUTP nick endlabeling(TUNEL).The expression of caspase-3 protein wasdetermined by flow cytometry.RESULTS:(1)HSC proliferation rates induced with different1H764-3 doses(10μg·mL~(-1),20μg·mL~(-1),30μg·mL~(-1),40μg·ml~(-1))were significantly reduced compared with that of thecontrol group(P<0.01).(2)With the doses above,IH764-3dose-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 csspase-3 protein in 11-1764-3 groups wassignificantly 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 HSCapoptosis.Furthermore,IH764-3 can significantly increasethe caspase-3 protein expression.
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), 40μg·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 40μg·mL -1 for IH764-3 for 48 hr.The apoptosis rates weredetected by annexin V/PI and TdT-mediated dUTP nick endlabeling(TUNEL).The expression of caspase-3 protein Wasdetermined by flow cytometry.RESULTS: (1) HSC proliferation rates induced with different1H764-3 doses (10 μg·mL -1 ), 20 μg·mL -1, 30 μg·mL -1, 40 μg·ml ~(-1))were less 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%) respe Ctively, 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 csspase-3 protein in 11-1764-3 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 HSCapoptosis.Furthermore,IH764-3 can significantly increasethe caspase-3 protein expression.