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Objective: To investigate hydroxycamptothecin (HCPT) and normal rat liver cell proliferation of cultured rat hepatic stellate cells (HSC), to determine the optimal concentration of HCPT HSC proliferation inhibition; observed HCPT on HSC apoptosis the impact. Lay the experimental basis of cell biology; provide a theoretical basis to broaden the range of clinical applications of camptothecin and experimental basis for further research HCPT hepatic fibrosis. Methods: The rat hepatic stellate cells (HSC-T6) and normal rat liver cell line (BRL-3A) in the experimental group (respectively with HCPT concentration of 0.008, 0.016, 0.031, 0.063, 0.125, 0.25, 0.5 1, 2, 4, 8, 16, 32 mg / L of culture medium role) and the control group (pure culture) in vitro for 24 h, 48 h, 72 h. Application of the the four MTT Act (MTT) cell proliferation was detected to determine the optimal concentration of HCPT inhibition of proliferation of HSC-T6 cells; flow cytometry HCPT apoptosis of HSC-T6; transmission electron microscopy observed under the morphological changes of the HSC-T6 apoptosis; the DNA fragmentation strip apoptosis of cells was observed by agarose gel electrophoresis. RESULTS: MTT method for 24 h, 48 h, the proliferation rate of HSC-T6 cells 72 h experimental group than the control group, the difference was significant (t = 6.07 to 46.98, 10.98 to 63.97, 20.76 to 107.68, P lt HCPT HSC-T6 cells and BRL-3A cell proliferation inhibition rate gradually increased with the higher concentration of drug action and prolonged drug action time;; 0.01); When HCPT concentration gt; 0.5 mg / L The toxic effects of BRL-3A cells significantly increased (all P lt; 0.05). 0.25,0.5,1 mg / L the HCPT role of HSC-T6 cells for 24 h under the light microscope to are visible cells reduce the number, volume reduction, nuclear enrichment and other changes, as the role of concentration increase is more pronounced. 0.125,0.25,0.5 mg / the L the HCPT role of HSC-T6 cells for 24 h, flow cytometry showed that apoptosis rates were: 13.46% ± 2.42%, 26.25% ± 5.65%, 47.05% ± 8.76%, with the control group (4.89% ± 1.80%) compared to the difference was significant (F = 34.24, P lt; 0.01). 0.5mg / L HCPT role in HSC-T6 cells for 24 h under TEM visible cell volume narrow nucleoli disappear, chromatin condensation gathered into lumps along the nuclear membrane arrangement apoptotic morphological changes; agarose gel electrophoresis. seen significant DNA ladder band formation. Conclusion: HCPT in vitro can significantly inhibit the proliferation of HSC-T6 cells, dose intensity - time-dependent; 0.5 mg / L for HCPT on HSC-T6 cells proliferation inhibition concentration. HCPT can also induce apoptosis of HSC-T6 cells, the role of intensity in a dose-dependent manner.
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