| Purpose:The aim of this article is to observe the influence of Rosiglitazone on proliferation, apoptosis, cell circle and ultrastructure of hepatic stellate cells (HSC) in vitro, and also observe the effect of Rosiglitazone on expression ofα-SMA, PDGF-B in HSC cultured respectively in ordinary medium or in conditioned medium of HepG2, And then the mechanism of Rosiglitazone on effect of HSC biological characteristics was discussed.Method:1. MTT assay was used to detect the influence of different concentrations of Rosiglitazone (10μmol/L, 12.5μmol/L, 15μmol/L, 17.5μmol/L, 20μmol/L respectively) on the proliferation of HSC.2. Flow Cytometry (FCM) was employed to investigate the effect of different concentrations of Rosiglitazone on the ratio of apoptosis and the distribution of cell cycle.3. Transmission electron microscope (TEM) was applied to observe the ultrastructural change of HSC after treatment by Rosiglitazone (15μmol/L).4. Immunohistochemistry (IHC): 1) Expression levels ofα-SMA, PDGF-B in HSC treated by different concentrations of Rosiglitazone was semi-quantitative analyzed by IHC; 2) HSC were at first exposed to 24h conditioned medium from HepG2 (1∶1 and 1∶2 dilution) and then treated with Rosiglitazone (15μmol/L), expression levels ofα-SMA, PDGF-B in HSC was also checked by IHC.Results:1. MTT result showed that: Rosiglitazone could inhibit proliferation of the HSC. Inhibition rate increased with increasing Rosiglitazone concentration in a dose-effect relationship.2. FCM result showed that: Both apoptosis rate and the percentage of G0/G1 phase increased with increasing Rosiglitazone concentration. The increase of apoptosis rate in each group was very significant (p<0.01). The increase of percentage of G0/G1 phase was significant (p<0.05), when the concentration of Rosiglitazone was above 15μmol/L.3. TEM result showed that: After Rosiglitazone treatment (15μmol/L), growth state of HSC was inhibited, part of HSC presented the ultrastructural changes of apoptosis, for example, cell volume became small, the number of villi in cell surface was decreased or disappeared, chromatin condensation and margination and apoptotic bodies were observed.4. IHC result showed that: 1) After 24h of treatment with Rosiglitazone, expression ofα-SMA, PDGF-B in HSC was decreased with the increase of Rosiglitazone concentration, expression of both antigen were closely related (r=0.971). 2) HSC were at first exposed to 24h conditioned medium from HepG2 and then treated with Rosiglitazone (15μmol/L), expression ofα-SMA, PDGF-B in HSC was decreased significantly (p<0.05) compared with that in control group and conditioned medium treatment group.Conclusion:Rosiglitazone could inhibit the proliferation and activation of HSC. The mechanism could be related with arresting HSC in the G0/G1 phase, inducing the apoptosis of HSC and decreasing the expression of PDGF in HSC. Rosiglitazone could also inhibit the activation and expression of PDGF in HSC pretreated with conditioned medium of HepG2.
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