Objective      To explore the effect of serum containing modified turtle shell decoction (MTSD) on the proliferation

cell cycle and apoptosis of hepatic stellate cell line T6 (HSC-T6).       Methods      Serum containing MTSD or turtle shell pills (TSP) was prepared

and the rats were treated with MTSD at 28.4 g/(kg·d) as a high-dose group

at 14.2 g/(kg·d) as a medium-dose group

at 7.1 g/(kg·d) as a low-dose group or with TSP at 9.0 g/(kg·d) as a control group by intragastric administration. HSC-T6 cells were treated with serum containing different doses of MTSD in equal to volume. The same cells were treated with serum containing TSP as a positive control

treated with normal salt solution as a blank control

and treated with normal rat serum in equal to volume as a normal control. The proliferation of HSC-T6 cells in different groups were tested by cell count kit-8 (CCK-8) assay

and the proliferation curves were draw. Cell cycle of HSC-T6 cells was tested by flow cytometry (FCM). Annexin V-PI was applied to analyze the apoptosis of the cells. The gene and protein expressions of proliferating cell nuclear antigen (PCNA) of HSC-T6 cells were detected with RT-PCR and Western blot

respectively.       Results      The results of CCK-8 assay revealed that serum containing MTSD could prevent HSC-T6 cells from proliferation with a dose-effect relationship. The proliferation of the HSC-T6 cells treated with high-dose or middle-dose serum containing MTSD was lower than that of the controls

and the differences were statistically significant after the cells were treated for 48 h (P0.05

P0.01

respectively). Serum containing MTSD affected the cell cycle of HSC-T6 cells

and blocked the cells at S stage. Serum containing MTSD could significantly induce apoptosis of HSC-T6 cells compared with the controls (P0.01). The results of RT-PCR

Western blot and quantitative analyses indicated that the high-dose or middle-dose serum containing MTSD significantly down-regulated the gene and protein expressions of PCNA of HSC-T6 cells

and the differences between the treated cells and the controls were statistically significant (P0.05

P0.01

respectively).       Conclusion      The serum containing MTSD can significantly inhibit proliferation of HSC-T6 cells

induce cell apoptosis and down-regulate the PCNA expressions.