The Bugan Bulk Serum Containing Rat Hepatic Stellate Cell Membrane Pbr Expression

Objectives: To explore Bugansan medicated serum on hepatic stellate cells (HSC) proliferation, cell cycle and apoptosis, along with the discussion from PBR aspect, so as to provide novel theoretical basis and drug source for prevention and treatment for hepatic fibrosis with Chinese traditional medicine.Methods: 5%, 10% and 20% Bugansan medicated serum were prepared as culture medium for Treatment Group, and served as contrast to Control Group that contained no rat serum and Bugansan free culture medium. After HSC-T₆ cell lines recovering and subcultured with Bugansan medicated serum of different concentration, cell proliferation was measured by MTT method, absorption value was determined on an enzyme-labelling instrument and the inhibition rate of Bugansan medicated serum of different concentration on cell proliferation was calculated, meantime, the cell cycle was detected and proportion of cells in different cycle stage was determined using flow cytometry (FCM) to reflex the cell cycle process and proliferation. In different stages of PBR expression, apoptotic index was determined by FCM. And simultaneously radioimmunoassay was applied for detecting binding capacity of PBR specific ligand [³H]PK₁₁₁₉₅ to discuss the relation of cell apoptosis to PBR.Results: 1. Very obvious difference lied in comparison of inhibition rate to HSC proliferation between Bugansan medicated serum groups and Control group(P<0.01), among which the best effectiveness was in 20% concentration medicated serum group with the strongest effect on inhibiting proliferation of HSC; and the group that less better was that of 10%. The least effectiveness appeared in that of 5% group. No statistical significance was observed in comparison between serum free group and control group(P> 0.05). 2. Proportion of G₀/G₁ cell increased corresponded to the increased concentration of Bugansan medicated serum, along with a gradual decline of cells in S stage as well as that in G₂/M stage. All groups of Bugansan medicated serum were statistically significant as compared with control group, whereas non-significance lied in comparisons among serum contained groups (P> 0.05). This indicated a slow process of HSC cycle caused by Bugansan medicated serum, keeping the cells silent in G₀/G₁ stage.3. In every stage of HSC culture, apoptotic index increased obviously with the increasing concentration of Bugansan medicated serum. A superior effect of 10% and 20% medicated serum groups to that of 5% group was observed (P<0.01). At the 11^th day (1 day after subculture) and 13^th day (3 days after subculture), 20% medicated serum group was obvious more effective than that of 10% group(P< 0.05); while at 15^th day (5 days after subculture) and 17^th day (7 days after subculture), there were no statistical significance(P> 0.05).4. At all stages during HSC culture, binding capacity of HSC membrane with [³H]PK₁₁₁₉₅ decreased obviously with the increasing concentration of Bugansan medicated serum. A superior effect of 10% and 20% medicated serum groups to that of 5% group was observed (P<0.01). At the 11^th day (1 day after subculture) and 13^th day (3 days after subculture), 20% medicated serum group was obvious more effective than that of 10% group(P<0.05); and at 15^th day (5 days after subculture), there were no statistical significance(P> 0.05); while at 17^th day (7 days after subculture), 20% medicated serum group showed an obvious greater efficacy than that of 10% group (P< 0.05).Conclusions: Bugansan medicated serum is effective in effecting the life cycle of HSC, inhibiting its proliferation and accelerating its apoptosis. In addition, Bugansan medicated serum promotes cell apoptosis via PBR pathway by intervening or blocking PBR from combining to functionalligand.