The Effect Of Biejiajian Pill Different Extracts On TGFβ1-induced HSC-T6Cells

Objective:To investigate the effects of the Biejiajian pill different extracts on the proliferation and expression of cytokines of TGFβ1-induced HSC-T6cell, and to explore the similar and different anti-hepatic fibrosis functions of aqueous extract and ethanol extracts.Methods:TGFβ1was used to induce HSC-T6cell to establish the proliferation model. With colchicine as a positive control, cells were divided into blank group, colchicine group, the aqueous extract group and ethanol extract group. MTT assay was applied to observe the dose-effect and time-effect of extracts on HSC-T6cell proliferation, and to explore the effect of Biejiajian pill extracts on TGFβ1-induced HSC-T6cell. A fluorescence RT-PCR assay was used to measure the effect of Biejiajian pill extracts on expression of COL-Ⅲ, COL-IV, a-SMA, CTGF, PDGF mRNA in HSC-T6cell. ELISA assay was used to detect the influence of Biejiajian pill extracts on the contenting of COL-Ⅲ, COL-IV, a-SMA, CTGF, PDGF in HSC-T6.Results:1、 MTT assay results showed that it could significantly promote the HSC-T6cell proliferation under the condition of serum free medium in5ng/ml concentration TGFβ1for24h (P<0.05). Both aqueous extract and ethanol extract had an inhibitory effect on the proliferation of TGFβ1-induced HSC-T6cell, while their inhibition performances had certain differences. Aqueous extract in0.125mg/ml concentration for24h had optimal inhibitory effect on HSC-T6proliferation (P>0.05), and there was significant negative linear correlation between dose and inhibition effect in the0.125-2.000mg/ml concentration range with correlation index0.8768. Ethanol extract had no significant negative linear correlation between dose and inhibition effect in the0.125-2.000mg/ml concentration range, but time dependence.2、 RT-PCR results showed that compared with the blank control group, model group increased the content of COL-Ⅲ, COL-IV, a-SMA, CTGF, PDGF mRNA. Compared with the model group, aqueous extract reduced the content of COL-Ⅲ, COL-Ⅳ, a-SMA, CTGF, PDGF mRNA. Ethanol extract reduced the content of COL-Ⅲ, COL-IV, but increased PDGF mRNA.3、 ELISA results showed that compared with the model group, aqueous extract inhibited the expression of COL-Ⅲ, COL-IV, PDGF. Ethanol extract inhibited the expression of COL-Ⅲ, COL-IV, but promoted expression of PDGF.Conclusion:1、 Stability cell activation model of HSC cell is established under the condition of serum free medium in5ng/ml concentration TGFβ1for24h.2、 Both aqueous extract and ethanol extract of Biejiajian pill have an inhibitory effect on the proliferation of TGFβ1-induced HSC-T6cell. For aqueous extract, there was significant negative linear correlation between dose and inhibition effect in the0.125-2.000mg/ml concentration range. For ethanol extract, the inhibitory effect was dependent on time,3、 Combined with RT-PCR and ELISA results that both aqueous extract and ethanol extract of Biejiajian pill can inhibit expressions of COL-Ⅲ and COL-IV. For aqueous extract, it may inhibit the proliferation of HSC-T6by inhibiting the expression of PDGF. For ethanol extract, it may inhibit the proliferation of HSC-T6by promoting the expression of PDGF.