Study Of Total Flavone Of Semen Litchi(TFL) On The Nuclear Translocation Of Nuclear Factor-κb And The Expression Of Related Proteins Of Activated Rat Hepatic Stellate Cells In Vitro

Objective: To investigate the effects of total flavone of semen litchi(TFL)on the nuclear translocation of nuclear factor-κB(NF-κB)p65 and the expression of related proteins of activated rat hepatic stellate cells in vitro,and explore the mechanism of anti hepatic fibrosis.Methods: HSC-T6 cells were cultured and stimulated by TGF-β1 for 24 h in vitro,then gave different concentrations of TFL for 48 h.The nuclear translocation of nuclear NF-κB p65 was detected by laser scanning confocal microscopy.The expression levels of TLR4 、 p-NF-κB 、 Bax 、 Bcl-2 and CollagenⅠproteins were measured by western blot.The immunofluorescence was used for observing the expression levels of TLR4 and p-NF-κB proteins.Results:(1)TFL could inhibit the nuclear translocation of NF-κB p65 of activated HSC-T6 in a concentration-dependent manner.(2)The Western blot assay showed that the expression levels of TLR4 protein(0.88 ± 0.054)、p-NF-κB protein(0.79± 0.026)、Bcl-2 protein(0.90± 0.035)、Bax protein(0.287± 0.030)、CollagenⅠprotein(0.825 ± 0.040)in the TGF-β1 group and TLR4 protein(0.7379± 0.031)、p-NF-κB protein(0.6539± 0.028)、Bcl-2 protein(0.725 ± 0.058)、Bax protein(0.4103 ± 0.025)、CollagenⅠprotein(0.704± 0.022)in the control group had obvious difference(P<0.05).And there were no significant differences in the expression of TLR4、p-NF-κB、Bcl-2、Bax and CollagenⅠ protein between control group and TFL125 group(TLR4 protein(0.7895 ± 0.037)、p-NF-κB protein(0.6620 ± 0.0285)、Bcl-2 protein(0.759 ± 0.043)、 Bax protein(0.441 ± 0.025)、CollagenⅠprotein(0.701 ± 0.047)).Compared with the TGF-β1 group,the expression of TLR4、p-NF-κB、Bcl-2、Bax and CollagenⅠ protein of TFL125 group,TFL250 group(TLR4 protein(0.6369 ± 0.020)、p-NF-κB protein(0.5277 ± 0.029)、Bcl-2 protein(0.646 ± 0.048)、Bax protein(0.593± 0.039)、CollagenⅠprotein(0.586± 0.018))and the TFL500 group(TLR4 protein(0.451 ± 0.051)、p-NF-κB protein(0.287 ± 0.036)、Bcl-2 protein(0.486 ± 0.037)、 Bax protein(0.732 ± 0.023)、CollagenⅠprotein(0.364 ± 0.111))had obvious difference(P<0.05).Conclusions: 1.The antifibrosis mechanism of TFL may be by inhibiting the nuclear translocation of NF-κB p65 in activated HSC-T6.2.TFL can reduces the expression of TLR4、p-NF-κB、Bcl-2 and CollagenⅠproteins,increase the expressions of Bax in activated HSC-T6,so as to have the effect of anti-liver fibrosis.3.TFL reduce the expression of TLR4、Bcl-2 and CollagenⅠproteins,increase the expressions of Bax in activated HSC-T6,perhaps by inhibiting NF-κB activity.