The influence of regulatory T cells in hepatic stellate cell proliferation and HA secretion

Objective To investigate the effect of CD4+CD25+ regulatory T cells(Treg)in the proliferation of hepatic stellate cells(HSC) and the secretion of Hyaluronic Acid(HA).Methods By using magnetic activated cell sorter(MRCS),Treg cells were isolated from peripheral blood of chronic hepatitis B patients The hepatic stellate cells(HSC)cultured alone were used as blank group.The LX-2 were co-cultured with Treg cells in vitro as co-culture group;adding TGFβ1 to HSC co-cultured as control group.CCK-8 method for the detection of the proliferation of HSC,ELISA analysis for the detection of HA secretion was performed.One-way analysis of variance was used for the comparison.Results Compared with the blank group(0.6623±0.02702) the proliferation of HSC of the co-culture group was significantly higherand the most prominent effect was 2.5∶ 1(0.6836±0.00560),while the proliferation of HSC decreased at 1∶ 1、5∶ 1 and 7.5∶ 1,and the discrepancy between co-culture subgroups and blank group had statistical significance(F=11.421,5.272,84.514,490.639;P0.05).Compared with blank group,the proliferation of HSCs of different concentrations of TGFβ1 control subgroup increased significantly(0.6720±0.00753,0.6773±0.00585,0.5373±0.00387) and reached a peak at 8 ng/μl,and the discrepancy between control subgroups and blank group had statistical significance(F=196.175,799.624,236.461;P0.01).Linear correlation analysis showed concentrations of HA and the proliferation of HSC in co-culture group was positively correlated with control group(r=0.661,0.871;P0.01).Conclusion CD4+CD25+ Treg cells could accelerate the cultivation of HSCs proliferation and HA secretion.And,in vitro,we confirmed CD4+CD25+ Treg cells play an important role in the process of chronic hepatitis B.