阿魏酸钠对转化生长因子β_1抑制肝细胞增殖作用的影响

Effects of sodium ferulate on hepatocyte growth inhibited by transforming growth factor β_1

目的 探讨转化生长因子 β1(TGFβ1)抑制肝细胞生长过程中 ,阿魏酸钠 (sodiumferulic,SF)的干预作用。方法 以培养的正常人肝细胞株 (L0 2细胞 )为对照 ,5 μg·L-1TGFβ1处理的细胞为模型组 ,MTT比色法及3 H TdR参入法分别检测TGFβ1及SF对L0 2细胞生长和DNA合成的影响 ;以流式细胞计数仪分析TGFβ1和SF对L0 2细胞周期各时相变化的影响 ;采用化学荧光探针DCFH DA ,经流式细胞计数仪检测TGFβ1致细胞内活性氧自由基 (ROS)生成情况及不同浓度的SF的干预作用。结果 5 μg·L-1TGFβ1处理L0 2细胞 2 4h ,细胞存活率下降为对照组的 6 7 93% (P <0 0 1vs对照组 ) ,细胞DNA合成抑制率为 37 5 6 % ,2 5 0μmol·L-1SF可以使L0 2细胞存活率升高 19 2 8% (P <0 0 1vs模型组 ) ,DNA合成增加 17 71% (P <0 0 5vs模型组 ) ;SF促进L0 2细胞周期由G0 /G1期向S期和G2 /M期过渡 ,12 5 μmol·L-1和 2 5 0 μmol·L-1SF使L0 2细胞周期S期的比例分别增加 11 4 5 %和 17 92 % (P <0 0 1vs模型组 ) ;5 μg·L-1TGFβ1使L0 2细胞内ROS产生量升高为对照组的1 90倍 ,6 2 5 μmol·L-1～ 2 5 0 μmol·L-1SF分别降低TGFβ1的诱导作用 14 5 %～ 2 7 5 %。结论 阿魏酸钠对抗由TGFβ1引起的肝细胞生长抑制作用 。

AIM To research effects of sodium ferulate(SF) on transforming growth factor β_1 inhibited hepatocyte growth. METHODS Human cultured hepatocytes(L02 cells) served as control group ,TGFβ_1(5 μg·L -1 ) was used to inhibited L02 cells to construct the cell model. MTT methods were used to study the growth inhibitory effect of TGFβ_1 on L02 cells. DNA synthesis was analyzed by measuring 3H-TdR incorporation. The effects of SF on the changes of cell cycle were analyzed by flow cytometry. Generation of intracellular ROS was determined by the emission of fluorescence in L02 cells proloaded with DCFH-DA fluoresin and then treated them with TGFβ_1 or SF. After 4h,control, TGFβ_1 and TGFβ_1+SF-treated cells were analyzed by flow cytometry. RESULTS ①Incubated in the presence of TGFβ_1 for 24 h, 67.93% of L02 cells survived (P<0.05 vs control).250 μmol·L -1 of SF increased cell viability by 19.28% (P<0.05 vs treatment),and the 3H-TdR incorporation reduced by TGFβ_1 by 17.71% (P<0.05 vs TGFβ_1 group). ② 125 μmol·L -1 and 250 μmol·L -1 of SF increased the percentage of cells in S phase by 11.45% and 17.92% (P<0.01 vs control), in G_2 phase by 3.29% and 7.85% respectively(P<0.05 vs control). ③ By treating L02 cells with TGFβ_1 for 4 h, the level of intracellular peroxide was 1.90-fold compared with that of untreated cells. 62.5 μmol·L -1 ～250 μmol·L -1 SF obviously decreased fluorescence by 14.5% to 27.5% respectively. CONCLUSION SF could reduce TGFβ_1 induced growth arrest of L02 cells, which may be associated with the antioxidant effect and the promotion of the cell cycle processes from G_1 phase to S phase.-