软肝煎药物血清对肝星状细胞/T6增殖及Ⅰ型胶原合成的影响

Effects of Serum Containing Ruangan Decoction on Proliferation of HSC/T6 and TypeⅠCollagen Synthesis in Culture

目的 :探讨软肝煎药物血清对肝星状细胞 (HSC) / T6的增殖、 型胶原合成的影响。方法 :实验分为大鼠血清组和药物血清两组 ,各组再分为 5 %、10 %及 2 0 % 3个浓度组 ,每组 4只大鼠。用 MTT比色法和 Alarmablue测定法评价软肝煎药物血清对 HSC/ T6的增殖抑制作用 ;用 Western blotting法和 EL ISA法评价软肝煎药物血清对 HSC/ T6合成 型胶原量的影响。结果 :在 MTT测定法中 ,与大鼠血清组相比 ,药物血清组的 5 %、10 %浓度差异有显著性意义 (P <0 .0 5 ) ,2 0 %浓度差异有非常显著性意义 (P <0 .0 1)。在 Alarma blue测定法中 ,2 4 h时与大鼠血清相比 ,药物血清组的 2 0 %浓度差异有显著性意义 (P<0 .0 5 ) ;30 h时与大鼠血清相比 ,药物血清组的2 0 %浓度差异有非常显著性意义 (P <0 .0 1)。 EL ISA法测定的细胞上清 型胶原含量 ,药物血清组与大鼠血清组相比差异有显著性意义 (P <0 .0 5 ) ,Western blottin法测定细胞内合成 型胶原的量 ,可知药物血清组 型胶原的含量明显少于大鼠血清组。结论 :软肝煎不仅能抑制 HSC的增殖 ,而且能抑制 HSC合成 型胶原的量 ,从而提示软肝煎有很好的抗肝纤维化作用。

Objective:To investigate the effects of serum containing Ruangan decoction on proliferation HSC/T6 and typeⅠcollagen synthesis of in culture. Methods: All of the sera were divided into 2 groups: rat serum group and drug-containing serum group. Each group was subdivided into three subgroups according to the concentration of 5%, 10% and 20% (n=4).MTT spectrometric assay and Alarma blue assay were used to assess the effect of Ruangan decoction on the proliferation of HSC/T6.The amount of typeⅠcollagen secretion was measured by the method of ELISA and that of type Ⅰcollagen synthesis by Western blotting analysis. Results: Serum containing Ruangan decoction could inhibit HSC/T6 proliferation in a concentration-dependent manner. In the groups assayed with MTT spectrometric assay, as compared with rat serum, drug-containing sera of 5%, 10% could restrain the cell proliferation with remarkable difference (P<0.05), while 20% with more significant difference (P<0.01). In the groups assayed with Alamar blue assay, as compared with rat serum, drug-containing serum of 20% could restrain the cell proliferation with statistical difference after 24 hours, (P<0.05); while after 30 hour, 20% could evidently restrain the cell proliferation with more significant difference (P<0.01). The amount of typeⅠcollagen secretion in drug-containing serum of 20% by ELISA assay was remarkably less than that in rat serum (P<0.05). The image of Western blotting analysis showed that the amount of typeⅠcollagen synthesis in drug-containing serum of 20% was also less than that in rat serum group. Conclusion:Serum containing Ruangan decoction could inhibit not only HSC/T6 proliferation, but also typeⅠcollagen synthesis, which indicated that the decoction had good action against hepatic fibrosis.