应用Alarma blue测定法评价软肝煎药物血清对HSC/T6增殖的影响

目的：应用Alarma blue测定法评价中药复方软肝煎对HSC／T6增殖的影响，方法：实验分为小牛血清，大鼠血清及药物血清3组，各组再分为5％，10％及20％3个浓度组，每组4只大鼠，利用中药血清药理学，通过MTT法和Alarma blue测定法测定软肝煎药物血清影响HSC／T6增殖的情况。结果：在MTT测定法中，与小牛血清组相比，大鼠血清组的5％浓度差异有显著性意义（P＜0．05），10％，20％浓度差异有非常显著性意义（P＜0．01），药物血清组各浓度差异均有非常显著性意义（P＜0．01），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＜0．05），20％浓度差异有非常显著性意义（P＜0．01），在Alarma blue测定法中，24h时，与小牛血清组比较，大鼠血清组与药物血清组的10％浓度差异均有非常显著性意义（P＜0．01），与大鼠血清组相比，药物血清组的10％浓度差异有显著性意义（P＜0．05，）30h时，与小牛血清组比较，大鼠血清组的10％浓度及药物血清组的10％学差异均有显著性意义（P＜0．05，P＜0．01），与大鼠血清组相比，药物血清组的10％浓度差异有非常显著性意义（P＜0．01），结论：软肝煎能抑制HSC／T6的增殖，具有抗肝纤维化作用；Alarma blue测定法是一种可靠的方法，不同动物种属的血清对肝星状细胞增殖影响不同，在培养系中使用单一的血清为好。

Effect of Serum Containing Ruanganjian Decoction on Proliferation of HSC/T6 by Alarma Blue Assay

Objective:To evaluate the effect of serum containing Ruanganjian decoction on the proliferation of HSC/T6 by alarma blue assay.Methods:All of the sera were divided into 3 groups: calf serum group, rat serum group and drug-containing serum group. Each group was subdivided into three subgroups of 5 %, 10 % and 20 % (n=4 each). MTT assay and alarma blue assay were used to measure the effect of Ruanganjian decoction on the proliferation of HSC/T6.Results: In the groups assayed with MTT, rat serum of 5 % could inhibit HSC/T6 proliferation with more remarkable effect than calf serum (P< 0.05), while rat sera of 10 % and 20 % with very significant difference (P < 0.01); There was significant difference among different subgroups of drug-containing serum (P< 0.01); As compared with rat serum, drug-containing sera of 5 % and 10 % could inhibit HSC/T6 proliferation with remarkable difference (P< 0.05), while serum of 20 % with very significant difference (P< 0.01). In the groups assayed with alamar blue, as compared with calf serum, rat and drug-containing sera of 10 % could inhibit HSC/T6 proliferation after 24 h with more significant difference (P< 0.01); As compared with rat serum, drug-containing serum of 10 % could inhibit HSC/T6 proliferation with statistical difference (P< 0.05); As compared with calf serum, rat serum of 10 % could inhibit HSC/T6 proliferation after 30 h with significant difference (P< 0.05), while drug-containing serum of 10 % with more significant difference (P< 0.01); As compared with rat serum, drug-containig serum of 10 % could inhibit HSC/T6 proliferation after 30 h with very significant difference (P< 0.01). Conclusion: (1) Serum containing Ruanganjian decoction could inhibit the proliferation of HSC/T6, antagonize hepatic fibrosis. (2) Alarma blue assay was a creditable method. (3) Different animal sera had different effects on HSC proliferation and it was preferable to use a single animal serum in cultured medium.