摘要
目的探讨丹芍化纤方抗肝纤维化作用的机制。方法采用Ⅳ型胶原酶消化及Ficoll密度梯度离心的方法,分离、培养大鼠肝细胞,先加入5%、10%和20%药物血清干预体外培养的肝细胞24h,再用四氯化碳(CCl4)蒸熏法制造肝细胞损伤模型。分别用MTT法及荧光实时定量PT-PCR(RealTimeRT-PCR)法检测CCl4损伤肝细胞的增殖、天冬氨酸特异的半胱氨酸蛋白酶-3(cysteinylaspartate-specificprotease,cas-pase-3)mRNA表达,并检测细胞培养上清中的谷草转氨酶(aspartatetransaminase,AST)活性。结果丹芍化纤方药物血清能促进损伤肝细胞的增殖,下调肝细胞内caspase-3mRNA的表达,降低培养上清中的AST活性,与正常大鼠血清比较差异有显著性,呈明显的血清浓度依赖关系。结论丹芍化纤方抗肝纤维化机制与其抑制肝细胞凋亡、保护肝细胞免受损害有关。
[Objective] To evaluate the antifibrostic mechanism of DanShao HuaXian（ DSHX ） Recipe on hver fibrosis of rats. [Methods] Hepatocyte（HC） was isolated by Ficoll density gradient centrifuge after digest in collagenase Ⅳ, then 5% 10% 20% corresponding serum were added orderly to interfere HC and HC were injured by CCl4 in Vitro. MTT was used to examine the proliferation of HC, the expression of caspase-3 mRNA was detected by real time RT-PCR, and the AST of supernatant was examined. [Results] DSHX-containing serum significantly promoted the proliferation and the AST activity in the cultural medium of HC, downregnlated the expressions of caspase-3 mRNA. There was remarkable differerce as compared with control serum.lts were drug concentration-dependent relations. [Conclusion] Antifibrotic mechanisms of DSHX may be related to its preventing the injury and restraining the apoptosis of HC.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2005年第23期3559-3562,3565,共5页
China Journal of Modern Medicine
基金
贵州省高层次人才基金
编号:2003-1
