【摘要】 目的观察大黄虫丸经旁分泌途径对大鼠肝星状细胞增殖及其表达TGF-β1基因的影响。方法采用Nycodenz分离液非连续密度梯度离心法同步分离急性CCl4损伤模型大鼠和正常大鼠肝的枯否细胞(KC)和星状细胞(HSC)。在制备正常鼠血清和大黄虫丸药物血清的基础上,将它们分别加入KC培养板内作用48 h后制备成正常鼠KC条件培养液(NKCCM)和损伤鼠KC条件培养液(KCCM)。将NKCCM和KCCM作用于活化的HSC 24 h后,采用MTT法和3H-TdR掺入法检测HSC增殖的变化;采用半定量RT-PCR法扩增TGF-β1mRNA以检测HSC表达TGF-β1基因的情况。结果旁分泌组各组的HSC增殖幅度及TGF-β1基因的相对表达量较正常对照组NKCCM组均明显增加(P<0.05或0.01)。同时旁分泌组中,含药血清作用的5%,10%和20%KCCM组较无药物血清作用的0%KCCM组HSC的增殖能力及TGF-β1基因的相对表达量则下降,且随药物浓度的升高逐渐降低,其组间差别有统计学意义(P<0.05或P<0.01)。结论大黄虫丸可明显抑制经旁分泌途径活化的大鼠HSC的增殖及TGF-β1基因的表达,且其抗肝纤维化效应与药物剂量间存在一定的依赖关系。更多还原

【Abstract】 Objective To observe the effects of Dahuang Zhechong Pill on the proliferation of rat hepatic stellate cells(HSCs) activated by paracrine pathway and the expression of transforming growth factor-beta 1(TGF-β1) gene.Methods The CCl4-injured and normal rat Kupffer cells(KCs),hepatic stellate cells were separated by non-continuous density gradient centrifugation of Nycodenz medium.After the common rat and the drug sera prepared,they were added respectively into Kupffer cell culture plates for normal Kupffer cells conditioned medium(NKCCM) and injured rat Kupffer cells conditioned medium(KCCM) after 48 h.We observed the changes of proliferative HSCs by the methods of MTT and 3H-thymidine(3H-TdR) incorporation assay,and the expression of TGF-β1 gene in HSCs by semi-quantitative RT-PCR when HSCs were actived for 24 h by those conditioned medium.Results In all paracrine groups,the multiplication of HSCs and the relative expression of TGF-β1 gene were increased markedly compared to normal control group(P<0.05 or P<0.01),and gradually decreased in all drug and no drug serum groups with elevated drug concentration.There had significant diferences among all groups.Conclusion Dahuang Zhechong Pill can obviously inhibit the proliferation of rat HSCs activated by paracrine pathway and the expression of TGF-β1 gene in HSCs and deerease hepatic fibrosis in a concentration-dependent manner.更多还原