龙血素对肝星状细胞的影响

Influence of treatment with loureirin on rat hepatic stellate cells in vitro

目的:观察龙血素A/B对肝星状细胞(hepatic stellate cells,HSC)增殖及各种物质表达的影响.方法:将HSC分为3组:空白对照组、细胞对照组、药物干预组.用MTT法观察龙血素A/B对HSC-T6增殖的影响,并检测细胞上清中透明质酸酶(hyaluronidase,HA)、层粘连蛋白(hyaluronidase,LN)、Ⅳ型胶原(collegen type Ⅳ,Ⅳ-C)的含量.采用Real time-PCR检测HSC经龙血素A/B处理前后血管内皮生长因子(blood vessel endothelium,VEGF165)和低氧诱导因子(histoplasma tissue inhibitory factor,HIF-1)mRNA表达的影响.结果:HSC的抑制率均随龙血素A/B的浓度增高而增高,且龙血素A的IC50约为0.3g/L,龙血素B的IC50约为0.1g/L;与正常对照组相比,药物干预组的HA、LN、Ⅳ-C含量降低(31.10±4.32vs43.05±4.96,441.28±25.38vs302.98±29.59,17.96±3.00vs25.23±4.96,均P<0.05);VEGF和HIF-12-△△Ct值明显降低,且龙血素B组较龙血素A明显.结论:龙血素A/B均可抑制HSC的增值,龙血素A抑制较龙血素B明显;其还可下调HSC中HA、LN、Ⅳ-C及VEGF165和HIF-1mRNA的表达,龙血素B较龙血素A下调效果明显.

AIM：To investigate the effects of treatment with loureirin A/B on rat hepatic stellate cells（HSCs） in vitro.METHODS：HSC-T6 cells were cultured in 96-well plates for 24 h and then incubated with different concentrations of loureirin A/B for 48 h.MTT assay was used to assess the proliferation of HSCs.The contents of hyaluronic acid,laminin and collagen type IV in culture supernatants were measured by radioimmunoassay.The mRNA expression of blood vessel endothelium（VEGF 165） and histoplasma tissue inhibitory factor（HIF-1） was detected by real-time fluorescence quantitative PCR.RESULTS：The half maximal inhibitory concentrations（IC 50） of loureirin A/B were 0.3 and 0.1 g/L,respectively.Treatment with loureirin A/B significantly inhibited HSC proliferation,the secretion of hyaluronic acid,laminin and collagen type IV（31.1±4.32 vs 43.05±4.96,441.28±25.38 vs 302.98±29.59,17.96±3.00 vs 25.23±4.96,all P0.05）,and the mRNA expression of VEGF 165 and HIF-1.The effects of loureirin B were stronger than those of loureirin A.CONCLUSION：Treatment with loureirin A/B could significantly inhibit the proliferation of HSCs,the secretion of hyaluronic acid,laminin and collagen type IV,and the mRNA expression of VEGF 165 and HIF-1,and the effects of loureirin B were stronger than those of loureirin A.