银杏叶提取物对肝星状细胞TGF-β_1,CTGF及细胞外基质表达的影响

Effect of Ginkgo biloba extract on expression of TGF-β_1,CTGF mRNA and extracellular matrix in rat hepatic stellate cells

目的:研究银杏叶提取物对肝星状细胞增殖及转化生长因子(TCF)-β1、结缔组织生长因子(CTCF) mRNA表达及细胞外基质分泌的影响。方法:用不同浓度(0,1,10,100,500 mg·L-1)的银杏叶提取物处理 HSC-T6 24 h及48 h后,用逆转录聚合酶链反应(RT—PCR)法检测各组细胞及空白对照组中TGF—β1,CTGF mRNA的表达;放射免疫法分析上清液中Ⅲ型前胶原、Ⅳ型胶原、透明质酸和层粘连蛋白的含量;MTT及流式细胞仪检测其对肝星状细胞增殖及细胞周期的影响。结果:银杏叶提取物10,100,500 mg·L-1能明显抑制 TGF-β1和CTGF mRNA的表达,降低上清液中Ⅲ型前胶原、Ⅳ型胶原、透明质酸和层粘连蛋白的含量 (P<0．01或P<0．05),在一定范围内呈随剂量增加及时间的延长,抑制效率增加;并可抑制肝星状细胞的增殖,影响HSC-T6的细胞周期,使细胞周期阻滞于G1期。结论:银杏叶提取物可明显抑制HSC-T6的增殖,降低其细胞因子基因表达及细胞外基质的分泌。

AIM： To investigate the effect of Ginkgo biloba extract （GbE761） on hepatic stellate cell proliferation and the expression of TGF-β1,CTGF mRNA and secretion of extracellular matrix in HSC-T6. METH ODS： HSC-T6 were cultured with GbE761 in different concentrations （1,10,100,500 mg·L^-1）and took that without GbE761 as blank control. The expressions of TGF-β1 and CTGF mRNA levels were determined by reverse transcription-polymerase chain reaction （RT-PCR） after 24 h and 48 h. And precollagen type Ⅲ, Ⅳ-type collagen,hyaluronate （HA）,laminin （LN）were determined by radioimmunoassay. MTT colorimetric assay was used for detecting the proliferation of HSCs and flow cytometry was performed for observing the cell cycles of HSCs under different concentrations of GbE761. RESULTS： GbE761 in different concentrations （10,100,500 mg·L^-1）could down-regulate the expressions of TGF-131 and CTGF mRNA and inhibit the secretion of precollagen type Ⅲ,Ⅳ- type collagen, HA and LN in HSC-T6 （P 〈 0.01 or P 〈 0.05） . The effect was increased with a dose and time dependent manner. It could inhibit the proliferation of HSCs with the cell cycle being blocked in G1 phase at the concentration of 10,100 and 500 mg·L^-1. CONCLUSION： GbE761 can exert significant effect on HSC-T6 by inhibiting the proliferation and down-regulating the expression of TGF-β1 and CTGF mRNA, and secretion of extracellular matrix.