【摘要】 目的:研究丹酚酸A对培养的大鼠肝星状细胞增殖与胶原生成的影响。方法:用链酶蛋白酶与胶原酶对肝脏进行原位灌流消化,Nycodenz密度梯度离心分离大鼠肝星状细胞,传一代培养。MTT法与[³H]TdR掺入法测定细胞增殖。丽春红染色、图象分析法半定量细胞胶原沉积量,ELISA法测定细胞培养上清中Ⅰ型胶原分泌量,检测细胞层总蛋白量校正细胞数。RT-PCR法分析前胶原α₂（Ⅰ）基因的表达。结果:丹酚酸A 100μmol/L引起部分细胞脱壁与死亡,有一定毒性反应。丹酚酸A 0.1-10μmol/L对细胞形态无明显影响。丹酚酸A1-100μmol/L浓度依赖性抑制细胞增殖,降低胶原沉积量与Ⅰ型胶原分泌量。丹酚酸A1-10μmol/L对前胶原α₂（Ⅰ）mRNA表达均有明显抑制作用。结论:丹酚酸A抑制肝星状细胞增殖与胶原表达,是丹参抗肝纤维化的主要有效成分之一,抑制肝星状细胞活化是其抗肝纤维化的主要作用机制。更多还原

【Abstract】 To investigate the effects of salvianolic acid-A (SA-A), one of main effective components of Salvia miltiorrhiza for its antifibrotic action, on the cell proliferation and collagen production in cultured hepatic stellate cells (HSC). METHODS: HSC were isolated through in situ perfusion of liver with pronase E and collagenase, and gradient centrifugation with Nycodenz. The cultured HSC were incubated with SA-A 0.1 -100μmol/L for 24 h. MTT spectrometric assay and intercellular incorporation of methyl-[3H]thymidine ([3H]TdR) was used to assess the cell proliferation. The amount of collagen was semi-quantified by ponceau staining and image analysis, the amount of type I collagen secretion was measured with ELISA and normalized by the total protein of cell layer. The total RNA was prepared from the control cells and the drug treated cells respectively, and the expression of pro-collagen α2(Ⅰ) mRNA was semi-quantitatively analyzed with RT-PCR. RESULTS: SA-A 100 μmol/L showed a little cytotoxity, SA-A 0.1 - 10 μmol/L did not influence cell morphology, and SA-A 1 - 100μmol/ L decreased the cell proliferation significantly in a concentration-dependent manner (P < 0.05). SA-A 1, 10, 100 μmol/L decreased the cell collagen deposition by 78.6 %, 71.8 %, and 61.3 % of the control respectively (P <0.05), and decreased type I collagen secretion to 53.1 %, 52.6 %, and 49.5 % (P<0.01 or P <0.05). Both SA-A 1 and 10μmol/L downregulatedprocollagen α2 (Ⅰ) mRNA expression remarkably ( P < 0.05). CONCLUSION: SA-A inhibited HSC proliferation and collagen expression. The inhibitory effect on HSC activation is the main mechanism of SA-A action a-gainst liver fibrosis.更多还原