论文部分内容阅读
目的 探讨丹酚酸B盐 (SA B)抗肝纤维化的作用机理。方法 分离大鼠肝星状细胞(HSC) ,于无包被的塑料培养皿上原代培养 1、4、7d ,细胞分别处于静止、中间活化与完全活化 3种活化状态。予以 10 0pmol/L转化生长因子 β1(TGF β1)刺激 ,观察细胞α 肌动蛋白表达与胶原分泌的变化。选择对TGF β1刺激最为敏感的中间活化状态HSC为细胞模型 ,[3 H]胸腺嘧啶掺入法观察 0 1μmol/L～ 1mmol/LSA B对该细胞增殖的作用 ,倒置显微镜观察药物对细胞形态的影响 ;Western印迹与Northern印迹等方法观察SA B对TGF β1刺激活化的HSC胞外基质基因与蛋白表达、α 肌动蛋白表达的影响 ;提取细胞质与细胞核蛋白 ,Western印迹方法观察SA B对TGF β1刺激的HSCSmad2 /3蛋白表达、磷酸化与核转位的影响。结果 TGF β1促进各活化状态HSC的胶原分泌 ,促进率分别为12 8 6 %、2 0 7 3%与 188 2 % ,中间活化状态HSC对TGF β1的促胶原分泌最为敏感。除 0 1mmol/L～ 1mmol/LSA B引起部分细胞死亡外 ,0 1μmol/L～ 10 μmol/LSA B对细胞形态无影响 ,但可浓度依赖性地抑制细胞增殖 ,细胞内 [3 H]胸腺嘧啶掺入量分别为对照组的 76 %、6 0 1%与 4 7 8%。 1μmol/L～ 10μmol/LSA B明显抑制TGF β1刺激的HSC胶原分泌量 (分别为对照组的
Objective To explore the mechanism of salvianolic acid B salt (SA B) in the treatment of liver fibrosis. METHODS Rat hepatic stellate cells (HSCs) were isolated and cultured on uncoated plastic plates for 1, 4, and 7 days. The cells were activated, activated, and fully activated. The cells were stimulated with 100 μmol/L transforming growth factor β1 (TGF β1) and the changes of α-actin expression and collagen secretion were observed. Select the intermediate activation state HSC most sensitive to TGF β1 stimulation as a cell model, and observe the effects of 0 1μmol/L～1mmol/LSA B on the proliferation of HSC by [3 H]thymidine incorporation method. Observe the effect of drugs on cell morphology by inverted microscope. Effect; Western blotting and Northern blotting were used to observe the effect of SA B on TGF β1 stimulated expression of extracellular matrix gene and protein and α-actin in activated HSC; cytoplasm and nuclear protein were extracted; Western Blot was used to observe SA B on TGF β1. Stimulation of HSCSmad2/3 Protein Expression, Phosphorylation and Nuclear Translocation Results TGFβ1 promoted the secretion of collagen in activated HSCs with the rates of 128.6 %, 207 3% and 188 2 % respectively. The intermediate activation state of HSC was most sensitive to the secretion of TGF β1. Except for 0 1mmol/L～1mmol/LSA B causing partial cell death, 0 1μmol/L～10μmol/LSA B had no effect on cell morphology, but could inhibit cell proliferation in a concentration-dependent manner, intracellular [3 H]thymidine The incorporation amount was 76%, 601%, and 47.8% of the control group, respectively. 1μmol/L～10μmol/LSA B significantly inhibited TGFβ1-stimulated HSC collagen secretion (controls, respectively