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为探讨 TGFβ1 反义寡核苷酸 (antisense oligonucleotides,ASON)对大鼠肝脏星状细胞活化及其胶原生成的作用 ,作者合成了特异性的 TGFβ1 硫代磷酸 ASON及其对照 (正义 ,错义寡核苷酸 ) ,并将其加入至培养的肝脏星状细胞中。通过细胞免疫化学方法分析 α- SMA的表达来观察肝脏星状细胞的活化 ,肝脏星状细胞产生TGFβ的水平用生物学方法测定 ,以 3H-脯氨酸掺入抑制率的测定观察胶原生成的变化。结果 :针对 TGFβ1 m RNA转译起始区的 ASON在终浓度为 1 0 μm ol/ L 时 ,能明显地抑制体外培养的肝脏星状细胞的激活、TGFβ1 的产生及其胶原的生成能力 ,而对照组寡核苷酸在相同浓度下未见抑制效应。上述结果表明 ,ASON对肝脏星状细胞无损伤作用。 TGFβ1 ASON可能成为一种潜在的抗肝纤维化治疗的药物。
To investigate the effect of TGFβ1 antisense oligonucleotides (ASON) on hepatic stellate cell activation and collagen production in rats, we synthesized a specific TGFβ1 phosphorothioate ASON and its control (sense, Nucleotides) and added to cultured hepatic stellate cells. The expression of α-SMA was analyzed by immunocytochemistry to observe the activation of hepatic stellate cells, the level of TGFβ produced by hepatic stellate cells was determined by a biological method, and the inhibition of 3H-proline incorporation was measured to observe the effect of collagen production Variety. Results: ASON targeting TGFβ1 m RNA translation initiation region inhibited the activation of hepatic stellate cells, TGFβ1 production and collagen production in vitro at a final concentration of 10 μmol / L, while the control Group of oligonucleotides in the same concentration no inhibitory effect. The above results show that ASON has no damage on hepatic stellate cells. TGFβ1 ASON may be a potential anti-hepatic fibrosis drug.