丹参注射液对人胚肺成纤维细胞增殖、凋亡及羟脯氨酸表达的影响

Effect of Salvia miltiorrhiza injection on proliferation, apoptosis and expression of amino acid in human embryonic lung fibroblast cells

目的:探讨丹参注射液对人胚肺成纤维细胞增殖、凋亡及羟脯氨酸表达的影响。方法:转化生长因子β1(TGF-β1)诱导人胚肺成纤维细胞增殖;四甲基偶氮唑蓝法检测丹参注射液对人胚肺成纤维细胞数的影响;免疫细胞化学法检测增殖细胞核抗原表达;流式细胞仪检测细胞凋亡;比色法观察羟脯氨酸表达。结果:(1)TGF-β1(0、2.5、5、10、20、40μg/L)可剂量依赖性地上调人胚肺成纤维细胞的细胞数。丹参注射液可剂量依赖性下调人胚肺成纤维细胞光密度值。(2)TGF-β1(10μg/L)干预人胚肺成纤维细胞24、48 h时,增殖细胞核抗原表达明显上调,与同时间点对照组相比差异有统计学意义(P<0.01)。但丹参注射液预处理后,未发现增殖细胞核抗原表达下调。(3)流式细胞仪检测结果显示,丹参注射液预处理后,与TGF-β1组相比,细胞凋亡率增加(P<0.05)。(4)丹参注射液可下调人胚肺成纤维细胞羟脯氨酸表达,与TGF-β1组相比差异有统计学意义(P<0.01)。结论:丹参注射液可靶向人胚肺成纤维细胞,通过促进其凋亡、下调羟脯氨酸表达影响气道重塑。

Objective To investigate the effect of Salvia mihiorrhiza injection on proliferation, apoptosis and expression of hydroxyproline in human embryonic lung fibroblast cells. Methods TGF-β1 was administered to induce the proliferation in human embryonic lung fibroblast ceils; the effect of Salvia miltiorrhiza injection on the number of human embryonic lung fibroblast cells was detected by MTT method; ki67 expression by immunocytochemical method;cell apoptosis by flow cytometry and the expression of hydroxyproline by colorimetry method. Results TGF-β1 （0, 2.5, 5, 10, 20, 40 μg/L）could up-regulate cell number of human embryonic lung fibroblast cells in a dose-dependent manner, while the OD value of human embryo lung fibroblasts cells declined pretreated with Salvia miltiorrhiza injection in a dose-dependent manner and Salvia mihiorrhiza injection could induce the apoptosis and down regulated hydroxyproline expression in human embryo lung fibroblasts ceils. The results of flow cytometry indicated that cell apoptosis increased after treated with Salvia mihiorrhiza injection when compared with TGF-β1 group （P 〈 0.05）. Meanwhile, Salvia miltiorrhiza injection could down regulate the expression of hydroxyproline （P 〈 0.01）. Conclusions Salvia miltiorrhiza injection can target human embryonic lung fibroblast cells, play a potent role in the airway remodeling through the promotion of its apoptosis and down regulate the expression of hydroxyproline.