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目的探讨薏苡仁油对人肺泡上皮细胞间质转化的影响,评估薏苡仁油在肺纤维化治疗方面的应用。方法以A549肺泡上皮细胞为研究对象,用TGF-β1诱导肺泡上皮细胞的间质转化,同时,加入不同浓度的薏苡仁油进行干预。TGF-β1刺激48 h后,应用圆形度定量分析细胞形态的变化;应用免疫细胞化学的方法分析细胞表面上皮特征性蛋白E-cadherin和间质特征性蛋白α-SMA、FN和Collagen I的表达;应用双抗体夹心酶联免疫吸附法检测细胞分泌MMP-9和TIMP-1水平。结果与正常组比较,TGF-β1组圆形度显著降低(P<0.01);E-cadherin表达显著降低(P<0.05);FN表达显著升高(P<0.01);α-SMA和Collagen I表达升高,MMP-9和TIMP-1的分泌量增加,但均无显著性(P>0.05)。与TGF-β1组比较,高剂量组E-cad表达显著增加(P<0.05);小剂量组Collagen I的表达显著降低(P<0.05);小剂量组TIMP-1分泌量显著增加(P<0.05),其它未显示出显著性的影响(P>0.05)。结论薏苡仁油能有效抑制体外培养的人肺泡上皮细胞A549的上皮间质转化,抑制MMPs对细胞外基质的破坏。
Objective To investigate the effect of Coix Seed Oil on the proliferation of human alveolar epithelial cells and to evaluate the application of Coix Seed Oil in the treatment of pulmonary fibrosis. Methods The A549 alveolar epithelial cells were used as the research object. The interstitial transformation of alveolar epithelial cells was induced by TGF-β1. At the same time, different concentrations of Coix seed oil were used for intervention. After 48h stimulation with TGF-β1, the morphological changes of cells were quantitatively analyzed by circularity. The expressions of E-cadherin, interstitial proteins α-SMA, FN and Collagen I were analyzed by immunocytochemistry The levels of MMP-9 and TIMP-1 secreted by cells were detected by double antibody sandwich enzyme-linked immunosorbent assay. Results Compared with the normal group, the degree of circularity of TGF-β1 group was significantly decreased (P <0.01), the expression of E-cadherin was significantly decreased (P <0.05), FN expression was significantly increased (P <0.01) The expression of MMP-9 and TIMP-1 increased, but there was no significant difference (P> 0.05). Compared with TGF-β1 group, the expression of E-cadherin in high-dose group was significantly increased (P <0.05); the expression of Collagen I in low-dose group was significantly decreased (P <0.05); TIMP- 0.05), others did not show significant effects (P> 0.05). Conclusion Coix seed oil can effectively inhibit the epithelial-mesenchymal transition of human alveolar epithelial A549 in vitro and inhibit the destruction of extracellular matrix.