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该研究探讨了三七总皂苷对TGF-β1诱导的A549人肺泡上皮细胞间质转化和细胞外基质分解能力的影响。首先应用MTT法检测三七总皂苷对A549细胞增殖的影响,在12.5~200 mg·L~(-1),未发现三七总皂苷对A549细胞增殖有显著性的影响。接着将A549细胞分组。除正常组外,其余各组细胞应用5μg·L-1TGF-β1刺激,其中TGF-β1组正常培养,其他刺激组分别用50,100,200 mg·L~(-1)三七总皂苷干预。收集细胞和上清,先后采用圆形度值评估细胞的形态变化,免疫组化法检测特异性蛋白的表达以及酶联免疫吸附法检测MMP-9和TIMP-1的水平。TGF-β1刺激后,与正常组比较,细胞的形态发生变化,圆形度值显著减少(P<0.01);上皮特征性蛋白E-cad表达显著减少(P<0.05)而间质特征性蛋白α-SMA和FN的表达显著增加(P<0.05)。与TGF-β1组比较,三七总皂苷干预能显著增加圆形度值、降低α-SMA和FN的表达(P<0.05,P<0.01),但对E-cad的表达没有明显的影响。另外,TGF-β1刺激后,细胞分泌MMP-9的能力显著增强(P<0.05),而TIMP-1的分泌量未见显著变化。与TGF-β1组比较,三七总皂苷增加MMP-9的分泌量(P<0.05),减少TIMP-1的分泌量(P<0.05,P<0.01)。上述结果表明三七总皂苷能抑制肺泡上皮细胞的间质转化,促进细胞外基质分解,具有治疗肺纤维化的应用前景。
The study explored the Panax notoginseng on TGF-β1-induced A549 human alveolar epithelial cell stromal transformation and extracellular matrix degradation. At first, the effect of Panax notoginseng saponins on the proliferation of A549 cells was detected by MTT assay. No significant effect of Panax notoginseng saponins on the proliferation of A549 cells was found at 12.5 ~ 200 mg · L -1. A549 cells are then grouped. Except for the normal group, the other groups of cells were stimulated with 5μg · L-1 TGF-β1, TGF-β1 group of normal culture, the other stimulation group were 50,100,200 mg · L -1 Panax notoginseng intervention. The cells and supernatants were harvested and the morphological changes of the cells were evaluated by roundness values. The expression of specific proteins was detected by immunohistochemistry and the levels of MMP-9 and TIMP-1 by enzyme-linked immunosorbent assay. After TGF-β1 stimulation, the morphological changes of cells were observed and the degree of circularity was significantly decreased compared with the normal group (P <0.01). The expression of E-cadherin in epithelial cells was significantly decreased (P <0.05) The expression of α-SMA and FN increased significantly (P <0.05). Compared with the TGF-β1 group, the intervention of Panax notoginseng saponins could significantly increase the circularity, decrease the expression of α-SMA and FN (P <0.05, P <0.01), but had no obvious effect on the expression of E-cadherin. In addition, the ability of cells to secrete MMP-9 was significantly enhanced (P <0.05) after stimulated by TGF-β1, while the secretion of TIMP-1 did not change significantly. Compared with TGF-β1 group, Panax notoginseng saponin increased the secretion of MMP-9 (P <0.05) and decreased the secretion of TIMP-1 (P <0.05, P <0.01). The above results show that Panax notoginseng saponins can inhibit the mesenchymal transition of alveolar epithelial cells and promote extracellular matrix decomposition, with the application prospects of the treatment of pulmonary fibrosis.