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目的:探讨三七总皂苷(total saponins of panax notoginseng,PNS)对马兜铃酸I(aristolochic acid I,AAI)诱导的人肾小管上皮细胞株HK-2转分化的影响。方法:应用倒置相差显微镜观察HK-2细胞形态学变化;免疫组织化学方法检测α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达;ELISA法测定培养细胞上清液中转化生长因子-β1(transforming growth factor-β1,TGF-β1)的含量。结果:AA I诱导组HK-2细胞从原有典型的上皮细胞形态转变为长梭形肌成纤维细胞形态;胞浆内大量表达α-SMA,其积分光密度值增加(P<0.05);细胞培养上清液中TGF-β1含量均增加,且呈时间依赖性(P<0.05)。不同剂量PNS治疗可减轻AAI诱导的细胞形态学改变,不同程度的抑制α-SMA的表达和TGF-β1的分泌,且呈剂量依赖性(P<0.05)。PNS对照组HK-2细胞形态学、α-SMA表达和TGF-β1分泌与空白对照组比较均无明显变化。结论:AA I可通过促进TGF-β1的分泌诱导肾小管上皮细胞转分化,促进细胞外基质α-SMA的合成;PNS可抑制AA I诱导的肾小管上皮细胞转分化作用,减少α-SMA的表达,该作用可能是通过抑制TGF-β1表达实现的。
OBJECTIVE: To investigate the effects of total saponins of panax notoginseng (PNS) on the transdifferentiation of human renal tubular epithelial cell line HK-2 induced by aristolochic acid I (AAI). METHODS: The morphology of HK-2 cells was observed by inverted phase contrast microscopy. The expression of α-smooth muscle actin (α-SMA) was detected by immunohistochemical method. The changes in culture supernatants were detected by ELISA. The content of growth factor-β1 (TGF-β1). RESULTS: The HK-2 cells in the AA I-induced group changed from the original epithelial cell morphology to the long spindle-shaped myofibroblast cell morphology; a large amount of α-SMA was expressed in the cytoplasm and the integrated optical density value increased (P<0.05). The content of TGF-β1 in the cell culture supernatant increased, and it was time-dependent (P<0.05). Different doses of PNS treatment could reduce the AAI-induced cell morphological changes, inhibit the expression of α-SMA and TGF-β1 secretion to different extents in a dose-dependent manner (P<0.05). PNS control group HK-2 cell morphology, α-SMA expression and TGF-β1 secretion compared with the blank control group had no significant changes. Conclusion: AA I can induce the transdifferentiation of renal tubular epithelial cells by promoting the secretion of TGF-β1 and promote the synthesis of extracellular matrix α-SMA; PNS can inhibit the transdifferentiation of renal tubular epithelial cells induced by AA I and reduce the α-SMA level. Expression, this effect may be achieved by inhibiting the expression of TGF-β1.