The effect of total saponins of Schizocapsa plantaginea Hance on epithelial-mesenchymal transition of rat hepatic oval cell line WB-F344
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摘要 目的：探讨裂果薯总皂苷（SSPHs）对转化生长因子-β1（TGF-β1）诱导的大鼠肝卵圆细胞系 WB-F344 上皮间质转化（EMT）的影响及作用机制。方法：将WB-F344细胞分为对照组、模型组，SSPHs低、中、高剂量组和PI3K/AKT信号通路抑制剂LY294002组。除对照组外，其余各组均以10 μg/L TGF-β1诱导WB-F344构建EMT模型。采用MTT法、细胞划痕实验分别检测细胞增殖、迁移能力，实时荧光定量PCR（RT-qPCR）、western blotting法分别检测E-钙黏蛋白（E-cadherin）、N-钙黏蛋白（Ncadherin）、波形蛋白（Vimentin）的mRNA和蛋白表达，western blotting法检测PI3K/AKT信号通路关键蛋白表达。结果：SSPHs可抑制TGF-β1诱导的WB-F344细胞的增殖，24 h的细胞半数抑制浓度（IC50）为3.02 μg/mL。与对照组比较，模型组N-cadherin、Vimentin的mRNA及蛋白表达水平显著升高，E-cadherin mRNA及蛋白表达水平显著降低，p-PI3K/PI3K、p-AKT/AKT比值显著升高（均 P＜0.05）。与模型组比较，SSPHs 中、高剂量组 N-cadherin 和 Vimentin mRNA 及蛋白表达下调，E-cadherin mRNA 及蛋白表达上调，p-PI3K/PI3K、p-AKT/AKT 比值降低（均 P＜0.05），结果与 LY294002 组相似。结论：SSPHs 可抑制TGF-β1诱导的WB-F344细胞EMT进程，其机制可能与抑制PI3K/AKT信号通路有关。Abstract Objective: To investigate the effect and mechanism of total saponins of Schizocapsa plantaginea Hance (SSPHs) on epithelial mesenchymal transformation (EMT) of rat hepatic oval cells WB-F344 induced by transforming growth factor- β1 (TGF- β1). Methods: WB- F344 cells were divided into control group, model group, low - , medium - and high- dose SSPHs groups and PI3K/AKT signaling pathway inhibitor LY294002 group. Except the control group, all the other groups were induced with 10 μg/L TGF-β1 to establish the EMT model. MTT assay and cell scratch assay were used to detect cell proliferation and migration. Reverse transcription-quantitative PCR (RT-qPCR) and western blotting were used to detect the mRNA and protein expression of E-cadherin, N-cadherin and Vimentin, respectively. The expression of key proteins in PI3K/AKT signaling pathway was detected by western blotting. Results: SSPHs inhibited the proliferation of TGF-β1-induced WB-F344 cells, with a half maximal inhibitory concentration (IC50) value of 3.02 μg/mL at 24 hours. Compared with the control group, the model group showed significantly increased mRNA and protein expression levels of N- cadherin and Vimentin, significantly decreased mRNA and protein expression of E- cadherin, and significantly increased ratio of p-PI3K/PI3K and p-AKT /AKT (all P＜0.05). Compared with the model group, N-cadherin and Vimentin mRNA and protein expression was down-regulated, E-cadherin mRNA and protein expression was upregulated, and P-PI3K/PI3K and P-AkT/AKT ratios were decreased in the THE SSPHs medium and high-dose groups (all P＜0.05), with results similar to those observed in the LY294002 group (all P＜0.05). Conclusion: SSPHs can inhibit the EMT process in TGF-β1-induced WB-F344 cells, and its mechanism may be related to the inhibition of PI3K/AKT signaling pathway.
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