This study aimed to investigate the effects of the total flavonoids of Radix Tetrastigma (RTF) on inflammation-related hepatocellular carcinoma (HCC) development. Extracted RTF was diluted to different concentrations for subsequent experiments. HCC cells were cotreated with lipopolysaccharide (LPS) and RTF to investigate the effects of RTF on LPS-stimulated HCC cells. A CCK-8 kit was used to measure cell proliferation. Apoptosis was detected with a flow cytometer. Cell migration and invasion were quantified by wound healing and Transwell assays, respectively. The expression of TLR4 and COX-2 and activation of the NF-κB pathway were determined by Western blotting. Treatment with LPS significantly enhanced cell proliferation and decreased the apoptosis rate, while cell migration and invasion were notably upregulated. RTF suppressed the proliferation and invasion induced by LPS stimulation and promoted HCC cell apoptosis. The protein levels of Bax and cleaved caspase-3 were decreased and that of Bcl-2 was increased by LPS in HCC cells, which could be rescued by RTF. RTF significantly inhibited the LPS-induced expression of the proinflammatory mediators IL-6 and IL-8 in HCC cells. Mechanistically, with RTF treatment, the upregulated expression of TLR4 and COX-2 induced by LPS was obviously downregulated. Furthermore, the phosphorylation of NF-κB/p65 was significantly decreased in LPS-stimulated cells after supplementation with RTF. Our study suggests that RTF exerts a significant inhibitory effect on the LPS-induced enhancement of the malignant behaviors of HCC cells via inactivation of TLR4/NF-κB signaling. RTF may be a promising chemotherapeutic agent to limit HCC development and inflammation-mediated metastasis.

这项研究旨在调查四味总黄酮（RTF）对炎症相关肝细胞癌（HCC）发育的影响。将提取的RTF稀释至不同浓度，以用于后续实验。将HCC细胞与脂多糖（LPS）和RTF共同处理，以研究RTF对LPS刺激的HCC细胞的影响。CCK-8试剂盒用于测量细胞增殖。用流式细胞仪检测细胞凋亡。细胞迁移和侵袭分别通过伤口愈合和Transwell测定来定量。通过Western印迹测定TLR4和COX-2的表达以及NF-κB途径的激活。LPS处理可显着增强细胞增殖并降低凋亡率，而细胞迁移和侵袭则显着上调。RTF抑制LPS刺激诱导的增殖和侵袭，并促进HCC细胞凋亡。LPS可以使HCC细胞中Bax和Caspase-3的蛋白水平降低，而Bcl-2的蛋白水平升高，这可以通过RTF挽救。RTF显着抑制LPS诱导的HCC细胞中促炎性介质IL-6和IL-8的表达。从机理上讲，通过RTF处理，LPS诱导的TLR4和COX-2的表达明显下调。此外，补充RTF后，LPS刺激的细胞中NF-κB/ p65的磷酸化明显降低。我们的研究表明，RTF通过灭活TLR4 /NF-κB信号传导，对LPS诱导的HCC细胞恶性行为增强具有显着的抑制作用。