Objective: This study is mainly to elucidate the characteristic components, hepatoprotective effect, and the underlying anti-hepatic fibrosis molecular mechanism of the total flavonoids of Tetrastigma hemsleyanum Diels & Gilg (SYQTF). Methods: The components of SYQTF were confirmed based on global natural products social molecular networking (GNPS) as well as the hepatoprotective effect on CCl4-induced acute liver injury. Cell toxicity and viability, cell cycle, and apoptosis analyses were then performed to determine the effects of SYQTF in transforming growth factor-β1 (TGF-β1) (10 ng/mL)-induced HSC-T6 hepatic fibrosis cell model. Finally, the lentiviral particles to package recombinant Ras-related C3 botulinum toxin substrate 1 (Rac1) over-expression and Rac1 knockdown plasmids were transfected into TGF-β1-induced HSC-T6 cells to elucidate the mechanism of anti-hepatic fibrosis of SYQTF targeting Rac1, respectively. Results: Our findings demonstrated that 86 components in SYQTF were identified in MSE and FastDDA mode with the aid of GNPS. Moreover, S15-ZJ, the genuine functional food and medicinal materials of T. hemsleyanum, was significantly different from other regions by multivariate statistical methods based on progenesis QI. Remarkably, kaempferol-3-O-furananose7-O-rhamnosyl-glucoside, the highest response intensity in S15-ZJ, may act as the potential Q-marker of genuine functional food and medicinal materials of T. hemsleyanum in Zhejiang Province distinguished from other areas. In addition, the in vivo experiments revealed that SYQTF greatly alleviated CCl4-induced acute liver injury. Furthermore, SYQTF can reversible the activation and proliferation of HSC-T6 cells, block the cell cycle in the G0/G1 phase, and promote cell apoptosis. Remarkably, the study is the first to reveal that SYQTF exhibited the anti-hepatic fibrosis effect through targeting Rac1 in the hedgehog signaling pathway. Conclusion: Rac1 is an important signaling molecule for the activation of the hedgehog pathway in HSC-T6. Importantly, SYQTF exerted its anti-hepatic fibrosis effect by inhibiting the Rac1-hedgehog signaling pathway. Furthermore, kaempferol-3-O-furananose7-O-rhamnosyl-glucoside could be a potential Q-marker of T. hemsleyanum in Zhejiang Province distinguished from other areas.
关键词:
抗肝纤维化;刺猬信号通路;Ras相关C3肉毒素底物1;三叶青;总黄酮
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Project Supported:
This research was supported by grants from the National Natural Science Foundation of China (No. 81803840), Zhejiang Traditional Chinese Medicine Scientific Research Program (No. 2021ZB083), and Zhejiang Cancer Hospital National Science Cultivation Foundation (No. PY2021020).
Xin?Wu?a?, Cuiwei?Chen?b?, Hongyan?Zhang?b, Keda?Lu?c *, Gang?Cao?a *. Exploration of chemical components and Rac1-dependent anti-hepatic fibrosis mechanism by total flavonoids derived from Tetrastigma hemsleyanum[J]. Chinese Herbal Medicines (CHM),2025,17(4):837-849
