背景:LamiophlomisHerba(LH)是在青藏高原发现的一种有价值的传统药用植物,可以促进血液循环,消除血瘀,并具有抗菌和抗炎特性。LH的主要成分是环烯醚萜苷,苯乙醇糖苷,黄酮类化合物,和多糖。
目的:探讨LH抗肝纤维化作用的机制并筛选其生物活性化合物。
方法:筛选LH标志物成分并验证LH抗肝纤维化机制。
方法:使用UPLC-Q-TOF-MS鉴定LH的活性成分,使用HotMap结合主成分分析(PCA)筛选标记成分。网络药理学和分子对接技术用于预测LH的潜在抗纤维化靶点。免疫荧光,酶联免疫吸附测定(ELISA),实时PCR(RT-PCR),和蛋白质印迹用于实验验证和机理研究。
结果:对该簇有积极贡献的15种化合物被鉴定为标记化合物。Acteoside,8-O-乙酰基山己苷甲酯(8-O-ASME),木犀草素,山芝甙甲酯(SME),Loganin,对数酸盐是主要的活性成分。网络药理学和分子对接研究表明,LH可能改善肝纤维化,炎症,和氧化应激,这可能与关键目标有关,如PTGS2,MAPK,EGFR,AKT1、SRC、Fn1、Col3a1、Col1a1和PC-III。ELISA的结果,RT-PCR和Westernblot实验表明,8-O-ASME,木犀草素,中小企业,Loganin,Loganate,LH组可以降低纤维连接蛋白的水平,Col1a1,Col3a1,α-SMA,Col-Ⅳ,LN,和PC-Ⅲ.
结论:LH改善HSC-T6细胞诱导的肝纤维化,并抑制细胞外基质(ECM)在肝细胞中的沉积,导致肝纤维化程度降低,具有良好的抗肝纤维化作用。
BACKGROUND: Lamiophlomis Herba (LH) is a valuable traditional medicinal plant found on the Qinghai-Tibetan Plateau that promotes blood circulation, removes blood stasis, and has antibacterial and anti-inflammatory properties. The main components of LH are iridoid glycosides, phenethyl alcohol glycosides, flavonoids, and polysaccharides.
OBJECTIVE: To investigate the mechanism of the anti-liver fibrosis effects of LH and screen for its bioactive compounds.
METHODS: Screening LH marker components and validating the LH anti-liver fibrosis mechanism.
METHODS: The active ingredients of LH were identified using UPLC-Q-TOF-MS, and HotMap combined with principal components analysis (PCA) was used to screen for marker components. Network pharmacology and molecular docking techniques were used to predict the potential anti-fibrotic targets of LH. Immunofluorescence, enzyme-linked immunosorbent assay (ELISA), real-time PCR (RT-PCR), and western blotting were used for experimental validation and mechanistic studies.
RESULTS: Fifteen compounds that actively contributed to the cluster were identified as marker compounds. Acteoside, 8-O-acetyl shanzhiside methyl ester (8-O-ASME), Luteolin, Shanzhiside Methyl ester (SME), Loganin, Loganate were the main active components. Network pharmacology and molecular docking studies have shown that LH might improve liver fibrosis, inflammation, and oxidative stress, which might be related to key targets such as PTGS2, MAPK, EGFR, AKT1, SRC, Fn1, Col3a1, Col1a1, and PC-III. The results of ELISA, RT-PCR and western blot experiments showed that Acteoside, 8-O-ASME, Luteolin, SME, Loganin, Loganate, and the LH group could reduce the levels of fibronectin, Col1a1, Col3a1, α-SMA, Col-Ⅳ, LN, and PC-Ⅲ.
CONCLUSIONS: LH improves liver fibrosis induced by HSC-T6 cells and inhibits the deposition of extracellular matrix (ECM) in hepatocytes, resulting in a decrease in the degree of liver fibrosis and a good anti-liver fibrosis effect.