PHYTOMEDICINE
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2025
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IF 8.3
Yufeng Deng,Qi hu,Zhiying Song,Mengyuan Nie,Zhiguo Wu,Yongrong Lei,Meng Liu,Dujuan Zhan,Baogang Xie
Objective Hepatocellular carcinoma (HCC) remains a leading cause of cancer mortality. Targeted therapies like lenvatinib face limitations due to resistance and immunosuppression. d -Glucaro-1,4-lactone (1,4-GL), a bioactive natural compound with anti-HCC activity, holds unexplored synergistic potential with lenvatinib. This study investigates the combinatorial efficacy of 1,4-GL and lenvatinib against HCC and elucidates the underlying mechanisms. Methods Efficacy was evaluated in H22 tumor-bearing mice treated with lenvatinib (20 mg/kg/day) ± 1,4-GL (25, 50, 100 mg/kg/day). Tumor inhibition, serum cytokines (TNF-α, IFN-γ, AFP), oxidative stress markers (MDA, SOD), liver enzymes (ALT, AST), and splenic T-cell subsets (flow cytometry) were assessed. Histopathology was analyzed via H&E staining. Lenvatinib pharmacokinetics (serum, liver, tumor) with/without 1,4-GL was determined in mice and SD rats using UPLC-MS/MS. In vitro, combinatorial effects on proliferation (CCK-8), migration (scratch assay), and clonogenicity (colony formation) were tested in Huh7 and HepG2 cells. STAT3 phosphorylation and PD-L1 expression (protein: Western blot; mRNA: RT-qPCR) were analyzed. PD-L1 induction was achieved using IFN-γ (20 ng/ml). Results The medium-dose combination (LMGL) achieved 89.97% tumor inhibition in H22 mice, significantly reducing tumor mass by 75% vs. lenvatinib alone (0.21±0.18 g vs. 0.84±0.79 g, p < 0.01). H&E staining revealed extensive necrosis in combination groups. Serum TNF-α and IFN-γ levels significantly increased ( p < 0.001), while AFP approached baseline. Flow cytometry showed an elevated CD4⁺/CD8⁺ T-cell ratio. Oxidative stress was modulated (reduced MDA and elevated SOD, p < 0.01). Pharmacokinetic studies revealed no significant differences in lenvatinib exposure or steady-state concentrations ( p > 0.05) with 1,4-GL co-administration. In vitro, 1,4-GL significantly reduced lenvatinib's IC 50 (Huh7: 18.54 μmol/l; HepG2: 30.34 μmol/l). Combinatorial treatment markedly downregulated PD-L1 protein/mRNA and inhibited STAT3 phosphorylation ( p < 0.05). Scratch and colony formation assays indicated synergy stemmed from immunomodulation and signaling inhibition, not enhanced anti-migration/proliferation. Conclusion 1,4-GL potently synergizes with lenvatinib against HCC by enhancing oxidative stress, reconstituting antitumor immunity, and suppressing the STAT3/PD-L1 signaling axis, without perturbing lenvatinib pharmacokinetics. This defines a novel phytochemical-based combinatorial strategy with significant clinical translation potential for HCC therapy.
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高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
·
2024
·
IF 4.8
Hongxin Cai,Yu Zhang,Jingyu Wang,Yufeng Deng,Jiangyuan Liu,Zhiguo Wu,Dejian Cao,Zhiying Song,Lele Wang,Baogang Xie
Ethnopharmacological relevance Liuwei dihuang pills is a famous Traditional Chinese Medicine with various anti-cancer properties. Over 50 pharmaceutical manufacturers produce Liuwei dihuang pills in China and an estimated millions of people around the world orally take it every day. D-glucaro-1,4-lactone (1,4-GL) was quantified to be about 12.0 mg/g in Liuwei dihuang pills and a primary bioactive component of it inhibiting the activity of β-glucuronidase in vivo. 1,4-GL can prevent and effectively inhibit various types of cancer. However, its exact mechanism of action remains unknown. The study would justify the traditional usage of Liuwei dihuang pills against cancers. Aim of the study 1,4-GL, a bioactive ingredient derived from Liuwei dihuang pills, a famous Traditional Chinese Medicine, could delay the progression of diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC) in rats. The mechanism underpinning the effect, however, remains poorly understood. Materials and methods Healthy and HCC rats were treated with or without 1,4-GL (40.0 mg/kg) and 1 HNMR-based metabonomic analysis was employed. 10 metabolites in uric acid pathway were quantitatively determined by UPLC-MS/MS. The expression of xanthine dehydrogenase (XDH), SLC2A9 mRNA, and SLC2A9 protein was determined using RT-qPCR and Western Blot. The effect of 1,4-GL on HCC-LM3 cells was verified in vitro. The alterations of ROS activity, SLC2A9 and XDH gene levels were observed in NCTC-1469 cells induced by lipopolysaccharide (LPS) after 1,4-GL treatment. Results After the intervention of 1,4-GL, improved pathological morphology, liver lesions in HCC rats was observed with restored serum levels of AFP, AST, ALP, γ-GGT and Fisher's ratio. Hepatic metabonomics revealed that puring metabolism were significantly regulated by 1,4-GL in HCC rats. Uric acid, xanthine and hypoxanthine levels were quantified by UPLC-MS/MS and found to be nearly restored to control levels after 1,4-GL treatment in HCC rats. Changes in xanthine oxidase activity, XDH mRNA expression, and SLC2A9 mRNA and protein expression were also reversed. 1,4-GL treatment in LM3 HCC cells were consistent with the results in vivo. Furthermore, oxidative stress indicators such as T-SOD, GSH, CAT and MDA in serum and liver were improved after HCC rats treated with 1,4-GL. In vitro, 1,4-GL was observed to reduce lipopolysaccharide-induced ROS levels in NCTC-1469 cells with enhanced mRNA and protein expression of SLC2A9 and decreased mRNA level of XDH. Conclusion The protective effects of 1,4-GL against DEN-induced HCC by reducing uric acid and ROS levels due to down-regulation of uric acid production and up-regulation of SLC2A9 expressions. 1,4-GL may represent a novel treatment that improves recovery from HCC by targeting uric acid-ROS pathway.
研究领域:
药理学
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肿瘤学
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传统中药
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分子生物学
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生物化学
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高影响因子
Analytica
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2022
·Vol. 3
·期 1
·36-53
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IF 0
Andrey A. Chudin,Elena V. Kudryashova
Reversed micelles are helpful to solubilize otherwise insoluble membranotropic or membrane-bound enzymes in their functional form, thus enabling activity assay and inhibition analysis. However, in the case of redox enzymes, this task is further complicated by the necessity to select an appropriate electron-acceptor (EA) which, ideally, should be compatible with spectrophotometric measurements in reversed micelles. Here, we have identified such an EA and successfully used it in a reversed micellar environment to assay the activity of two homologous enzymes from mitochondria:l-galactone-1,4-lactone dehydrogenase (EC 1.3.2.3) fromArabidopsis thaliana(AtGALDH) and galactonolactone oxidase (EC 1.3.3.12) fromTrypanosoma cruzi(TcGAL), differing in their membranotropic properties, with TcGAL being almost insoluble in water and particularly difficult to assay. Furthermore, we have demonstrated the possibility to use this assay for inhibition analysis, with an elucidation of the mechanism and inhibition parameters, which otherwise could not be possible. In order to perform inhibition analysis, we improved the approach for the determination of activity of such membrane enzymes based on a reversed micellar system as membrane matrix, necessary for the functioning of membrane enzymes. A number of electron acceptors (EA) were tested for AtGALDH and optimal conditions of activity determination for AtGALDH were found. The suggested method was successfully applied to the study of the inhibition of AtGALDH by lycorine, and the mixed competitive mechanism of inhibition of AtGALDH by lycorine was determined. The developed approach to inhibitor analysis was applied for TcGAL, insoluble in water membrane, and the method provides new opportunities for searching effective inhibitors that may be potential drugs. Indeed, galactonolactone oxidase fromTrypanosoma cruzi(TcGAL) and AtGALDH are homologues, and the inhibition of TcGAL stops the vital biosynthesis of vitamin C in parasiteTrypanosoma cruzifrom causing Chagas disease. The approach proposed can be applied for the screening of inhibitors of AtGALDH and TcGAL, as well as to study properties of other membrane enzymes including determination of the mechanisms of inhibition, structure and catalytic properties, the impact of membrane components (for example lipids), and so on.Keywords:determination of membrane enzyme activity;spectrophotometric method;membrane enzymes;l-galactone-1,4-lactone dehydrogenase;Arabidopsis thaliana;galactonolactone oxidase;Trypanosoma cruzi;lycorine;drugs;Chagas disease
研究领域:
生物学
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生物化学
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药理学
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寄生虫学
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Probiotics and Antimicrobial Proteins
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2025
·
IF 4.4
Xu Yazhou,Xiong Tao,Zhang Linli,Kang Xinyi,Ma Fang,Zhao Mingwei
Type 2 diabetes (T2D) and its associated complications have emerged as significant global public health challenges. Postbiotics have shown potential benefits in managing T2D. To identify effective postbiotics for T2D amelioration and elucidate the material basis underlying their efficacy, this study developed an anti-hyperglycemic evaluation model. Using an LC–MS platform combined with bioinformatics approaches, we comprehensively analyzed the metabolic distribution profiles of postbiotics in cell-free supernatant (CFS) and cell-free extract (CFE). Through this analysis, we identified a series of notable differential metabolites, including alpha amino acids, D-gulono-1,4-lactone, and glutathione. Spectrum-effect analysis was employed to preliminarily investigate the functional active substances responsible for the anti-hyperglycemic effects of postbiotics, revealing compounds potentially associated with anti-hyperglycemic properties, such as pyroglutamic acid and fumaric acid. Additionally, ultra-filtration mass spectrometry experiments were conducted to target and isolate potential bioactive compounds interacting with α-amylase, α-glucosidase, and tyrosine phosphatase 1B (PTP-1B), including mesoxalic acid, γ-glutamylphenylalanine, and L-3-phenyllactic acid.
研究领域:
糖尿病研究
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微生物学
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药理学
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代谢组学
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生物信息学
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食品科学
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生物技术
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高影响因子
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