ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2014
·Vol. 406
·期 28
·7379-7387
·
IF 3.578
Wang Jing,Liu Shu,Ma Bing,Chen Lina,Song Fengrui,Liu Zhiqiang,Liu Chun-ming
Xanthine oxidase (XOD) catalyzes the metabolism of hypoxanthine and xanthine to uric acid, the overproduction of which could cause hyperuricemia, a risk factor for gout. Inhibition of XOD is a major treatment for gout, and biflavonoids have been found to act as XOD-inhibitory compounds. In this study, ultrafiltration liquid chromatography with photodiode-array detection coupled to electrospray-ionization tandem mass spectrometry (UF-LC-PDA–ESI-MS) was used to screen and identify XOD inhibitors from S. tamariscina . High-performance counter-current chromatography (HPCCC) was used to separate and isolate the active constituents of these XOD inhibitors. Furthermore, ultrahigh-performance liquid chromatography (UPLC) and triple-quadrupole mass spectrometry (TQ-MS) was used to determine the XOD-inhibitory activity of the obtained XOD inhibitors, and enzyme kinetics was performed with Lineweaver–Burk (LB) plots using xanthine as the substrate. As a result, two compounds in S. tamariscina were screened as XOD inhibitors: 65.31 mg amentoflavone and 0.76 mg robustaflavone were isolated from approximately 2.5 g S. tamariscina by use of HPCCC. The purities of the two compounds obtained were over 98 % and 95 %, respectively, as determined by high-performance liquid chromatography (HPLC). Lineweaver–Burk plot analysis indicated that amentoflavone and robustaflavone were non-competitive inhibitors of XOD, and the IC
50 values of amentoflavone and robustaflavone for XOD inhibition were 16.26 μg mL −1 (30.22 μmol L −1 ) and 11.98 μg mL −1 (22.27 μmol L −1 ), respectively. The IC
50 value of allopurinol, used as the standard, was 7.49 μg mL −1 (46.23 μmol L −1 ). The results reveal that the method for systematic screening, identification, and isolation of bioactive components in S. tamariscina and for detecting their inhibitory activity using ultrafiltration LC–ESI-MS, HPCCC, and UPLC–TQ-MS is feasible and efficient, and could be expected to extend to screening and separation of other enzyme inhibitors.
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研究领域:
药理学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOMEDICINE & PHARMACOTHERAPY
·
2022
·
IF 7.419
Tianqiao Yong,Danling Liang,Chun Xiao,Longhua Huang,Shaodan Chen,Yizhen Xie,Xiong Gao,Qingping Wu,Huiping Hu,Xiangmin Li,Yuancao Liu,Manjun Cai
In this paper, we reported the hypouricemic effect of 2,4-dihydroxybenzoic acid methyl ester (DAE), a component of Ganoderma applanatum , in hyperuricemic mice through inhibiting XOD and down-regulating URAT1. Computationally, DAE showed a high similarity to allopurinol and depicted a high affinity in docking to XOD. In vitro , DAE exhibited an inhibitory effect against XOD. Importantly, DAE demonstrated a remarkable hypouricemic effect, decreasing serum uric acids (SUAs) of hyperuricemic mice (407 ± 31 μmol/L) to 195 ± 23, 145 ± 33 and 134 ± 16 μmol/L ( P < 0.01) at the doses of 20, 40, and 80 mg/kg with a dose-dependent manner and showing efficacies at 54–68 %, which were close to the efficacies of allopurinol (61 %) and benzbromarone (57 %). DAE depicted higher and negatively dose-independent urinary uric acids in comparison with that of the hyperuricemic control, implying DAE exerted an uricosuric effect and also a reduction effect on uric acid production. Unlike toxic allopurinol and benzbromarone, no general toxicity on body weights and no negative influence on liver, kidney, spleen and thymus were observed for DAE. Mechanistically, DAE inhibited XOD activities in vivo . Moreover, DAE up-regulated OAT1 and down-regulated GLUT9, URAT1 and CNT2. Overall, DAE may present a hypouricemic effect through inhibiting XOD and up-regulating OAT1 and down-regulating GLUT9, URAT1 and CNT2. This work provided novel insights into the hypouricemic effect of DAE and G. applanatum.
研究领域:
药理学
·
生物化学
·
分子生物学
·
医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Food Bioscience
·
2022
·
IF 5.318
Mengfan Hou,Huan Xiang,Xiao Hu,Shengjun Chen,Yanyan Wu,Jiachao Xu,Xianqing Yang
This study aimed to isolate and identify the novel potential Xanthine oxidase (XOD) inhibitory peptides derived from Trachinotus ovatus hydrolysate (TOH) (hydrolysis by neutral protease), and investigate their structure-function relationship. Initially, TOH was separated to four fractions (F1–F4) by gel filtration chromatography and F4 was chosen with higher XOD inhibitory rate (85.16% ± 0.566, 25 mg/mL) and lower molecular weight (95.73%, <1000 Da). F4 was further characterized by reversed-phase high performance liquid chromatography (RP-HPLC) coupled with Nano-HPLC-MS/MS and four peptides (FPAW, LLPW, WLLP and FHLP) were identified with IC 50 value of 3.81 ± 0.18 mM, 4.17 ± 0.12 mM, 43.06 ± 0.73 mM, and over 50.00 mM respectively. It can be concluded that Trp-containing peptides especially located in C-terminal could inhibit XOD effectively. Moreover, the XOD-inhibiting actions of these peptides involved π-π stacking (Phe914 and Phe1009 in XOD), hydrogen bonding via molecular docking . Lineweaver-Burk equation showed the inhibition type of FPAW was a mixed inhibition. Circular dichroism (CD) spectra indicated FPAW could combine and change the secondary structure of XOD. Therefore, two peptides of FPAW and LLPW might have potential to be used to alleviate hyperuricemia in the future.
研究领域:
生物化学
·
药理学
·
蛋白质组学
✓ 已验证
产品关联
DOI
高影响因子
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