Tang Ying, Yang Fan, Wen Xia, Zhou Yi, Tang Rong, He Xiuzhi, Lu Qiang, Li Cailan
Department of Pharmacology, Zunyi Medical University, Zhuhai, China.
Department of Pharmaceutical Sciences, Zunyi Medical University, Zhuhai, China.
Front Cell Infect Microbiol. 2025 Jul 25;15:1617330. doi: 10.3389/fcimb.2025.1617330. eCollection 2025.
Roxb. (SGR), known as "tufuling" in China, is a medical and edible plant, which has anti-inflammatory, antibacterial and antineoplastic activity. SGR is extensively utilized in the remedy of gastroenteric disorders associated with infection. However, the precise mechanism underlying the anti- function of SGR remains to be elucidated.
The inhibitory impact of SGR on the growth of was examined. Subsequently, SGR against urease (HPU) and jack bean urease (JBU) was investigated to illuminate the inhibitory effects, kinetic types, sites of inhibition, and potential mechanisms of action.
UPLC-ESI-MS/MS was applied to identify the components of SGR. The anti- effect of SGR was conducted by agar dilution method. The enzyme inhibitory activities of SGR and its primary constituents were assessed through a modified spectrophotometric Berthelot (phenol-hypochlorite) assay. The kinetics of urease inhibition were analyzed using Lineweaver-Burk plots. To explore the underlying mechanisms, sulfhydryl group reagents and Ni binding depressors were employed. Additionally, molecular docking simulations were conducted to examine the binding interactions between the main compounds of SGR and urease.
A total of 34 compounds including astilbin, engeletin, isoengeletin, neoastilbin, isoastilbin and neoisoastilbin are identified in SGR. SGR was observed to inhibit the growth of three strains (ATCC 43504, NCTC 26695, and ICDC 111001) with minimum inhibitory concentration (MIC) values spanning a range of 0.5 to 1.5 mg/mL. Moreover, SGR exerted a significant inhibitory effect on HPU and JBU, with IC values of 1.04 ± 0.01 mg/mL and 1.01 ± 0.01 mg/mL, separately. Enzyme kinetics analysis showed that SGR was a slow binding, non-competitive depressor to HPU, and a slow binding, mixed depressor to JBU. In-depth mechanism exploration showed that thiol compounds had better protective effect on HPU or JBU than inorganic substances, implying that the active site of SGR repressing urease may be the sulfhydryl group. Furthermore, glutathione reactivated SGR-inhibited urease, demonstrating that the inhibition was reversible. Additionally, astilbin and engeletin exhibited a certain inhibitory role towards urease activity, with astilbin inhibiting urease more than three times as strongly as engelitin. Enzyme kinetics analysis established that the inhibitory role of astilbin on enzymes was consistent with that of SGR. Molecular docking study indicated that astilbin and engeletin interacts with sulfhydryl groups at the active site of urease.
These results indicated that SGR could prominently inhibit growth through targeted suppression of its secreted urease. This investigation provides substantial experimental evidence supporting the consideration of SGR as a safe and promising natural treatment for -associated gastrointestinal diseases.
虎杖（SGR）在中国被称为“土茯苓”，是一种药食两用植物，具有抗炎、抗菌和抗肿瘤活性。SGR被广泛用于治疗与感染相关的胃肠道疾病。然而，SGR抗炎作用的确切机制仍有待阐明。
研究SGR对幽门螺杆菌生长的抑制作用。随后，研究SGR对幽门螺杆菌脲酶（HPU）和刀豆脲酶（JBU）的抑制作用，以阐明其抑制效果、动力学类型、抑制位点及潜在作用机制。
采用超高效液相色谱-电喷雾串联质谱（UPLC-ESI-MS/MS）鉴定SGR的成分。通过琼脂稀释法检测SGR的抗菌作用。采用改良的分光光度法贝托洛（酚-次氯酸盐）测定法评估SGR及其主要成分的酶抑制活性。使用Lineweaver-Burk图分析脲酶抑制动力学。为探索潜在机制，使用巯基试剂和镍结合抑制剂。此外，进行分子对接模拟以研究SGR主要化合物与脲酶之间的结合相互作用。
在SGR中鉴定出34种化合物，包括落新妇苷、柚皮素、异柚皮素、新落新妇苷、异落新妇苷和新异落新妇苷。观察到SGR可抑制三种幽门螺杆菌菌株（ATCC 43504、NCTC 26695和ICDC 111001）的生长，最低抑菌浓度（MIC）值范围为0.5至1.5 mg/mL。此外，SGR对HPU和JBU均有显著抑制作用，IC值分别为1.04±0.01 mg/mL和1.01±0.01 mg/mL。酶动力学分析表明，SGR对HPU是慢结合、非竞争性抑制剂，对JBU是慢结合、混合型抑制剂。深入的机制探索表明，硫醇化合物对HPU或JBU的保护作用优于无机物，这意味着SGR抑制脲酶的活性位点可能是巯基。此外，谷胱甘肽可使被SGR抑制的脲酶重新激活，表明该抑制作用是可逆的。此外，落新妇苷和柚皮素对脲酶活性具有一定的抑制作用，落新妇苷对脲酶的抑制作用比柚皮素强三倍以上。酶动力学分析表明，落新妇苷对酶的抑制作用与SGR一致。分子对接研究表明，落新妇苷和柚皮素与脲酶活性位点的巯基相互作用。
这些结果表明，SGR可通过靶向抑制幽门螺杆菌分泌的脲酶显著抑制其生长。本研究提供了大量实验证据，支持将SGR视为治疗幽门螺杆菌相关胃肠道疾病的一种安全且有前景的天然疗法。
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