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Anti-Helicobacter pylori activity of crude N-acetylneuraminic acid isolated from glycomacropeptide of whey

机译:从乳清糖巨肽中分离得到的粗制N-乙酰神经氨酸的幽门螺杆菌活性

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摘要

Helicobacter pylori colonizes the gastric mucosa of about half of the world's population, causing chronic gastritis and gastric cancer. An increasing emergence of antibiotic-resistant H. pylori arouses demand on alternative non-antibiotic-based therapies. In this study, we freshly prepared crude N-acetylneuraminic acid obtained from glycomacropeptide (G-NANA) of whey through a neuraminidase-mediated reaction and evaluated its antibacterial ability against H. pylori and H. felis. Overnight cultures of the H. pylori were diluted with fresh media and different concentrations (1-150 mg/mL) of crude G-NANA were added directly to the culture tube. Bacterial growth was evaluated by measuring the optical density of the culture medium and the number of viable bacteria was determined by a direct count of the colony forming units (CFU) on agar plates. For the in vivo study, mice were orally infected with 100 µL (5×108 cfu/mL) of H. felis four times at a day's interval, accompanied by a daily administration of crude G-NANA or vehicle. A day after the last infection, the mice were daily administered the crude G-NANA (0, 75, and 300 mg/mL) for 10 days and euthanized. Their stomachs were collected and bacterial colonization was determined by quantitative real-time PCR. Crude G-NANA inhibited H. pylori's growth and reduced the number of viable bacteria in a dose-dependent manner. Furthermore, crude G-NANA inhibited bacterial colonization in the mice. These results showed that crude G-NANA has antibacterial activity against Helicobacter and demonstrated its therapeutic potential for the prevention of chronic gastritis and gastric carcinogenesis induced by Helicobacter infection in humans.
机译:幽门螺杆菌定植在世界约一半人口的胃粘膜上,引起慢性胃炎和胃癌。耐药性幽门螺杆菌的出现越来越多,引起了人们对基于非抗生素的替代疗法的需求。在这项研究中,我们新鲜制备了通过神经氨酸酶介导的反应从乳清的糖巨肽(G-NANA)获得的粗制N-乙酰神经氨酸,并评估了其对幽门螺杆菌和猫屎肠杆菌的抗菌能力。用新鲜培养基稀释幽门螺杆菌的过夜培养物,并将不同浓度(1-150 mg / mL)的粗制G-NANA直接添加到培养管中。通过测量培养基的光密度来评估细菌的生长,并通过在琼脂平板上直接计数菌落形成单位(CFU)来确定活细菌的数量。在体内研究中,每天间隔四次对小鼠口服感染100 µL(5×10 8 cfu / mL)的猫屎肠球菌,并每天口服粗G-NANA或车辆。在最后一次感染后的第二天,每天向小鼠施用G-NANA粗品(0、75和300 mg / mL),持续10天并实施安乐死。收集他们的胃并通过定量实时PCR确定细菌定植。粗制的G-NANA以剂量依赖性方式抑制幽门螺杆菌的生长并减少活菌的数量。此外,粗G-NANA抑制了小鼠中的细菌定植。这些结果表明,粗制的G-NANA对幽门螺杆菌具有抗菌活性,并显示出其在预防人的幽门螺杆菌感染引起的慢性胃炎和胃癌发生中的治疗潜力。

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