Abstract：
BACKGROUND: The aqueous extract of the dried buds of Syzygium aromaticum (SAAE) have the potential to alleviate Helicobacter pylori infection, but the specific molecular mechanism has not been fully elucidated.
OBJECTIVE: This study aimed to investigate the underlying mechanisms of SAAE on H. pylori pathogenicity.
METHODS: The inhibitory kinetics and anti-H. pylori adhesive capacity assays were conducted to examine the effects of SAAE on the growth and adhesive capability of H. pylori. The H. pylori outer membrane vesicles (OMVs) were purified from the culture supernatant through high-speed centrifugation, filtration, and two rounds of ultracentrifugation. Their characteristics and protein composition were then identified using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and qualitative proteomics study. Subsequently, the effect of SAAE on the pathogenicity of H. pylori OMVs was investigated using the Griess reagent assay, enzyme-linked immunosorbent assay (ELISA), quantitative proteomics study, TEM, and western blotting assay.
RESULTS: SAAE exhibited inhibitory effects on H. pylori growth and adhesion. The isolated H. pylori OMVs showed particle size of 27-242 nm and Zeta potential of -9.67 ± 0.53 mV. A total of 599 proteins were identified in the OMVs. Proteomics study indicated that the differential expressed proteins induced by OMVs with or without SAAE commonly enriched in P53 and autophagy pathways. Besides, SAAE counteracted the increased production of pro-inflammatory cytokines and attenuated the induction of cell autophagy caused by H. pylori OMVs. Furthermore, SAAE normalized the abnormal regulation of downstream targets (AIFM2 and IGFBP3) in the P53 signaling pathway caused by H. pylori OMVs.
CONCLUSIONS: SAAE can inhibit the growth and adhesion of H. pylori, reduce the inflammation and autophagy induced by H. pylori OMVs, and combated the abnormal regulation of P53 signaling pathway caused by H. pylori OMVs. These findings may help elucidate the mechanisms through which SAAE reduces the pathogenicity of H. pylori.
OBJECTIVE: This study aimed to investigate the underlying mechanisms of SAAE on H. pylori pathogenicity.
METHODS: The inhibitory kinetics and anti-H. pylori adhesive capacity assays were conducted to examine the effects of SAAE on the growth and adhesive capability of H. pylori. The H. pylori outer membrane vesicles (OMVs) were purified from the culture supernatant through high-speed centrifugation, filtration, and two rounds of ultracentrifugation. Their characteristics and protein composition were then identified using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and qualitative proteomics study. Subsequently, the effect of SAAE on the pathogenicity of H. pylori OMVs was investigated using the Griess reagent assay, enzyme-linked immunosorbent assay (ELISA), quantitative proteomics study, TEM, and western blotting assay.
RESULTS: SAAE exhibited inhibitory effects on H. pylori growth and adhesion. The isolated H. pylori OMVs showed particle size of 27-242 nm and Zeta potential of -9.67 ± 0.53 mV. A total of 599 proteins were identified in the OMVs. Proteomics study indicated that the differential expressed proteins induced by OMVs with or without SAAE commonly enriched in P53 and autophagy pathways. Besides, SAAE counteracted the increased production of pro-inflammatory cytokines and attenuated the induction of cell autophagy caused by H. pylori OMVs. Furthermore, SAAE normalized the abnormal regulation of downstream targets (AIFM2 and IGFBP3) in the P53 signaling pathway caused by H. pylori OMVs.
CONCLUSIONS: SAAE can inhibit the growth and adhesion of H. pylori, reduce the inflammation and autophagy induced by H. pylori OMVs, and combated the abnormal regulation of P53 signaling pathway caused by H. pylori OMVs. These findings may help elucidate the mechanisms through which SAAE reduces the pathogenicity of H. pylori.
摘要：
背景：香菇（SAAE）干燥芽的水提取物具有缓解幽门螺杆菌感染的潜力，但具体的分子机制尚未完全阐明。
目的：本研究旨在探讨SAAE对幽门螺杆菌致病性的潜在机制。
方法：抑制动力学和抗H。进行幽门螺杆菌粘附能力测定以检查SAAE对幽门螺杆菌生长和粘附能力的影响。通过高速离心从培养上清液中纯化幽门螺杆菌外膜囊泡(OMVs)，过滤,和两轮超速离心。然后使用透射电子显微镜（TEM）鉴定它们的特征和蛋白质组成，纳米粒子跟踪分析(NTA)，和定性蛋白质组学研究。随后，SAAE对H.pyloriOMV致病性的影响使用Griess试剂分析进行了研究，酶联免疫吸附测定(ELISA),定量蛋白质组学研究，TEM,和蛋白质印迹分析。
结果：SAAE对幽门螺杆菌的生长和粘附具有抑制作用。分离的幽门螺杆菌OMV显示27-242nm的粒径和-9.67±0.53mV的ζ电位。在OMV中鉴定了总共599种蛋白质。蛋白质组学研究表明，有或没有SAAE的OMV诱导的差异表达蛋白通常富含P53和自噬途径。此外,SAAE抵消了促炎细胞因子产生的增加，并减弱了幽门螺杆菌OMV引起的细胞自噬的诱导。此外，SAAE使幽门螺杆菌OMV引起的P53信号通路下游靶标(AIFM2和IGFBP3)的异常调节正常化。
结论：SAAE可以抑制幽门螺杆菌的生长和粘附，减少幽门螺杆菌OMV诱导的炎症和自噬，并对抗幽门螺杆菌OMV引起的P53信号通路的异常调节。这些发现可能有助于阐明SAAE降低幽门螺杆菌致病性的机制。
目的：本研究旨在探讨SAAE对幽门螺杆菌致病性的潜在机制。
方法：抑制动力学和抗H。进行幽门螺杆菌粘附能力测定以检查SAAE对幽门螺杆菌生长和粘附能力的影响。通过高速离心从培养上清液中纯化幽门螺杆菌外膜囊泡(OMVs)，过滤,和两轮超速离心。然后使用透射电子显微镜（TEM）鉴定它们的特征和蛋白质组成，纳米粒子跟踪分析(NTA)，和定性蛋白质组学研究。随后，SAAE对H.pyloriOMV致病性的影响使用Griess试剂分析进行了研究，酶联免疫吸附测定(ELISA),定量蛋白质组学研究，TEM,和蛋白质印迹分析。
结果：SAAE对幽门螺杆菌的生长和粘附具有抑制作用。分离的幽门螺杆菌OMV显示27-242nm的粒径和-9.67±0.53mV的ζ电位。在OMV中鉴定了总共599种蛋白质。蛋白质组学研究表明，有或没有SAAE的OMV诱导的差异表达蛋白通常富含P53和自噬途径。此外,SAAE抵消了促炎细胞因子产生的增加，并减弱了幽门螺杆菌OMV引起的细胞自噬的诱导。此外，SAAE使幽门螺杆菌OMV引起的P53信号通路下游靶标(AIFM2和IGFBP3)的异常调节正常化。
结论：SAAE可以抑制幽门螺杆菌的生长和粘附，减少幽门螺杆菌OMV诱导的炎症和自噬，并对抗幽门螺杆菌OMV引起的P53信号通路的异常调节。这些发现可能有助于阐明SAAE降低幽门螺杆菌致病性的机制。
