沙棘乙酸乙酯萃取物化学成分分析及体外抑制幽门螺杆菌机理研究

Mechanism of Chemical components and anti-Helicobacter pylori sea buckthorn berries extract in vitro

沙棘是一种药食资源,在民间常应用于治疗胃部疾病。已有研究表明沙棘果乙酸乙酯萃取物具有较强的抗幽门螺杆菌活性。为探究沙棘乙酸乙酯萃取物抑制幽门螺杆菌的化学成分及抑制机理,试验采用高效液相色谱法测定其化学成分,并测定生物膜形成率、细胞膜通透性、膜蛋白荧光光谱、细胞膜完整性、脲酶活性和细胞微观结构等探究其抑菌机制。结果表明,沙棘乙酸乙酯萃取物中苹果酸含量最高,为872.97 mg/g,其次是芦丁、异鼠李素、槲皮素等黄酮类物质;沙棘乙酸乙酯萃取物可抑制生物膜形成,抑制率达到77.71%;细胞膜通透性增加2.45倍;膜蛋白结构发生变化,色氨酸和酪氨酸残基的亲水增强;脲酶活性下降87.89%;细胞内蛋白质和核酸的泄漏量达98.72%;扫描电镜结果表明萃取物会导致细胞结构变形,表面有褶皱。以上结果说明,沙棘乙酸乙酯萃取物中含有苹果酸和多种黄酮类物质,通过多种方式有效抑制幽门螺杆菌的生长繁殖,该结果为进一步分析沙棘中抑制幽门螺杆菌的活性物质奠定了基础。

Sea buckthorn is utilized to treat stomach disorders as a herb in China.It has been demonstrated that sea buckthorn ethyl acetate extract(EA)has exhibited an excellent ability to inhibit Helicobacter pylori.High-performance liquid chromatography was used to determine the chemical components of EA.Biofilm formation rate,cell membrane permeability,membrane protein fluorescence spectra,cell membrane integrity,urease activity,and cell microstructure were determined to investigate the mechanism of anti-H.pylori.Results showed that there was a high content of malic acid with 872.97 mg/g in EA,followed by rutin,isorhamnetin,quercetin,and other fla-vonoids.The EA inhibited biofilm formation with 77.71%,increased cell membrane permeability by 2.45 times,altered membrane pro-tein structure as tryptophan(Trp)and tyrosine(Tyr)residues to be more hydrophilic,and decreased the urease activity with 87.89%.The leakage rate of intracellular protein and nucleic acid reached 98.72%and EA distorted the structure and wrinkled the surface of the cell as the scanning electron microscope results.All the above indicated that EA contained malic acid and various flavonoids,which effec-tively inhibited the growth and reproduction of H.pylori in various ways.This result lays the foundation for further analysis of the active substances in sea buckthorn against H.pylori.