摘要
本研究旨在研究染料木黄酮(Genistein,Gen)对大鼠体内N-羟乙酰神经氨酸(N-glycolylneuraminic acid,Neu5Gc)生物合成的影响。选取80只4周龄SD雄性大鼠,随机平均分为对照组和Gen组,分别灌胃5%的乙醇溶液和300 mg/(kg·d)的Gen溶液。利用荧光高效液相色谱(HPLC-FLD)检测大鼠后腿肌肉、肾脏、肝脏组织中Neu5Gc的含量,并采用Gen与唾液酸转移酶(Sialyltransferase,ST)分子对接,初步探讨了其抑制Neu5Gc合成的机理。结果表明:灌胃15 d时,后腿肌肉和肝脏组织中的Neu5Gc的含量分别降低了13.77%和15.45%,而肾脏组织中Neu5Gc的含量变化差异不显著;30 d时,在肌肉组织中未检出Neu5Gc,在肝脏组织中的Neu5Gc的含量降低了13.35%,肾脏组织中Neu5Gc的含量没有显著的变化;45 d时,在后腿肌肉、肾脏组织和肝脏组织中的Neu5Gc含量分别降低了32.65%、16.80%和32.78%;60d时,在后腿肌肉、肾脏组织和肝脏组织中Neu5Gc含量降低了12.72%、12.30%和11.42%。Gen与ST活性位点残基His319、Ser151、Gly293、Thr328形成氢键,且与残基His302、His301、Trp300、Ser271、Phe292、Thr328、Ser325、Ile274形成疏水作用。因此分子间弱相互作用是导致Gen抑制ST活性的主要原因。该研究结果为后续开展宰前降低红肉中Neu5Gc的方法提供了基础实验方法支撑。
To investigate the effects of genistein(Gen)on the biosynthesis of N-glycolylneuraminic acid(Neu5 Gc)in rats,80 4-week-old male SD rats were randomly equally into the control and genistein groups.The rats of control and genistein groups were fed 5%ethanol and 300 mg/(kg·d)genistein respectively by gavage.The contents of Neu5 Gc in hind leg muscle,kidney and liver tissues of rats were measured by using high performance liquid chromatography coupled with fluorescence detector(HPLC/FLD),and the mechanism of inhibition of Neu5 Gc synthesis was investigated by using the molecular docking of Gen and sialyltransferase.On the 15 th day,the content of Neu5 Gc in hind leg muscle and liver tissues decreased 13.77%and 15.45%,respectively,and there was no significant change in the content of Neu5 Gc in kidney tissues.On the 30 th day,the content of Neu5 Gc in liver tissues decreased 13.35%,however,there was no significant change in the content of Neu5 Gc in kidney tissues and Neu5 Gc was not detected in hind leg muscle.The content of Neu5 Gc in hind leg muscle,kidney and liver tissues decreased respectively 32.65%,32.78%,16.80%and 12.72%,11.42%,12.30%while rats fed on the 45 th and the 60 th days.Genistein has formed the hydrogen bond with sialyltransferase activity site residues His319,Ser151,Gly293,Thr328 and formed a hydrophobic interactions with the residues His302,His301,Trp300,Ser271,Phe292,Thr328,Ser325 and Ile274.The results of molecular docking indicated that the weak intermolecular interaction was the main cause of genistein inhibiting sialyltransferase activity.The research results provided an experimental basis for the subsequent reduction of Neu5 Gc in red meat before slaughter.
作者
李洪英
常瑞
朱秋劲
朱旭玲
徐阿奇
周樱子
晏印雪
Hongying Li;Rui Chang;Qiujin Zhu;Xuling Zhu;Aqi Xu;Yingzi Zhou;Yinxue Yan(School of Liquor and Food Engineering,Guizhou University,Guiyang 550025,Guizhou,China;Guizhou Province Key Laboratory of Agricultural and Animal Products Storage and Processing,Guiyang 550025,Guizhou,China)
出处
《生物工程学报》
CAS
CSCD
北大核心
2019年第5期857-870,共14页
Chinese Journal of Biotechnology
基金
国家自然科学基金(No.31660496)
贵州省高层次创新型人才培养项目(黔科合平台人才No.[2016]5662)
贵州大学研究生创新基金(No.GDTSSPGZZ 201707)资助~~
