摘要
目的优化扁藻胞外多糖的磷酸化工艺并研究磷酸化扁藻胞外多糖对肿瘤的抑制作用。方法以正交实验分析影响扁藻胞外多糖磷酸化的因素,分别检测扁藻胞外多糖(PEPs)和磷酸化扁藻胞外多糖(pPEPs)的红外吸光度,利用MTT法研究扁藻胞外多糖及其磷酸化衍生物对Raw 264.7的生长抑制作用。结果正交实验显示最佳多糖磷酸化工艺条件为:反应温度为100℃,反应时间为6 h,磷酸化试剂比例为6∶1,pH=8。PEPs与pPEPs样本经红外吸光度扫描显示多糖均为吡喃型多糖,以共价键形式连接的磷酸化多糖修饰成功。MTT结果显示PEPs与pPEPs均对小鼠巨噬细胞Raw 264.7的增殖具有抑制作用(P<0.05或0.01),且以pPEPs的抑制作用更为显著(P<0.05或0.01)。结论以正交实验统计方法筛选扁藻胞外多糖磷酸化工艺操作简单方便,pPEPs具有体外抗肿瘤作用。
Objective To optimize the phosphorylation process of platymonas exopolysaccharides(PEPs)and investigate the inhibitory effect of phosphorylated platymonas exopolysaccharides(pPEPs)on tumors.Methods The factors affecting the pPEPs were analyzed with orthogonal experiment.The infrared absorbance of the PEPs and pPEPs was detected respectively.The growth inhibition effect of the PEPs and pPEPs on Raw 264.7 cells was studied with the MTT method.Results Orthogonal experiments showed that the optimum polysaccharide phosphorylation process conditions were:reaction temperature of 100℃,reaction time of 6 h,phosphorylation reagent ratio of 6∶1,and pH=8.Infrared absorbance scanning indicated that the PEPs and pPEPs were all pyran polysaccharide,and phosphorylated polysaccharides linked by covalent bonds were successfully modified.MTT results showed that both PEPs and pPEPs had inhibitory effects on the proliferation of mouse macrophage Raw 264.7(P<0.05 or 0.01),and the inhibitory effect of pPEPs was more significant(P<0.05 or 0.01).Conclusion It is simple and convenient to screen the process of the pPEPs with orthogonal experimental.The pPEPs have anti-tumor effect in vitro.
作者
易志恒
翁正芬
温俊昊
刘梓茂
罗辉
吴斌华
YI Zhi-heng;WENG Zheng-fen;WEN Jun-hao;LIU Zi-mao;LUO Hui;WU Bin-hua(Institute of Marine Medicine,Guangdong Key Laboratory for Research and Development of Natural Drugs,and the First School of Clinical Medicine,Guangdong Medical University,Zhanjiang 524023,China;Southern Marine Science and Engineering Guangdong Laboratory(Zhanjiang),Zhanjiang 524023,China;Guangdong Institute of Marine Medicine,Zhanjiang 524023,China)
出处
《广东医科大学学报》
2022年第5期498-502,507,共6页
Journal of Guangdong Medical University
基金
广东省科技计划项目(2016B030309002)
广东医学院科研基金项目(2XK14053)。
关键词
扁藻
胞外多糖
磷酸化修饰
正交实验
抗肿瘤
platymonas
exopolysaccharides
phosphorylation
orthogonal experiment
anti-tumor
