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目的分析鲑鱼来源的聚脱氧核苷酸分子量分布对其生物学活性的影响。方法采用乙醇分级沉淀去纤苷类似物(defibrotide analogue,DFA),收集三步分级沉淀样品,脱水烘干,即H、M、L组分,体积排阻高效液相色谱(HPLC)法测定其分子量分布,S-2251底物法检测各分级组分对纤溶酶活性的影响,经典魏氏法检测各分级组分对红细胞沉降率的影响。结果获得H、M、L组分分别为0.46、1.03和0.38 g,80%H、M及L组分聚脱氧核苷酸链长分布分别为60~100、30~50及小于25 nt。DFA及H、M组分均可提高纤溶酶活性,与各组分分子量、浓度呈正相关,但L组分促纤溶活性不明显。与对照组相比,DFA及各分级组分均可引起红细胞沉降率显著升高,并呈剂量-效应关系,差异有统计学意义(P<0.05)。结论分子量分布是影响DFA活性的关键因素,为天然来源的聚脱氧核苷酸活性产物的筛选提供了参考。
Objective To analyze the effect of molecular weight distribution of salmon-derived poly-deoxynucleotides on their biological activity. Methods The defibrotide analogue (DFA) was precipitated by ethanol fractionation. The samples were fractionated and precipitated in three steps, dehydrated and dried, that is, H, M and L fractions were determined by size exclusion high performance liquid chromatography The molecular weight distribution and S-2251 substrate method were used to detect the effect of each fraction on the activity of plasmin. The effect of each fraction on the erythrocyte sedimentation rate was examined by classical Weiss method. Results The H, M and L fractions were 0.46, 1.03 and 0.38 g, respectively. The length distribution of polydeoxynucleotides of 80% H, M and L fractions were 60-100, 50-50 and less than 25 nt, respectively. DFA and H, M components can increase plasmin activity, and each component molecular weight, concentration was positively correlated, but L component fibrinolysis activity was not obvious. Compared with the control group, DFA and the graded components could cause the erythrocyte sedimentation rate to increase significantly, and there was a dose-effect relationship, the difference was statistically significant (P <0.05). Conclusion The molecular weight distribution is the key factor affecting the activity of DFA, which provides a reference for the screening of the active product of polydextrose nucleotide from natural sources.