肝素衍生物结构与抑制高糖引起人系膜细胞增殖的关系研究

Structure-activity relationship of heparin derivatives structures and their antiproliferation activity on high glucose-induced mesangial cell proliferation

目的考察肝素结构与抑制高糖引起人肾小球系膜细胞增殖活性之间的关系。方法采用化学修饰的方法制备2,3-O-脱硫酸肝素、6-O-脱硫酸肝素、N-脱硫酸肝素、高碘酸氧化/硼氢化钠还原肝素、羧基还原肝素和多硫酸化肝素,通过比浊法测得硫酸基团含量,核磁图谱分析肝素修饰物;MTT法比较这些肝素修饰物对高糖引起人肾小球系膜细胞增殖的抑制活性。结果多硫酸化肝素抑制活性显著增加,羧基还原肝素、高碘酸氧化/硼氢化钠还原肝素抑制活性相对肝素没有明显改变,而2,3-O-脱硫酸肝素、6-O-脱硫酸肝素和N-脱硫酸化肝素抑制活性显著降低。结论抑制活性大小的顺序和硫酸基团含量大小顺序一致,说明肝素中硫酸基团的含量是影响肝素抑制高糖引起人肾小球系膜细胞增殖的主要因素。

Purpose To investigate the relationship between the structure of heparin derivatives and antiproliferative activity on high glucose-induced mesangial cell proliferation. Methods Preparation of 2, 3-O- desulfated hcparin, 6- O-desulfated heparin, N-desulfated heparin, periodate-oxidized/borohydride-reduced heparin, carboxyl-reduced heparin and poly-sulfated heparin by chemical method. Sulfate content （SO4^2- , % ） was determined by turbidimetrical asbarium sulfate and the structure of chemically modified heparin was verified by NMR spectroscopy. The anti-proliferative activities were assayed by MTT. Results Compared to heparin, the anti-proliferative activity of poly-sulfated heparin was significantly increased and the activity of periodate-oxidized/borohydride-reduced heparin and carboxyl-reduced heparin had no obvious change, while the 2,3- O-desulfated,6- O-desulfated and N-desulfated heparin was significantly decreased. Conclusion The order of anti-proliferative activities of chemically modified heparin is consistent with the content of SO4^2- . These results indicated that the content of SO4^2- is the major factor of heparin＇s structure for the inhibitive effect on high glucose-induced mesangial cell proliferation.