非酒精性脂肪肝大鼠血清葡糖胺聚糖的二糖组成及皂苷对其影响

THE COMPOSITION OF THE DISACCHARIDES OF THE GLYCOSAMINOGLYCANS IN THE SERUM OF RATS WITH NONALCOHOL FATTY LIVER DISEASE AND EFFECTS OF SAPONIN

目的分析皂苷对乳清酸诱导的非酒精性脂肪肝模型(NAFLD)大鼠血清糖胺聚糖含量及硫酸软骨素/透明质酸(CS/HA)二糖组成的影响,筛选大鼠肝病的生物标志物。方法对NAFLD模型组大鼠分别喂食海参皂苷(Scc)、脱硫皂苷(Ds)、皂苷苷元(Ag)及海星皂苷(Sfc),离心层析分离纯化血清糖胺聚糖,硫酸咔唑法测定糖胺聚糖含量。糖胺聚糖酶解后2-氨基吖啶酮(AMAC)衍生,反相液相色谱检测二糖组成及相对含量。结果六组血清样本中,正常组总糖胺聚糖含量最低,NAFLD组、海参皂苷组、脱硫皂苷组、皂苷苷元组和海星皂苷组血清总糖胺聚糖含量分别增加了18.4%、23.2%、16.8%、21.0%、21.5%。分析二糖组成,0SCS在正常组最高(60.8%),模型组仅为21.0%(P<0.001)。四组皂苷组0SCS高于模型组,且皂苷苷元组最高(52.7%)。4SCS在对照组中最低(28%),模型组为68%(P<0.001)。结论 0SCS和4SCS可能是NAFLD标志物。海参皂苷(Scc)、脱硫皂苷(Ds)、皂苷苷元(Ag)及海星皂苷(Sfc)可治疗脂肪肝。

Objective To analyze the effect of saponin on the glyeosaminoglycan content and its chondroitin sulfate/ hyaluronic acid （CS/HA） disaccharides composition in the serum of orotic acid induced nonalcohol fatty liver disease （NAFLD） rats and screen the markers for liver diseases. Methods NAFLD rats were fed sea cucumber saponin, desulfurization saponin, saponin aglyeones, starfish saponins, or control diet, respectively. Serum glycosaminoglycans were separated and purified by Vivapure ion exchange centrifugal chromatography columns, and glycosaminoglycan content was measured using earbazole method. 2-aminoacridinone （AMAC） was derived after enzymolysis,and the disaceharides composition and relative content of glycosaminoglycan was analyzed by reverse-phase liquid chromatography （AMAC-HPLC） derivatization was used to analyze disaccharides composition and relative content of glycosaminoglycan after enzymolysis. Results For serum total glycosaminoglycan content, the control group was the lowest. However, Compareel with serum total glycosaminoglyean content of NAFLD group, sea cucumber saponin group, desulfurization saponin group, saponin aglycone group and starfish saponins group increased by 18.4%, 23.2%, 16.8%, 21%, 21.5%, respectively. For composition of disaccharides of glycosaminoglycan （GAG）, the relative content of GlcA-GalNAc （0SCS） in the control group （60.8%） was the highest, while it was only 21% （P〈0.001） in the model group. To compare the four saponins supplemented groups with the model group, the relative content of 0SCS was higher than that of the model group, and the relative content of 0SCS of the saponin aglycone group was 52.7% higher than the model group, The relative content of 4SCS of the control group was the lowest （28%）, whereas the model group was 68% （P〈0.001）. Conclusion 0SCS and 4SCS may be the markers for NAFLD. Sea cucumber saponin（Scc）, desulfurization saponin （Ds）,saponin aglycones （Ag） and starfish