目的 对比研究朝鲜淫羊藿酸性多糖酯化还原前后的理化特性，并探讨其改善油酸诱导的肝癌HepG2细胞脂质堆积活性的差异。方法 采用高效凝胶渗透色谱法测定朝鲜淫羊藿酸性多糖（EFPA）的均一性和分子量，高效液相色谱法测定EFPA和酯化还原后朝鲜淫羊藿酸性多糖（EFPA-R）的单糖组成；采用油酸（OA）处理HepG2细胞诱导建立脂质蓄积模型，不同浓度EFPA与EFPA-R（10、30、100、300 μg·mL^-1）分别和OA共同作用于细胞24 h，采用CCK-8试剂盒测定细胞存活率，油红O染色观察细胞内脂滴蓄积情况，并采用试剂盒测定细胞内总胆固醇（TC）、甘油三酯（TG）含量。结果 EFPA为成分均一的多糖组分，分子量为125.8 kDa，由甘露糖、葡萄糖、半乳糖、葡萄糖醛酸和阿拉伯糖组成，摩尔比为1.7∶7.4∶1.4∶1.8∶1.0，葡萄糖占比最大，EFPA-R由甘露糖、葡萄糖、半乳糖和阿拉伯糖组成，摩尔比为0.8∶10.6∶2.1∶1.0；在10-300 μg·mL^-1范围内，EFPA和EFPA-R对HepG2细胞的抑制作用较弱，作为给药浓度；与空白组相比，模型组细胞中TC、TG含量显著升高（P < 0.01），细胞内红色脂滴显著增多，与模型组相比，EFPA可显著降低细胞中TC、TG含量（P < 0.01），明显减少细胞内红色脂滴（P < 0.05或P < 0.01），EFPA-R干预后细胞则无明显变化。结论 EFPA可明显改善HepG2细胞脂质堆积情况，且呈现剂量依赖性，而半乳糖醛酸（GalA）的存在可能是其抑制HepG2细胞脂质蓄积的关键因素。

Objective To compare the structural characteristics and the difference on its effects on the reduction of lipogenesis in oleic acid-induced HepG2 cells of the acidic polysaccharide isolated from Epimedium koreanum Nakai.Methods The molecular weight and monosaccharide composition of EFPA was analyzed by high-performance gel permeation chromatography (HPGPC) and high performance liquid chromatography (HPLC). Lipid accumulation model in HepG2 cells was induced by different concentrations of oleic acid for 24 h,and 10, 30, 100, 300 μg·mL^-1 EFPA and EFPA-R and oleic acid acted on cells for 24 h. CCK-8 was used to evaluate the toxicity of EFPA and EFPA-R to HepG2 cells. After being treated with different concentrations of EFPA and EFPA-R, lipid accumulation in the cells was observed by oil red O staining, and the contents of triglyceride (TG) and cholesterol (TC) in the cells were measured.Results EFPA is a homogeneous polysaccharide with a molecular weight of about 125.8 kDa, containing mannose, glucose, galactose, glucuronic acid and arabinose, with a molar ratio of 1.7∶7.4:1.4∶1.8∶1.0, EFPA-R is containing mannose, glucose, galactose and arabinose, with a molar ratio of 0.8∶10.6∶2.1∶1.0. EFPA and EFPA-R (10, 30, 100, 300 μg·mL^-1) had no significant effect on cell viability after treated cells for 24 h. Compared with control group,the TC and TG content and lipid droplets in oleic acid-induced HepG2 cells in model group was significantly increased (P < 0.01). Compared with the model group, the red lipid droplets in the EFPA intervention groups were significantly reduced, the TC and TG level was significantly decreased (P < 0.05 or P < 0.01). Furthermore, there were no significant changes in the cells after the EFPA-R intervention.Conclusion EFPA could dose-dependently decrease the cholesterol and triglyceride levels in oleic acid-induced HepG2 cells, and significantly inhibit lipid accumulation. The presence of GalA may be a key factor in its inhibition of lipid accumulation in HepG2 cells.

R285.5

吉林省教育厅科学研究项目（JJKH20210988KJ）：基于肠道菌群朝鲜淫羊藿多糖调控NAFLD大鼠肝脏炎症的作用机制，负责人：李波。