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目的:从罗氏海盘车体壁、内脏中分别提取粘多糖MP1和MP2,测定其对正常小鼠脾细胞及皂苷作用后的脾细胞免疫活性的影响。方法:罗氏海盘车内脏先经丙酮脱脂,根据正交实验分析确定最佳热水提取条件,然后经热水提取、去蛋白、醇沉得内脏粘多糖MP2。小鼠脾细胞体外培养并分为5组:MP1组、MP2组、皂苷组、MP1+皂苷组及MP2+皂苷组,分别加入相应药物,MTT法分别测定对脾细胞的体外增殖影响。结果:不同浓度MP1和MP2对脾细胞均有一定的促进增殖作用,该作用随多糖剂量的升高先增高后降低;并不同程度地抑制皂苷的细胞毒作用,终浓度为25~40 mg/mL的MP2能逆转皂苷的细胞毒作用,其量效关系是以40 mg/mL(P<0.01)为顶点的抛物线。结论:MP1和MP2均对正常小鼠的免疫功能有促进作用,且以后者效果明显;MP2具有显著的逆转皂苷脾细胞毒作用。
OBJECTIVE: To study the effect of mucopolysaccharide (MP1) and MP2 (MP2) on spleen cells and spleen cells of spleen cells and saponin of normal mice. Methods: The internal organs of Roche seaweed were degreased with acetone firstly, then the optimal hot water extraction conditions were determined by orthogonal experimental analysis, and then extracted by hot water to deproteinize the alcohol to visceral mucopolysaccharide MP2. The splenocytes of mice were cultured in vitro and divided into five groups: MP1 group, MP2 group, saponin group, MP1 + saponin group and MP2 + saponin group. The corresponding drugs were respectively added and the proliferation of spleen cells was determined by MTT assay. Results: Different concentrations of MP1 and MP2 could promote the proliferation of splenocytes, which increased first and then decreased with the increase of polysaccharide dosage. The cytotoxic effects of saponin were inhibited to some extent. The final concentrations of MP1 and MP2 were 25-40 mg / mL of MP2 reversed the cytotoxic effect of saponin. The dose-response relationship was a parabola with apex at 40 mg / mL (P <0.01). CONCLUSION: Both MP1 and MP2 promote the immune function of normal mice, and the latter effect is obvious. MP2 has a significant reversal of splenocyte cytotoxicity.