星点设计-效应面法优化枸杞多糖含片的处方并考察其免疫调节作用

Optimization of Prescription of Lycii Fructus Polysaccharide Buccal Tablets by Central Composite Design-response Surface Methodology and Investigation of Its Immunomodulatory Effect

目的:制备枸杞多糖(LBP)含片,并考察其免疫调节作用。方法:以含片的外观、口感、硬度、崩解时限为综合评价指标,采用单因素试验考察LBP提取物用量、填充剂、润湿剂、矫味剂、香精、崩解剂、润滑剂对LBP含片质量的影响,结合星点设计-效应面法优化LBP含片的处方。昆明种小鼠随机分为5组,即LBP含片低(100 mg·kg^-1·d^-1),中(200 mg·kg^-1·d^-1),高(300 mg·kg^-1·d^-1)剂量组,正常组(0.9%生理盐水,300 mg·kg^-1·d^-1)和阳性药组(赐能素组,300 mg·kg^-1·d^-1),计算小鼠免疫器官指数、单核巨噬细胞吞噬指数、血清溶血素抗体水平及迟发型变态反应(DTH)的足趾厚度差,探讨LBP含片的免疫调节作用。结果:LBP含片最优处方为LBP提取物用量80%,糊精-甘露醇(1.2∶1)用量11.5%,甜蜜素-苹果酸(1∶1)用量1%,奶油香精用量0.5%,羧甲基淀粉钠(CMS-Na)用量6.5%,硬质酸镁用量0.5%,80%乙醇适量;在该最优条件下,LBP含片硬度11.83 kg,崩解时限13.21 min,均符合2015年版《中国药典》的相关规定,且制得的含片外观、口感均较好。与正常组比较,LBP含片中、高剂量组可显著增加小鼠胸腺指数、脾脏指数和吞噬指数(P<0.05,P<0.01),LBP各剂量组均可显著增加小鼠血清溶血素抗体水平(P<0.05,P<0.01),LBP含片高剂量组可显著提高小鼠DTH程度(P<0.05)。结论:采用星点设计-效应面法优选所得的LBP含片处方工艺稳定可行,LBP含片可改善正常小鼠的免疫调节功能,为枸杞及LBP的开发利用、临床应用提供了实验依据。

Objective:To prepare Lycii Fructus polysaccharide buccal tablets and investigate its immunomodulatory effect.Method:Taking the appearance,taste,hardness and disintegration time of the tablets as comprehensive evaluation index,based on single factor tests,central composite design-response surface methodology was adopted to optimize the prescription of Lycii Fructus polysaccharide buccal tablets with mass ratio of dextrin to mannitol,mass ratio of cyclamate to malic acid and dosage of sodium carboxymethyl starch(CMS-Na)as factors.Kunming mice were randomly divided into 5 groups,namely the Lycii Fructus polysaccharide buccal tablets low(100 mg·kg^-1·d^-1),medium(200 mg·kg^-1·d^-1)and high(300 mg·kg^-1·d^-1)dose groups,the normal group(0.9%normal saline,300 mg·kg^-1·d^-1)and the positive medicine group(Cinengsu group,300 mg·kg^-1·d^-1).The immunomodulatory effect of the buccal tablets were investigated by calculating immune organ index,monocyte-macrophage phagocytic index,serum hemolysin antibody level,and the voix pedis thickness difference of delayed hypersensitivity(DTH)of mice.Result:Optimal prescription for the buccal tablets was80%of Lycii Fructus extract,11.5%of dextrin-mannitol(1.2∶1),1%of cyclamate-malic acid(1∶1),0.5%of cream essence,6.5%of CMS-Na,0.5%of magnesium stearate,and appropriate amount of 80%ethanol.Under the optimal condition,the hardness of the buccal tablets was 11.83 kg,its disintegration time was 13.21 min,both of which were in line with the relevant provisions of the 2015 edition of Chinese Pharmacopoeia,and the buccal tablets had good appearance and taste.Compared with the normal group,medium and high dose groups of Lycii Fructus polysaccharide buccal tablets significantly increased thymus index,spleen index and phagocytic index of mice(P<0.05,P<0.01),serum hemolysin antibody level of mice was significantly increased in all Lycii Fructus polysaccharide buccal tablets groups(P<0.05,P<0.01),and DTH level of mice was significantly increased in high dose group of Lycii Fructus polysaccharide buccal tablets(P<0.05).Conclusion:The formulation process of the buccal tablets optimized by central composite design-response surface methodology is stable and feasible,and Lycii Fructus polysaccharide buccal tablets can improve the immune regulation function of normal mice,and this study can provide experimental basis for the development,utilization and clinical application of Lycii Fructus and Lycii Fructus polysaccharides.