枸杞多糖及其硫酸酯体外免疫及抗肿瘤活性

Immunomodulatory and antitumor activities of Lycium barbarum polysaccharides in vitro and their sulfates

目的:研究枸杞多糖及其硫酸酯体外免疫和抗肿瘤活性。方法:分别采用水提一步醇沉法和分步醇沉法提取不同的多糖LBPS30、LBPS70和LBPSt,纯化后用氯磺酸-吡啶法修饰得到3种硫酸化枸杞多糖s LBPS30、s LBPS70和s LBPSt。采用MTT法检测多糖对小鼠脾淋巴细胞增殖和食管癌EC109细胞抑制的影响,ELISA法检测细胞上清液中TNF-α、IL-2、IFN-γ和IL-24的含量,划痕法检测细胞迁移情况。结果:s LBPSt和s LBPS30的淋巴细胞增殖率和不同时间点对EC109细胞的抑制率都较高;s LBPSt组细胞上清液中4种淋巴细胞因子含量均显著高于细胞对照组(P<0.05);s LBPSt组细胞迁移数显著低于其余各组(P<0.05)。结论:硫酸化修饰能显著增强体外脾淋巴细胞增殖,促进淋巴细胞四种细胞因子的分泌,并能抑制EC109细胞生长,降低EC109细胞迁移速率,且与硫酸基含量有一定的关系。s LBPSt和s LBPS30总体效果较好。

Aim： To screen the effective Lycium barbarum polysaccharides （ LBPSs ） according to researching the effects of immunity and antitumor activity on LBPSs and their sulfates in vitro .Methods：Three types of LBPSs , LBPS30 ,LBPS70 and LBPSt were extracted by water decoction and stepwise or one-step ethanol precipitation methods and further modified using the chlorosulfonic acid-pyridine method based on previous experiments , yielding three sulfated LBPSs （ sLB-PSs）, sLBPSt, sLBPS30, sLBPS70, respectively.The effects of six polysaccharides spleen lymphocyte proliferation and their antitumor activities against EC109 cells in vitro were evaluated by MTT method .The contents of TNF-α, IL-2, IFN-γand IL-24 in cell supernatant were determined by ELISA method .Cell migration model was induced by scratch method and the migrating cells were counted to observe the effects of polysaccharides on EC 109 cell migration inhibition .Results：The lymphocyte proliferation ratios and inhibition ratio against EC 109 of sLBPSt and sLBPS30 group at different time points were higher than those of other groups , the contents of four cytokines of sLBPSt group were significantly higher than those of the control group（P〈0.05）, the migrating cells of sLBPSt group were also fewer than those of other group （P〈0.05）. Conclusion：sLBPSs could increase conA-activated lymphopoiesis , promote the secretion of TNF-α, IL-2, IFN-γand IL-24.Moreover, they could inhibit the EC109 cells′viability and reduce the rates of tumor cell migration in vitro , indica-ting that sulfated modification enhanced the immunoregulatory and antitumor activities of LBPSs .sLBPS30 and sLBPSt showed the best performance , could be used as materials for further research .