荷花粉多糖显著减轻氟尿嘧啶所致小鼠肠道黏膜屏障损伤

Lotus Bee Pollen Polysaccharides Significantly Relieve Intestinal Mucosal Barrier Damage in Mice Caused by Fluorouracil

目的:探讨不同干燥方法提取的荷花粉多糖(lotus bee pollen polysaccharides,LP)对5-氟尿嘧啶(5-fluorouracil,5-Fu)所致小鼠肠道黏膜损伤及功能失调的改善作用。方法:分别采用热风干燥与真空脉动干燥的方法对荷花粉进行干燥,并提取多糖。建立5-Fu小鼠肠黏膜炎模型,随机分组、给药。炭末推进法测定小鼠的肠推进率,并测量小鼠结肠长度;计算全肠道派氏集合淋巴结(Peyer’s patches,PPs)的面积;取小鼠结肠组织进行病理组织切片观察;生物化学方法检测PPs匀浆上清液中活性氧(reactive oxygen species,ROS)和谷胱甘肽(glutathione,GSH)含量、过氧化氢酶(catalase,CAT)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活力;酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)法检测PPs匀浆上清中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的含量。结果:与正常组相比,5-Fu可显著抑制小鼠肠道的推进功能(P<0.05)。热风干燥与真空脉动干燥样品中提取的LP能减缓5-Fu所致的结肠缩短(P<0.05),显著或极显著增加小鼠肠道PPs的面积(P<0.05或P<0.01);此外,与5-Fu组比较,LP与5-Fu联合应用能明显促进小鼠肠道的推进功能(P<0.05),升高GSH含量、CAT和GSH-Px活力(P<0.01或P<0.05),明显减少TNF-α含量(P<0.05或P<0.01);结肠病理组织切片显示LP能明显减轻5-Fu所致的小鼠结肠黏膜上皮杯状细胞丢失、隐窝深度变浅及炎症细胞浸润。结论:LP能够减轻5-Fu所致小鼠结肠黏膜炎,改善5-Fu所致的肠道传输功能失调,保护5-Fu对肠道的氧化损伤,其中真空脉动干燥样品中提取的花粉多糖效果更好。

Objective： To investigate the protective effects of polysaccharides （LP） extracted from lotus bee pollen driedby different drying methods on intestinal mucosal inflammation and intestinal barrier dysfunctions in mice induced by5-fluorouracil （5-Fu）. Methods： Mouse models of intestinal mucosal inflammation were established by induction with 5-Fuand the model mice were grouped randomly and subjected to drug administration. The carbon-propelling rate was tested,and colon length and Peyer’s patches （PPs） area were measured. The colon tissue was stained with hematoxylin-eosin （HE）for histological section observation. Reactive oxygen species （ROS） and glutathione （GSH） contents, and catalase （CAT） andGSH-Px activities in the supernatant of PPs homogenate were evaluated by biochemical assays. Tumor necrosis factor-α （TNF-α）in the supernatant of colon tissue homogenate was determined through ELISA. Results： Compared with the normal group,intraperitoneal injection of 5-Fu to mice significantly inhibited small intestinal transit （P 〈 0.05）. LP extracted from lotus bee pollendried by hot air or vacuum pulsed drying improved colon shortening caused by 5-Fu （P 〈 0.05） and resulted in a significant or highlysignificant increase of PPs area （P 〈 0.05 or P 〈 0.01）. Moreover, compared with the 5-Fu-induced model group, LP combined with 5-Fu could significantly promote intestinal transit （P 〈 0.05）, increase GSH content, and CAT and GSH-Px activities （P 〈 0.01 orP 〈 0.05）, and decrease obviously TNF-α contents （P 〈 0.05 or P 〈 0.01）. HE staining results showed that LP could significantlyreduce the loss of colonic mucosa epithelium goblet cells, inflammatory cell infiltration and more shallow crypt depth inducedby 5-Fu. Conclusion： LP could relieve intestinal mucosal damage, improve intestinal transit dysfunction and protect againstintestinal oxidative damage in mice induced by 5-Fu. The polysaccharides extracted from lotus bee pollen subjected to vacuumpulsed drying exhibited better activity than the polysaccharides extracted from hot air dried lotus bee pollen.