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1.佳木斯大学 药学院,黑龙江 佳木斯 154007
2.黑龙江中医药大学 药学院,哈尔滨 150040
[第一作者] 张凯,在读硕士,从事白鲜皮活性成分及药理活性研究,Tel:0454-8616665,E-mail:zhangkai930222@163.com
*赵宏,博士,副教授,从事中药活性成分的结构和活性研究,Tel:0454-8610838,E-mail:0316zh@163.com
纸质出版日期:2020-05-05,
网络出版日期:2020-01-16,
收稿日期:2019-10-08,
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张凯, 赵宏, 张宇, 等. 白鲜皮多糖的分离纯化及抗银屑病作用[J]. 中国实验方剂学杂志, 2020,26(9):157-166.
Kai ZHANG, Hong ZHAO, Yu ZHANG, et al. Purification, Characterization and Anti-psoriasis Effects of Polysaccharides from
张凯, 赵宏, 张宇, 等. 白鲜皮多糖的分离纯化及抗银屑病作用[J]. 中国实验方剂学杂志, 2020,26(9):157-166. DOI: 10.13422/j.cnki.syfjx.20200914.
Kai ZHANG, Hong ZHAO, Yu ZHANG, et al. Purification, Characterization and Anti-psoriasis Effects of Polysaccharides from
目的:
2
对白鲜皮粗多糖(DDP)进行分离纯化,并研究其抗银屑病作用。
方法:
2
DDP经膜分离技术截留相对分子质量小于10 kDa的组分(DDP-UF),采用DEAE-52纤维素柱分离纯化得到4种组分(DDP-UF-1,DDP-UF-2,DDP-UF-3,DDP-UF-4);并通过红外光谱法,高效凝胶渗透色谱法(HPGPC),高效液相色谱法(HPLC),扫描电镜(SEM)测定其理化性质及结构特征。选用咪喹莫特乳膏诱导银屑病小鼠模型,己烯雌酚诱导雌鼠阴道上皮细胞增殖,酶联免疫吸附测定法(ELISA)检测各组小鼠血清白细胞介素(IL)-17与IL-23含量,苏木精-伊红(HE)染色法观察小鼠背部皮肤组织病理变化、阴道上皮细胞有丝分裂指数变化。
结果:
2
DDP-UF-1~4均具有多糖特征吸收峰,DDP-UF-1~4相对分子质量分别为10 948,40 148,32 222,19 943 Da;单糖组成及摩尔比分别为甘露糖-葡萄糖-半乳糖(32.45∶11.35∶8.69),甘露糖-鼠李糖-葡萄糖醛酸-葡萄糖-木糖(25.68∶23.44∶21.62∶18.86∶3.68),甘露糖-鼠李糖-葡萄糖醛酸-半乳糖醛酸-木糖-半乳糖(18.68∶4.61∶3.89∶1.65∶5.36∶6.21),葡萄糖醛酸-半乳糖醛酸-葡萄糖-木糖-半乳糖(11.63∶15.26∶5.32∶2.08∶3.46);扫描电镜(SEM)显示DDP-UF-1~4形态结构为片状或海绵状结构。DDP-UF-1与DDP-UF-3可改善银屑病小鼠背部皮损状态、抑制雌鼠阴道上皮细胞有丝分裂、明显降低血清IL-17,IL-23含量(
P
<
0.05,
P
<
0.01)。
结论:
2
DDP-UF-1与DDP-UF-3均具有良好的抗银屑病作用,其作用可能与抑制IL-23/IL-17信号通路有关。
Objective:
2
To extract crude polysaccharides from
Dictamnus dasycarpus
(DDP) for separation and purification
and study its anti-psoriasis effect.
Method:
2
After interception of DDP with a molecular weight of less than 10 kDa (DDP-UF) using membrane separation technology
four components (DDP-UF-1
DDP-UF-2
DDP-UF-3
DDP-UF-4) were isolated and purified by DEAE-52 cellulose column. Then
the physical and chemical properties and structural characteristics of DDP-UF-1-4 samples were determined by infrared spectroscopy
high performance gel permeability chromatography (HPGPC)
high performance liquid chromatography (HPLC) and scanning electron microscope (SEM). Imiquimod cream was selected to induce mouse models of psoriasis
diethylstilbestrol was used to induce vaginal epithelial cell proliferation in female mice
interleulein-17(IL-17) and IL-23 contents of serum in each mouse group were detected by enzyme-linked immunosorbent assay (ELISA)
and skin tissues of the mouse back and vaginal epithelial cells had mitotic index changes by hematoxylin-eosin staining (HE).
Result:
2
DDP-UF-1-4 all exhibited the characteristic absorption peaks of polysaccharide
and the molecular weights of DDP-UF-1-4 were 10 948
40 148
32 222 and 19 943 Da
respectively. The monosaccharide compositions and mole ratios of DDP-UF-1-4 were mannose-glucose-galactose(32.45∶11.35∶8.69)
mannose-rhamnose-glucuronic acid-glucose-xylose(25.68∶23.44∶21.62∶18.86∶3.68)
mannose-rhamnose-glucuronic acid-galacturonic acid-xylose-galactose(18.68∶4.61∶3.89∶1.65∶5.36∶6.21)
glucuronic acid-galacturonic acid-glucose-xylose-galactose(11.63∶15.26∶5.32∶2.08∶3.46)
respectively. SEM showed that the morphological structures of DDP-UF-1-4 were flaky or spongy. The drug groups of DDP-UF-1 and DDP-UF-3 improved the skin condition of the psoriasis mice back
inhibited mitosis of female vaginal epithelial cells and significantly reduced the contents of IL-17 and IL-23 in serum (
P
<
0.05
P
<
0.01).
Conclusion:
2
Both DDP-UF-1 and DDP-UF-3 have good anti-psoriasis effects
which may be related to the inhibition of IL-23/IL-17 signaling pathway.
白鲜皮多糖银屑病炎症因子小鼠阴道上皮细胞
Dictamnus dasycarpuspolysaccharidespsoriasisinflammatory factormouse vaginal epithelium
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