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黄芪多糖对肝窦内皮细胞纳米级力学特性的影响:基于原子力显微镜的研究

Effects of Astragalus Polysaccharide on Mechanical Characterization of Liver Sinusoidal Endothelial Cells by Atomic Force Microscopy at Nanoscale.

作者信息

Lu Wen-Liang, Li Juan-Mei, Yang Jiao, Xu Chen-Guang, Zhang Sha-Sha, Yan Jie, Zhang Ting-Ting, Zhao Hui-Hui

机构信息

Department of Liver Diseases, Guang'anmen Hospital, China Academy of Chinese Medical Sciences, Beijing, 100053, China.

Department of Traditional Chinese Medicine, The First People's Hospital of Huzhou, Huzhou, Zhejiang Province, 313000, China.

出版信息

Chin J Integr Med. 2018 Jun;24(6):455-459. doi: 10.1007/s11655-017-2964-0. Epub 2017 Jul 25.

Abstract

OBJECTIVE

To study the effects of Astragalus polysaccharide (APS), the primary effective component of the Chinese herb medicine Astragalus membranaceus (frequently used for its anti-hepatic fibrosis effects), on nanoscale mechanical properties of liver sinusoidal endothelial cells (SECs).

METHODS

Using endothelial cell medium as the control, 5 experimental groups were established utilizing different concentrations of APS, i.e. 12.5, 25, 50, 100, and 200 μg/mL. By using atomic force microscopy along with a microcantilever modified with a silicon dioxide microsphere as powerful tools, the value of Young's modulus in each group was calculated. SAS 9.1 software was applied to analyze the values of Young's modulus at the pressed depth of 300 nm. Environmental scanning electron microscopy was performed to observe the surface microtopography of the SECs.

RESULTS

The value of Young's modulus in each APS experimental group was significantly greater than that of the control group: as APS concentration increased, the value of Young's modulus presented as an increasing trend. The difference between the low-concentration (12.5 and 25 μg/mL) and high-concentration (200 μg/mL) groups was statistically significant (P<0.05), but no significant differences were observed between moderateconcentration (50 and 100 μg/mL) groups versus low- or high-concentration groups (P>0.05). Surface topography demonstrated that APS was capable of increasing the total area of fenestrae.

CONCLUSIONS

The values of Young's modulus increased along with increasing concentrations of APS, suggesting that the stiffness of SECs increases gradually as a function of APS concentration. The observed changes in SEC mechanical properties may provide a new avenue for mechanistic research of anti-hepatic fibrosis treatments in Chinese medicine.

摘要

目的

研究中药黄芪的主要有效成分黄芪多糖(APS,常用于抗肝纤维化)对肝窦内皮细胞(SECs)纳米级力学性能的影响。

方法

以内皮细胞培养基作为对照,利用不同浓度的APS(即12.5、25、50、100和200μg/mL)建立5个实验组。通过使用原子力显微镜以及用二氧化硅微球修饰的微悬臂梁作为有力工具,计算每组的杨氏模量值。应用SAS 9.1软件分析在300nm压入深度时的杨氏模量值。进行环境扫描电子显微镜观察SECs的表面微观形貌。

结果

每个APS实验组的杨氏模量值均显著高于对照组:随着APS浓度增加,杨氏模量值呈上升趋势。低浓度(12.5和25μg/mL)组与高浓度(200μg/mL)组之间的差异具有统计学意义(P<0.05),但中等浓度(50和100μg/mL)组与低浓度或高浓度组之间未观察到显著差异(P>0.05)。表面形貌表明APS能够增加窗孔的总面积。

结论

杨氏模量值随APS浓度增加而升高,表明SECs的硬度随着APS浓度的增加而逐渐增加。观察到的SECs力学性能变化可能为中药抗肝纤维化治疗的机制研究提供一条新途径。

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