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中文摘要: 通过超声辅助水提法获得刺梨多糖、黑皮鸡枞菌多糖，研究刺梨多糖与黑皮鸡枞菌多糖对转化生长因子‐β1（transforming growth factor‐β1，TGF‐β1）诱导的人肺泡上皮细胞A549（A549 细胞）上皮间质转化（epithelial‐mesenchymal transition，EMT）的影响。采用苯酚‐硫酸法测定多糖含量；将A549 细胞分为正常对照组、模型组、不同剂量多糖干预组。通过MTT 法检测两种多糖对A549 细胞增殖能力的影响，蛋白印迹法检测EMT 标志蛋白表达情况，并结合消化法、流式细胞术分别检测细胞内羟脯氨酸（hydroxyproline，HYP）、活性氧（reactive oxygen species，ROS）含量。结果表明，刺梨多糖、黑皮鸡枞菌多糖含量分别为84.3%、87.6%；与对照组比较，两种多糖对A549 细胞增殖活性呈单向抑制趋势；与模型组相比，两种多糖皆能上调细胞上皮钙黏蛋白（E‐cadherin，E‐cad）表达、下调波形蛋白（vimentin，Vim）及平滑肌肌动蛋白（α‐smooth muscle actin，α‐SMA）表达（P<0.001），降低细胞内HYP 及ROS 含量（P<0.05），且呈现剂量依赖性。综上，刺梨多糖、黑皮鸡枞菌多糖对TGF‐β1 诱导的肺泡上皮细胞间质转化有抑制作用，具有治疗肺纤维化的潜在可能性。
中文关键词: 刺梨多糖 黑皮鸡枞菌多糖 上皮间质转化 特发性肺纤维化 肺泡上皮细胞A549
Abstract:The polysaccharides from Rosa roxburghii Tratt and Oudemansiella raphanipes were extracted via an ultrasonic‐assisted water extraction method.Their effects on transforming growth factor‐β1（TGF‐β1）‐induced epithelial‐mesenchymal transition（EMT）in human alveolar epithelial A549 cells were studied. The polysac‐charide content was quantified by the phenol‐sulfuric acid method. A549 cells were allocated into a normal control group，a model group，and polysaccharide intervention groups with varying doses. The MTT assay was employed to assess the effects of both polysaccharides on A549 cells proliferation. Western blotting was uti‐lized to determine the expression levels of EMT markers，while enzymatic digestion and flow cytometry were ad‐opted to measure intracellular hydroxyproline（HYP）and reactive oxygen species（ROS）levels.Results dem‐onstrated the polysaccharide content was 84.3% and 87.6% in R. roxburghii and O. raphanipes，respectively.Compared with the control group，both polysaccharides exhibited a unidirectional inhibitory trend on A549 cells proliferation. Compared with the model group，both polysaccharides upregulated the expression of E‐cad while downregulating that of vimentin and α‐smooth muscle actin（P<0.001）. Additionally，both polysaccha‐rides lowered intracellular HYP and ROS levels（P<0.05）in a dose‐dependent manner. In conclusion，R. rox?burghii and O.raphanipes polysaccharides inhibited TGF‐β1‐induced EMT in alveolar epithelial cells，demon‐strating potential therapeutic applications for pulmonary fibrosis.
keywords: Rosa roxburghii Tratt polysaccharide Oudemansiella raphanipes polysaccharide epithelial ‐mesenchymal transition idiopathic pulmonary fibrosis human alveolar epithelial A549 cells
文章编号：202512003 中图分类号： 文献标志码：
基金项目:四川省自然科学基金项目（2023NSFSC1251）；四川省农业科学院自主创新科技攻关路径研究项目（2024ZZCX005）；四川省农业科学院原始创新2035 计划项目（YSCX2035‐007）
| 作者 | 单位 |
| 张舒文1，王雯曦1，凌艺菡1，王潇萌1，黄少林1，阴文娅1，黄藩2* | 1.四川大学华西公共卫生学院/华西第四医院,四川成都 610041；2.四川省农业科学院茶叶研究所,四川成都 610066 |
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