[1]赵秋芳,汪雪梅,李 强,等.蒺藜皂苷对Graves眼病眼眶组织纤维化的作用机制研究[J].陕西中医,2024,(11):1476-1480,1485.[doi:DOI:10.3969/j.issn.1000-7369.2024.11.007]
　ZHAO Qiufang,WANG Xuemei,LI Qiang,et al.Study on mechanism of terrestroside saponins on fibrosis of eye frame tissue in Graves ophthalmopathy[J].,2024,(11):1476-1480,1485.[doi:DOI:10.3969/j.issn.1000-7369.2024.11.007]
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蒺藜皂苷对Graves眼病眼眶组织纤维化的作用机制研究
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《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2024年11期
页码:
1476-1480,1485
栏目:
基础研究
出版日期:
2024-11-05

文章信息/Info

Title:
Study on mechanism of terrestroside saponins on fibrosis of eye frame tissue in Graves ophthalmopathy
作者:
赵秋芳; 汪雪梅; 李 强; 张 唯
(陕西中医药大学附属医院眼科,陕西 咸阳 712021)
Author(s):
ZHAO Qiufang; WANG Xuemei; LI Qiang; ZHANG Wei
(Department of Ophthalmology,Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712021,China)
关键词:
Graves眼病;  蒺藜皂苷;  磷酸肌醇3激酶;  蛋白激酶B;  组织纤维化
Keywords:
Graves ophthalmopathy;  Terrestroside;  PI3K/AKT;  Tissue fibrosis
分类号:
R 856.77
DOI:
DOI:10.3969/j.issn.1000-7369.2024.11.007
文献标志码:
A
摘要:
目的:研究蒺藜提取物蒺藜皂苷对Graves眼病眼眶组织纤维化的作用及其机制。方法:采用人hTSHR 重组质粒结合电穿孔免疫 BALB/c 雌性小鼠的方法构建Graves 眼病模型,分为对照组、模型组、蒺藜皂苷组。RT-PCR、免疫荧光法观察三组小鼠眼眶组织α-SMA 、Collagen Ⅰ和Collagen Ⅲ水平。采用马松和天狼星红染色观察三组小鼠眼眶组织细胞外基质(ECM)沉积水平; 采用ELISA检测三组小鼠血清PDGF、INF-γ和TGF-β1水平。TGF-β1刺激原代分离小鼠眼球后成纤维细胞,对照组PBS给药处理。蒺藜皂苷分别设置5、10、20、40、80 μg/ml浓度进行给药。CCK-8检测不同组细胞增殖能力变化; RT-PCR、免疫荧光观察不同组细胞α-SMA、Collagen Ⅰ和Collagen Ⅲ水平; ELISA检测不同组细胞上清中PDGF、INF-γ水平; Western blotting检测不同组细胞PI3K/AKT和Smad2通路变化。结果:蒺藜皂苷能抑制小鼠眼眶组织α-SMA 、Collagen Ⅰ和Collagen Ⅲ以及炎症因子PDGF、INF-γ、TGF-β1的表达。蒺藜皂苷显著抑制小鼠眼眶组织ECM的沉积。蒺藜皂苷抑制TGF-β1刺激后小鼠眼球后成纤维细胞增殖,α-SMA、Collagen Ⅰ、Collagen Ⅲ、PDGF、INF-γ的分泌。蒺藜皂苷能抑制纤维化经典通路PI3K/AKT和Smad2磷酸化。结论:蒺藜皂苷能够抑制PI3K/AKT和Smad2通路,阻滞Graves 眼病眼球组织纤维化,可有效治疗Graves 眼病。
Abstract:
Objective:Study on effect and mechanism of terrestroside(TD)in intervening in fibrosis of Graves eye disease eye frame tissue.Methods:Human hTSHR recombinant plasmid combined with electroporation immunize BALB/c female mice to construct Graves ophthalmopathy model.Divided into control group,model group and tribulus saponin treatment group.RT-PCR and immunofluorescence were used to observe levels of α-SMA,Collagen Ⅰ and Collagen Ⅲ in orbital tissues of the three groups.Masson and Sirius red staining observed extracellular matrix(ECM)deposition levels in orbital tissues of three groups.The three groups were detected by ELISA mouse serum PDGF,INF-γ and TGF-β1 levels.TGF-β1 stimulates primary isolated mouse eye fibroblasts,the control group were treated with PBS.The control group were treated with PBS administration.Terrestroside were administered at concentrations of 5,10,20,40,80 μg/ml,respectively.Proliferation ability of cells in different groups were detected by CCK-8.RT-PCR and immunofluorescence were used to observe levels of α-SMA,Collagen Ⅰ and Collagen Ⅲ in the three groups.ELISA were used to detect levels of cells in the groups,and levels of PDGF,and INF-γ in the serum.Western blotting was used to detect changes of PI3K/AKT and Smad2 pathways in the three groups of cells.Results:TD significantly inhibit expression of α-SMA,Collagen Ⅰ and Collagen Ⅲ and inflammatory factors PDGF,INF-γ and TGF-β1 in mouse eye frame tissues.TD significantly inhibit the deposition of ECM in mouse eye frame tissues.Terrestroside significantly inhibit proliferation of posterior fibroblasts in mice after TGF-β1 stimulation,and secretion of α-SMA,Collagen Ⅰ and Collagen Ⅲ,PDGF and INF-γ.TD significantly inhibit fibrosis classics pathway PI3K/AKT and Smad2 phosphorylation.Conclusion:Terrestroside inhibit PI3K/AKT and Smad2 pathways and block fibrosis of eyeball tissue of Graves' ophthalmopathy,effectively treating Graves' ophthalmopathy.

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备注/Memo

备注/Memo:
基金项目:陕西省重点研发计划项目(2017SF-350)
更新日期/Last Update: 2024-11-11