芹菜素抑制人良性胆管瘢痕成纤维细胞的增殖迁移并诱导其凋亡和机制  

Inhibition Effect of Apigenin on Proliferation,Migration,and Induces Apoptosis of Human Benign Bile Duct Scar Fibroblasts

作　　者：胡晟 张小文[1] HU Sheng;ZHANG Xiaowen(Department of Hepatobiliary and Pancreatic Surgery,the Second Affiliated Hospital of Kunming Medical University,Kunming 650101,China)

机构地区：[1]昆明医科大学第二附属医院肝胆胰外科二病区,昆明650101

出　　处：《医药导报》2021年第8期997-1004,共8页Herald of Medicine

基　　金：国家自然科学基金资助项目(81760430)。

摘　　要：目的研究芹菜素对体外原代培养的人良性胆管瘢痕成纤维细胞(HBBDSF)的增殖、迁移、凋亡的影响,以及观察对细胞中的转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白(CollagenⅠ)和Ⅲ型胶原蛋白(CollagenⅢ)的表达水平的影响。方法体外原代培养HBBDSF并鉴定,细胞计数试剂盒(CCK-8)法和BeyoClick TM EdU-488细胞增殖检测试剂盒(EDU-488)法检测芹菜素对细胞增殖的影响;细胞划痕实验检测芹菜素对细胞迁移能力的影响;流式细胞仪检测芹菜素诱导细胞凋亡情况;细胞免疫荧光和Western blotting检测经芹菜素干预后细胞中TGF-β1、α-SMA、Collagen I和CollagenⅢ蛋白的表达情况。结果波形蛋白和角蛋白阳性表达,证明为HBBDSF。CCK-8法测芹菜素对细胞的IC 50值为22.75μmol·L^(-1),R 2=0.797。CCK-8和EDU-488法显示经芹菜素处理后细胞增殖能力明显抑制(P<0.01);细胞划痕实验显示芹菜素对细胞迁移能力明显抑制(P<0.01);芹菜素可诱导细胞凋亡的发生(P<0.01);芹菜素抑制细胞中TGF-β1、α-SMA、Collagen I和CollagenⅢ蛋白的表达(P<0.05)。结论芹菜素可通过抑制TGF-β1表达来抑制HBBDSF的增殖、迁移能力,并诱导凋亡;同时细胞中的TGF-β1、α-SMA、Collagen I和CollagenⅢ蛋白的表达水平明显降低。Objective To study the effect of Apigenin on the proliferation,migration and apoptosis of HBBDSF in vitro,and to observe the effect of Apigenin on TGF-β1,α-SMA,Collagen I and CollagenⅢprotein expression levels.Methods In vitro primary culture of HBBDSF and identified,Cell Counting Kit-8 method and BeyoClick EdU Cell Proliferation Kit with Alexa Fluor 488 method to detect the effect of Apigenin on cell proliferation;The effect of Apigenin on cell migration was detected by cell scratch test.The apoptosis induced by Apigenin was detected by flow cytometry.Detection of TGF-β1,α-SMA,Collagen I and CollagenⅢprotein expression levels in cells after Apigenin incubation was carried out by cell immunofluorescence and Western Boltting.Results Vimentin and Keratin were positively expressed,demonstrating HBBDSF occurred in cells.The IC 50 value measured by CCK-8 method was 22.75μmol·L^(-1),R 2=0.797.CCK-8 and EDU-488 methods showed significant inhibition of cell proliferation after Apigenin treatment(P<0.01).Cell scratch test showed Apigenin significantly inhibited cell migration(P<0.01).Apigenin can induce apoptosis(P<0.01).Celery,inhibits cell of TGF-β1,α-SMA,Collagen I and CollagenⅢprotein expression(P<0.05).Conclusion Apigenin can inhibit proliferation and migration of HBBDSF and induce apoptosis by inhibiting TGF-β1.At the same time in cells TGF-β1,α-SMA,Collagen I and CollagenⅢprotein expression level decreased.

关 键 词：芹菜素 良性胆管狭窄 TGF-Β信号通路 成纤维细胞 

分 类 号：R657.46[医药卫生—外科学]