荔枝核总黄酮对TGF-β_1诱导人肝星状细胞凋亡的影响及机制

目的探讨荔枝核总黄酮（TFL）对转化生长因子β_1（TGF-β_1）诱导的人肝星状细胞（HSC-LX2）凋亡的影响,探讨其相关的分子机制。方法将培养好的HSC-LX2随机分为正常组、TGF-β_1组及150、300、600 mg/L TFL组。正常组常规培养;其他四组接种于含10%FBS的DMEM＋5μg/L TGF-β_1、培养皿中培养24 h后,150、300、600 mg/L TFL组分别加入150、300、600 mg/L TFL继续培养48 h。用Hoechst33258染色法观察各组细胞形态学变化;用流式细胞术检测细胞周期;用FITC Annexin V/PI双染色法检测细胞凋亡情...

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Bibliographic Details
Published in山东医药 Vol. 58; no. 5; pp. 13 - 16
Main Author 曹杰;林丽馨;覃桂金;肖绪华;霍群;赵永忠
Format Magazine Article
LanguageChinese
Published 2018
Subjects
肝纤维化;荔枝核总黄酮;人肝星状细胞;细胞凋亡;转化生长因子β1
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ISSN1002-266X

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Summary:目的探讨荔枝核总黄酮（TFL）对转化生长因子β_1（TGF-β_1）诱导的人肝星状细胞（HSC-LX2）凋亡的影响,探讨其相关的分子机制。方法将培养好的HSC-LX2随机分为正常组、TGF-β_1组及150、300、600 mg/L TFL组。正常组常规培养;其他四组接种于含10%FBS的DMEM＋5μg/L TGF-β_1、培养皿中培养24 h后,150、300、600 mg/L TFL组分别加入150、300、600 mg/L TFL继续培养48 h。用Hoechst33258染色法观察各组细胞形态学变化;用流式细胞术检测细胞周期;用FITC Annexin V/PI双染色法检测细胞凋亡情况;用Western blotting法检测细胞中Toll样受体4（TLR4）、核因子κB（NF-κB）、白细胞介素1受体Ⅰ（IL-1RⅠ）蛋白表达。结果正常组、TGF-β_1组细胞核形态完整,TGF-β_1组细胞增殖活跃;150、300、600 mg/L TFL组可见细胞核固缩、裂解、凋亡小体形成等细胞凋亡形态学变化。随TFL浓度升高,TFL组G0/G1期细胞增多（P〈0.05）,S期细胞减少（P〈0.05）。随TFL浓度升高,TFL对HSC-LX2凋亡具有促进作用,更倾向于促进细胞晚期凋亡。150、300、600 mg/L TFL组TLR4、NF-κB、IL-1RⅠ蛋白表达量低于TGF-β_1组（P均〈0.05）;且随TFL浓度升高,HSC-LX2内TLR4、NF-κB、IL-1RⅠ蛋白表达量逐渐降低（P均〈0.05）。结论 TFL可促进TGF-β_1诱导的HSC-LX2细胞凋亡,尤其促进细胞晚期凋亡;其分子机制可能与降低细胞内TLR4、NF-κB、IL-1RⅠ的表达有关。
Bibliography:Objective To investigate the effects of total flavonoids of semen litchi（ TFL） on the apoptosis of human hepatic stellate cells LX2（ HSC-LX2） induced by transforming growth factor β1（ TGF-β1）,and to explore the related molecular mechanisms. Methods The cultured HSC-LX2 cells were randomly divided into the normal group,TGF-β1 group,and the different concentrations of TFL groups（ 150,300,and 600 mg/L TFL）. Cells in the TGF-β1 group and different concentrations of TFL groups（ 150,300,and 600 mg/L TFL） were cultured in the culture dish with DMEM（ 10% FBS） and TGF-β1（ 5 μg/L）; after 24-hour culture,different concentrations of TFL groups（ 150,300,and 600 mg/L TFL） were added with different concentrations of TFL（ 150,300,and 600 mg/L TFL）; after 48 h,the related indicators were measured. The morphological changes of cells were observed by Hoechst33258 staining; the cell cycle was detected by flow cytometry; FITC Annexin V/PI double staining was used to detect the apoptosis; the expression levels of TLR4,NF-κB,and IL
ISSN:1002-266X