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  

Effects of total flavonoids of semen litchi on TGF-β_1-induced apoptosis of human hepatic stellate cells

作　　者：曹杰 林丽馨 覃桂金 肖绪华[1] 霍群[3] 赵永忠[1] 

机构地区：[1]桂林医学院附属医院,广西桂林541000 [2]广西壮族自治区南溪山医院 [3]桂林医学院

出　　处：《山东医药》2018年第5期13-16,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81360659);广西壮族自治区重点实验室开放课题(16-140-46-17);桂林医学院附属医院自筹经费科研项目(GZZC16-51)

摘　　要：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。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-1 RⅠproteins were detected by Western blotting. Results The nuclear morphology of cells in the normal group and the TGF-β1 group was complete,the proliferation was active in the TGF-β1 group,and the morphological changes of apoptosis including pyknosis,lysis and apoptotic bodies of nucleus were observed in the different concentrations of TFL groups（ 150,300,and 600 mg/L TFL）. Flow cytometry showed that with the increasing TFL concentrations,the cells in the G0/G1 phase increased in the TFL groups（ P 0. 05）,decreased in the S phase（ P 0. 05）; with the increasing TFL concentrations,TFL promoted the apoptosis of HSC-LX2,and tended to promote late apoptotic. In the different concentrations of TFL groups（ 150,300,and 600 mg/L TFL）,TLR4,NF-κB,and IL-1 RⅠ protein expression was lower than that of the TGF-β1 group（ P 0. 05）; with the increase of TFL concentrations,the expression of TLR4,NF-κB,and IL-1 R I protein in HSC-LX2 decreased gradually�

关 键 词：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 

分 类 号：R575[医药卫生—消化系统]