荔枝核总黄酮对TGF-β1诱导的大鼠肝星状细胞内NF-κB、α-SMA表达的影响  

Effects of the total flavone of litchi chinensis sonn on expressions of NF-κB and α-SMA in TGF-β1 activated rat hepatic stellate cells

作　　者：刘燕秀[1] 赵永忠[1] 李彩[1] 董勇[1] 覃桂金 成秋宸[1] 郑清华[1] 

机构地区：[1]桂林医学院附属医院消化内科,541001

出　　处：《天津医药》2015年第11期1258-1262,共5页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(81360659);广西自然科学基金项目(2012GXNSFAA053105)

摘　　要：目的研究荔枝核总黄酮(TFL)对转化生长因子-β1(TGF-β1)诱导的大鼠肝星状细胞T6(HSC-T6)细胞增殖的影响及其相关分子机制。方法 0.25%胰蛋白酶消化收集细胞,用含10%FBS的DMEM+5μg/L TGF-β1培养液配成单细胞悬液。(1)MTT法检测细胞增殖活力。将细胞接种于96孔培养板中,设TGF-β1组,对照组(含5‰DMSO),TFL80、160、320、640、800组(80、160、320、640、800 mg/L TFL),每组设3个复孔。加药24、48、72 h后,采用酶标仪测定490 nm处各孔吸光度(A)值并计算细胞抑制率;根据半数抑制质量浓度(IC50)确定后续实验的药物浓度组及药物作用时间。(2)采用PCR和Western blot分别检测HSC-T6细胞核转录因子(NF)-κB、α-平滑肌肌动蛋白(α-SMA)mRNA和蛋白的表达。将细胞接种于10 cm培养皿中,设TGF-β1组,对照组(含5‰DMSO),TFL125、250、500组(125、250、500 mg/LTFL),分组培养48 h后测定。结果同一作用时间点,随着TFL浓度的增高,HSC-T6细胞的A值基本逐渐降低,细胞抑制率逐渐上升。TGF-β1组NF-κB、α-SMA mRNA和蛋白表达量与对照组差异均无统计学意义,TFL125组与TGF-β1组、对照组差异均无统计学意义。随着TFL浓度的增高,HSC-T6细胞NF-κB、α-SMA mRNA和蛋白表达量基本逐渐降低。结论 TFL可抑制活化的HSC-T6细胞增殖,其可能通过抑制NF-κB、α-SMA的表达来发挥抗肝纤维化作用。Objective To investigate the effects of total flavonoids of litchi chinensis sonn （TFL） on cell proliferation and the molecular mechanism in rat hepatic stellate cells （HSC-T6） activated by growth factor-β1 （TGF-β1）. Methods HSC-T6 cells were treated by 0.25% Trypsin-EDTA and then were digested into single cell suspension by DMEM （10% FBS included）, which were mixed with TGF-β1 （5 μg/L）. （1） MTr method was used to detect the proliferation of HSC-T6 cells. Ceils were cultured in 96-well plate and were treated by different concentrations of TFL including TGF-131 group, the control group （5%o DMSO included）, and different concentrations of TFL groups （80, 160, 320, 640 and 800 mg/L TFL）. Each group has three wells. The absorbance （A） value was measured by enzyme standard meter at the 490 nm wavelength after 24 h, 48 h and 72 h treatment. The cell inhibitory rate was calculated. The subsequent experimental drug concentration and drug treatment timewere determined according to half inhibitory concentration （IC50）. （2） The expression levels of NF-κB and α-SMA mRNA were detected by PCR （for mRNA） and Western blot assay （for protein）. Cells were cultured in the 10 cm culture dish and were divided into different TGF-β1 groups, including TGF-β1 group, the control group （5‰DMSO included）, and different concentrations of TFL groups （125, 250 and 500 mg/L TFL）. After 48 h, related indicators were measured. Results At the same treatment time point, with the increased concentrations of TFL, A values were gradually decreased, and the cell inhibitory rates were gradually increased. There were no significant differences in the expressions of NF-KB and α-SMA mRNA between TGF-β1 group and control group. And there were no significant differences in the expressions of NF-KB and α-SMA mRNA between TFL125 group, TGF-β1 group and control group. There was a gradually decrease in the expressions of NF-κB and α-SMA mRNA and protein with the increased concentrations

关 键 词：黄酮类 肝硬化 大鼠 NF—κB 荔枝核总黄酮 肝星状细胞 Α-平滑肌肌动蛋白 

分 类 号：R575[医药卫生—消化系统]