CAJ | 학술논문

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/L TFL），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。
목적：연구려지핵총황동(TFL)대전화생장인자-β1(TGF-β1)유도적대서간성상세포T6(HSC-T6)세포증식적영향급기상관분자궤제。방법0.25%이단백매소화수집세포，용함10%FBS적DMEM+5μg/L TGF-β1배양액배성단세포현액。（1）MTT법검측세포증식활력。장세포접충우96공배양판중，설TGF-β1조，대조조（함5‰DMSO），TFL80、160、320、640、800조（80、160、320、640、800 mg/L TFL），매조설3개복공。가약24、48、72 h후，채용매표의측정490 nm처각공흡광도（A）치병계산세포억제솔；근거반수억제질량농도（IC50）학정후속실험적약물농도조급약물작용시간。（2）채용PCR화Western blot분별검측HSC-T6세포핵전록인자（NF）-κB、α-평활기기동단백（α-SMA）mRNA화단백적표체。장세포접충우10 cm배양명중，설TGF-β1조，대조조（함5‰DMSO），TFL125、250、500조（125、250、500 mg/L TFL），분조배양48 h후측정。결과동일작용시간점，수착TFL농도적증고，HSC-T6세포적A치기본축점강저，세포억제솔축점상승。TGF-β1조NF-κB、α-SMA mRNA화단백표체량여대조조차이균무통계학의의，TFL125조여TGF-β1조、대조조차이균무통계학의의。수착TFL농도적증고，HSC-T6세포NF-κB、α-SMA mRNA화단백표체량기본축점강저。결론 TFL가억제활화적HSC-T6세포증식，기가능통과억제NF-κB、α-SMA적표체래발휘항간섬유화작용。
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 (5μg/L). (1) MTT method was used to detect the proliferation of HSC-T6 cells. Cells were cultured in 96-well plate and were treated by different concentrations of TFL including TGF-β1 group, the control group (5‰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 time were determined according to half inhibitory concentration (IC50). (2) The expression levels of NF-κB andα-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-κB andα-SMA mRNA between TGF-β1 group and control group. And there were no significant differences in the expressions of NF-κB andα-SMA mRNA between TFL125 group, TGF-β1 group and control group. There was a gradually decrease in the expressions of NF-κB andα-SMA mRNA and protein with the increased concentrations of TFL. Conclusion TFL can inhibit TGF-β1-induced HSC-T6 cell proliferation, which is involved in the inhibited expressions of NF-κB andα-SMA to anti-fibrotic effects in liver fibrosis.