CAJ | 학술논문

目的：研究荔枝核总黄酮( TFL)在体外对活化的大鼠肝星状细胞( HSC-T6)的增殖抑制作用及 Toll 样受体4(TLR4)表达的影响。方法使用转化生长因子β1(TGF-β1)(5 ng/ml)激活HSC-T6,显微镜下观察HSC-T6和激活的HSC-T6形态变化；CCK8试剂盒( CCK8)法观察 TFL 作用24、48、72 h后活化的HSC-T6的增殖情况；逆转录聚合酶链反应( RT-PCR)检测HSC-T6中TLR4的表达；流式细胞术检测细胞周期和细胞凋亡。结果显微镜下观察HSC-T6和激活的HSC-T6形态有明显变化；CCK8结果显示TFL可以抑制HSC-T6的增殖( P<0．05),并且随着TFL浓度和作用时间增加,HSC-T6增殖明显下降,呈浓度依赖性；RT-PCR结果显示随着TFL浓度增加,HSC-T6中TLR4表达下降,与实验对照组比较差异有统计学意义( P<0．05)；流式细胞仪检测结果表明TFL阻滞细胞于S期,G0/G1期细胞减少( P<0．05),S期细胞增加(P <0．05),促进细胞的凋亡(P <0．05),呈浓度依赖性。结论 TFL可以抑制 HSC-T6的增殖,其作用机制可能是通过降低TLR4在HSC-T6中的表达,阻滞其细胞增殖和诱导细胞凋亡,达到抗肝纤维化效果。
목적：연구려지핵총황동( TFL)재체외대활화적대서간성상세포( HSC-T6)적증식억제작용급 Toll 양수체4(TLR4)표체적영향。방법사용전화생장인자β1(TGF-β1)(5 ng/ml)격활HSC-T6,현미경하관찰HSC-T6화격활적HSC-T6형태변화；CCK8시제합( CCK8)법관찰 TFL 작용24、48、72 h후활화적HSC-T6적증식정황；역전록취합매련반응( RT-PCR)검측HSC-T6중TLR4적표체；류식세포술검측세포주기화세포조망。결과현미경하관찰HSC-T6화격활적HSC-T6형태유명현변화；CCK8결과현시TFL가이억제HSC-T6적증식( P<0．05),병차수착TFL농도화작용시간증가,HSC-T6증식명현하강,정농도의뢰성；RT-PCR결과현시수착TFL농도증가,HSC-T6중TLR4표체하강,여실험대조조비교차이유통계학의의( P<0．05)；류식세포의검측결과표명TFL조체세포우S기,G0/G1기세포감소( P<0．05),S기세포증가(P <0．05),촉진세포적조망(P <0．05),정농도의뢰성。결론 TFL가이억제 HSC-T6적증식,기작용궤제가능시통과강저TLR4재HSC-T6중적표체,조체기세포증식화유도세포조망,체도항간섬유화효과。
Objective To investigate the anti-proliferative effect of total flavone of litchi chinensis sonn ( TFL) on the activated hepatic stellate cells ( HSC-T6 ) in vitro and the influence on Toll-like receptors 4 ( TLR4 ) . Methods HSC-T6 were activated with TGF-β(5 ng/ml) and then divided into the following groups: blank control, experi-mental control, five experimental groups. Different morphological changes of HSC-T6 and HSC-T6 were observed under the microscope. CCK8 was used to observe the proliferation of three kinds of the activated HSC-T6 which had been respectively affected by TFL for 24, 48 and 72 h. The expressions of TRL4 were detected by RT-PCR ( for mRNA) . The cell cycle and cell apoptosis were detected by flow cytometry ( FCM) . Results There were obvious morphological changes of HSC-T6 and HSC-T6 under the microscope. The results of CCK8 showed that TFL could inhibit the proliferation of HSC-T6 ( P<0. 01 ) , and HSC-T6 decreased significantly with the increase of the con-centration and action time of TFL. It could be found in the results of RT-PCR that expressions of TLR4 of HSC-T6 declined with the increase of TFL concentration. There were visible differences compared with the experimental control (P<0. 01). The test of FCM showed that TFL retarded the cells during PeriA S, G0/G1cells reduce (P<0. 01) and then promoted the apoptosis of cells (P<0. 01). So it depended on the concentration of TFL. Conclu-sion TFL can inhibit the proliferation of HSC-T6 , whose mechanism is to inhibit the proliferation of cells and in-duce the cell apoptosis by reducing the expressions of TLR4 in HSC-T6 , so as to have the effect of anti-liver fibro-sis.