荔枝核总黄酮对体外活化肝星状细胞-T6增殖、凋亡及TLR4/NF-κB途径的影响  

Effects of total flavonoids of semen litchi on proliferation, apoptosis and TLR4/NF-κB pathway of activated HSC-T6 in vitro

作　　者：韦铮武[1] 李彩[1] 林丽馨 肖绪华[1] 刘燕秀[1] 董勇[1] 覃桂金 赵永忠[1] 

机构地区：[1]桂林医学院附属医院,桂林541001 [2]广西壮族自治区南溪山医院,桂林541004

出　　处：《中华中医药杂志》2016年第12期5214-5218,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81360659);广西自然科学基金项目(No.2012GXNSFAA053105);广西重点实验室开放课题项目(No.16-140-46-17)~~

摘　　要：目的:研究荔枝核总黄酮(TFL)在体外对转化生长因子-β1(TGF-β1)活化的大鼠肝星状细胞(HSC-T6)增殖、凋亡及TLR4/NF-κB途径的影响,探究该药抗肝纤维化的作用机制。方法:体外培养HSC-T6,用TGF-β1刺激24h后以不同浓度的TFL干预24、48、72h,用MTT法检测TFL对HSC-T6增殖的影响,Hoechst 33258荧光染色观察凋亡细胞形态学变化,采用流式细胞术、荧光定量PCR检测TFL对细胞凋亡及α-SMA、TLR4、NF-κB、Survivin m RNA表达水平的影响。结果:TFL以时间和浓度依赖性抑制活化的HSC-T6增殖,并以作用48h最为显著。TFL可促进细胞凋亡,下调HSC-T6中α-SMA、TLR4、NF-кB和Survivin m RNA表达水平,并具有浓度依赖性。同时Hoechst 33258染色显示TFL各组细胞可见细胞核固缩、边聚和裂解、凋亡小体形成等凋亡形态学变化。结论:TFL在体外可以抑制活化的HSC-T6增殖,促进细胞凋亡,其机制可能与降低活化的HSC-T6内TLR4、NF-кB和Survivin m RNA的表达有关。Objective： To study the effects of total flavonoids of Semen Litchi（TFL） on the proliferation, apoptosis and TLR4/NF-κB pathway of activated rat hepatic stellate cells（HSC-T6） in vitro, and to explore its mechanism of anti-hepatic fibrosis. Methods： HSC-T6 cells were cultured and stimulated by TGF-β1 for 24 h in vitro, then, different concentrations of TFL were given for 24, 48, 72 h. MTT method was used to detect the effect of TFL on proliferation of HSC-T6. Fluorescence staining was used for observing the morphological changes of apoptotic cells. The cell apoptosis was detected by flow cytometry（FCM）. The expression levels of α-SMA, TLR4, NF-κB and Survivin m RNA were measured by Real-Time PCR. Results： TFL could inhibit the proliferation of activated HSC-T6 in a time-dependent and concentration-dependent manner and the effect of 48 h was the most obvious. TFL could promote cell apoptosis and decrease the expressions of α-SMA, TLR4, NF-κB and Survivin m RNA in a concentration-dependent manner. Hoechst 33258 staining showed that TFL cells were found morphological changes of apoptosis, such as nuclear fragmentation, karyopyknosis, chromosomal condensation, cell detachment and so on. Conclusion： In vitro, TFL could inhibit the proliferation of activated HSC-T6 and promoting apoptosis. Its mechanism may be related to the decreasing of TLR4, NF-κB and Survivin m RNA in activated HSC-T6.

关 键 词：荔枝核总黄酮 肝星状细胞 增殖 凋亡 TLR4/NF-κB通路 

分 类 号：R285[医药卫生—中药学]