三叶苷对人正常肝细胞L-02增殖作用的研究

Effect of trilobatin on the proliferation in human normal hepatocyte L-02

目的探究三叶苷(TLB)促进人正常肝细胞L-02的增殖作用。方法人正常肝细胞L-02经不同浓度的TLB处理后,通过MTT法测定TLB对L-02细胞活力的影响,采用BrdU ELISA法和EDU荧光染色法分别观察L-02细胞DNA的含量和细胞增殖的改变,通过免疫印迹法分析细胞周期相关调节蛋白的表达。结果TLB(25、50、100μM)作用L-02细胞72 h能够明显增加L-02的细胞活力,并明显增加BrdU含量及EDU的荧光强度,同时TLB能够明显下调L-02细胞中HNF4α的蛋白表达,上调CDK4、cyclin D1、HGF的蛋白表达,并增加c-Met的磷酸化水平和其下游Akt、Erk1/2的磷酸化水平。结论TLB对人正常肝细胞L-02具有促增殖作用,其机制可能与调控HGF/c-Met信号途径有关。

Objective To explore the proliferation effect of trilobatin(TLB)on human normal hepatocyte L-02.Methods L-02 cells were treated with various concentrations of TLB,and the cell viability was detected using MTT assay.Then,DNA content and cell proliferation were observed by BrdU ELISA and EDU staining,respectively.Furthermore,the expressions of cell cycle related proteins were determined using Western blot analysis.Results The results showed that TLB(25,50,100μM)significantly increased the L-02 cell viability within 72 h.Moreover,TLB markedly increased the BrdU level and EDU fluorescence intensity.Furthermore,TLB down-regulated the protein expression of HNF4α,up-regulated the protein expressions of CDK4,cyclin D1 and HGF,as well as augmented the phosphorylation levels of c-Met,Akt and Erk1/2.Conclusion TLB promoted the proliferation of L-02 cells in a dosedependent manner,and its underlying mechanism might be related to regulation of the HGF/c-Met signaling pathway.