胆固醇诱导人肝细胞C反应蛋白基因表达

Effects of cholesterol on C-reactive protein gene expression in L-02 cell line and preliminary study on mechanism

目的研究胆固醇对人肝细胞L-02C反应蛋白(C-reactive protein,CRP)基因表达的影响并初步探讨其机制。方法分别以浓度为6.25mg/L,12.5mg/L,25.0mg/L,50.0mg/L的胆固醇作用人肝细胞L-02,采用real-time PCR法检测细胞中CRP mRNA转录情况;Western blot检测细胞内CRP基因表达;免疫比浊法检测细胞培养液中CRP含量,观察胆固醇对L-02细胞CRP基因表达的影响。用荧光分光光度计测定胆固醇刺激细胞后细胞内第二信使钙离子浓度变化情况,初步探讨胆固醇诱导CRP基因表达的分子机制。结果胆固醇能诱导人肝细胞C反应蛋白的表达(P<0.05),其诱导效应随浓度增加而增强;同时细胞内钙离子浓度随胆固醇剂量的增加而增大(P<0.05)。结论胆固醇能诱导人肝细胞L-02CRP基因表达,其机制可能与上调细胞内钙离子浓度有关。

Objective To study the effects of cholesterol on CRP gene expression in L-02 cell line and the relative mechanism. Methods Human hepatocyte cell line L-02 were cultured in vitro. Growth-arrested L-02 cell line were stimulated with different concentration of cholesterol （6.25mg/L, 12.5mg/L, 25.0mg/L, 50.0mg/L）.CRP mRNA were detected by real-time PCR ;intracellular protein by western blot;Secreted CRP by immunoturbidimetry;intracellular Ca^2＋ were detected after L-02 cell line were treated by cholesterol. Results Cholesterol can stimulate CRP gene expression in L-02 （P〈0.05）Compared with the control group, CRP and the concentration of intracellular Ca^2＋ of every group （cholesterel dosage at 6.25mg/L, 12.5mg/L, 25.0mg/L, 50.0mg/L, respectively） significantly increased. Conclusions Cholesterol can induce L- 02 CRP gene expression,which might be related to the concentration of intracellular Ca^2＋.