BAPTA-AM抗D-GalN致L-02细胞损伤作用

Protective effects and mechanism of BAPTA-AM on D-galactosmine-induced injury of L-02 cells

探讨了BAPTA-AM对D-氨基半乳糖(D-GalN)诱导的人胚肝细胞(L-02细胞)损伤的影响及其作用机制.加入不同浓度的BAPTA-AM1h后,用D-GalN诱导L-02损伤,通过倒置光学显微镜观察,检测培养液中乳酸脱氢酶(LDH)、谷丙转氨酶(ALT)及谷草转氨酶(AST)的活性和丙二醛(MDA)的含量,MTT法检测细胞活力,Annexin V-EGFP荧光染色统计细胞凋亡率,Ca2+荧光指示剂Fura-2/AM测定细胞内游离钙浓度等方法,评价BAPTA-AM对D-GalN诱导的L-02细胞损伤的影响.实验表明BAPTA-AM能明显抑制D-Gal N引起的L-02细胞培养液中的LDH、ALT、AST和MDA水平的升高,能抑制L-02细胞的凋亡和细胞内游离钙浓度的升高,能明显提高L-02细胞活力.而且BAPTA-AM的效价强度是甘利欣的数百倍.

To study the protective effects and mechanism of BAPTA-AM on D-galactosmineinduced injury of L-02 cells, D-galactosmine was used to induce L-02 cells injure after BAPTA- AM were added to the L-02 cells with various concentrations. The viabilities of L-02 cells （by MTT assay）, the levels of LDH, ALT, AST and MDA in supematant were measured. Cell apoptosis was evaluated by Annexin V-EGFP labeling method. Cytoplasmic free Ca^2＋ concentrations were tested by a Ca^2＋ labeling indicator, Fura-2/AM. The results showed that BAPTA-AM could significantly inhibit the D-galactosmine-induced hepatocyte injury, prevent L-02 cells from increase in the cytoplasm Ca^2＋ concentration and suppress apoptosis. And the potency of BAPTA-AM is hundreds times larger than GAN-LIXIN.