摘要
观察BAPTA-AM抗OGD-再灌对HepG-2细胞的损伤作用及其机制.在氧/糖剥夺环境中培养细胞12h后复糖复氧,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,MTT法检测细胞活力,Annexin V-EGFP/PI荧光染色检测细胞死亡率,A23187拮抗实验及Fura-2/AM测定细胞内游离钙浓度等方法,评价BAPTA-AM抗氧/糖剥夺-再灌注致HepG-2细胞损伤的作用.BAPTA-AM能明显抑制OGD-再灌引起的HepG-2细胞培养液中的LDH、ALT、AST水平的升高,抑制HepG-2细胞的坏死和细胞内游离钙浓度的升高,明显提高HepG-2细胞活力,A23187减弱BAPTA-AM保肝作用.BAPTA-AM通过降低细胞内游离钙浓度,明显减轻OGD-再灌对HepG-2细胞的损伤.
To study the protective effects and mechanism of BAPTA-AM on HepG-2 cells against deprivation of 02 and glucose/reperfusion-induced injury, oxygen-glucose deprivation followed by 12 h of reoxygenation-glucose reperfusion was applied to induce HepG-2 cells injure at various concentrations of BAPTA-AM. The viabilities of HepG-2 cells, the levels of ALT, AST, LDH were measured. Cell death was evaluated by Annexin V-EGFP/PI labeling method. Effects of A23187 against BAPTA-AM on viabilities of HepG-2 cells was also measured. Cytoplasmic free Ca2＋ concentrations were tested by a Caz＋ labeling indicator, Fura-2/AM. BAPTA-AM could significantly inhibit deprivation of 02 and glucose/reperfusion -induced hepatocyte injury, prevent HepG-2 cells from cytoplasm Ca2＋ concentrations increase and suppress necrosis. BAPTA-AM could effectively protect HepG-2 cells against deprivation of 02 and glucose/reperfusion induced injury.
出处
《浙江工业大学学报》
CAS
2013年第2期133-137,170,共6页
Journal of Zhejiang University of Technology
