核因子-κB在左卡尼汀抑制肝细胞氧化损伤中的作用

发布时间：2018-09-11 07:44

【摘要】：目的研究左卡尼汀(L-carnitine)抑制过氧化氢(hydrogen peroxide,H_2O_2)诱导的肝细胞损伤作用是否与核因子-κB(nuclear factor-kappa B,NF-κB)有关。方法 CCK-8及乳酸脱氢酶(lactate dehydrogenase,LDH)释放法测定核因子-κB抑制剂对过氧化氢诱导HL7702肝细胞损伤的影响;蛋白质印迹法检测左卡尼汀对过氧化氢损伤HL7702肝细胞核因子-κB表达的影响;细胞免疫荧光染色法测定核因子-κB的核易位情况;凝胶迁移实验(electrophoretic mobility shift assay,EMSA)分析核因子-κB的DNA结合活性。结果与过氧化氢损伤组相比,核因子-κB抑制剂组细胞活性明显升高,乳酸脱氢酶释放率降低(P0.05,P0.01);过氧化氢损伤后细胞核核因子-κB表达增加,左卡尼汀预孵育对过氧化氢诱导的核因子-κB核转位具有明显抑制作用(P0.01);左卡尼汀对过氧化氢诱导的核因子-κB DNA结合活性增加也有抑制作用。结论左卡尼汀通过抑制核因子-κB活性而抑制过氧化氢诱导的肝细胞损伤。
[Abstract]:Aim to investigate whether the inhibitory effect of levocarnitine (L-carnitine) on hepatocyte damage induced by hydrogen peroxide (hydrogen peroxide,H_2O_2) is related to nuclear factor- 魏 B (nuclear factor-kappa Bmf NF- 魏 B. Methods CCK-8 and lactate dehydrogenase (lactate dehydrogenase,LDH) release assay were used to determine the effect of inhibitor of nuclear factor- 魏 B on the damage of HL7702 hepatocytes induced by hydrogen peroxide, and the effect of L-carnitine on the expression of nuclear factor- 魏 B in HL7702 induced by hydrogen peroxide was detected by Western blot. The nuclear translocation of nuclear factor- 魏 B was determined by cell immunofluorescence staining, and the DNA binding activity of nuclear factor- 魏 B was analyzed by gel migration assay (electrophoretic mobility shift assay,EMSA). Results compared with hydrogen peroxide injury group, the activity of nuclear factor- 魏 B inhibitor group was significantly increased, and the release rate of lactate dehydrogenase was decreased (P0.05, P0.01), and the expression of nuclear factor- 魏 B was increased after hydrogen peroxide injury. L-carnitine preincubation inhibited the nuclear translocation of nuclear factor- 魏 B induced by hydrogen peroxide (P0.01), and L-carnitine also inhibited the increase of the binding activity of nuclear factor- 魏 B DNA induced by hydrogen peroxide. Conclusion L-carnitine inhibits hydrogen peroxide induced hepatocyte injury by inhibiting the activity of nuclear factor-魏 B.
【作者单位】：滨州医学院实验教学管理中心机能学实验室;
【基金】：山东省自然科学基金(No.ZR2010HL068)
【分类号】：R151

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