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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(P<0.05,P<0.01);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(P<0.01);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。
Objective To investigate whether L-carnitine inhibits the hepatocyte injury induced by hydrogen peroxide (H 2 O 2) and its relationship with nuclear factor-κB (NF-κB). Methods CCK-8 and lactate dehydrogenase (LDH) release assay was used to determine the effect of NF-κB inhibitor on hydrogen peroxide-induced HL7702 hepatocyte injury. Western blotting was used to detect the effect of L-carnitine on hydrogen peroxide-induced HL7702 The expression of nuclear factor-κB was detected by immunofluorescence staining. The nuclear translocation of nuclear factor-κB was detected by immunofluorescence staining. The DNA binding activity of nuclear factor-κB was analyzed by electrophoretic mobility shift assay (EMSA). Results Compared with hydrogen peroxide injury group, the activity of NF-κB inhibitor group was significantly increased, and the release rate of lactate dehydrogenase was decreased (P <0.05, P <0.01). After H2O2 injury, the levels of nuclear factor-κB Pretreatment with levocarnitine significantly inhibited hydrogen peroxide-induced nuclear factor-κB nuclear translocation (P <0.01), while levocarnitine up-regulated the activity of hydrogen-induced nuclear factor-κB DNA binding Also inhibitory effect. Conclusion L-carnitine inhibits hydrogen peroxide-induced hepatocellular injury by inhibiting nuclear factor-κB activity.