川芎嗪对LPS致心肌细胞损伤的影响及机制研究

目的探讨川芎嗪对脂多糖(LPS)诱导的大鼠心肌细胞损伤及NF-κB核移位的影响。方法采用原代培养SD乳鼠心肌细胞,经终浓度分别为40、80、120μmol·L-1川芎嗪预处理后,用10 mg·L-1LPS处理6 h,处理完成后检测培养液乳酸脱氢酶(LDH)活性,四唑盐(MTT)比色法检测心肌细胞存活率,流式细胞法检测ROS生成,试剂盒检测细胞内丙二醛(MDA)含量及细胞内抗氧化酶(SOD、GSH-Px)活性、Western blot法检测核蛋白中NF-κB p65的变化情况。结果不同剂量川芎嗪(40、80、120μmol·L-1)预处理3 h后可明显降低LDH活性,增加细胞存活率,降低ROS生成,减少MDA含量,升高SOD、GSH-Px活性,抑制NF-κB p65在细胞核中的表达,且呈剂量依赖性。结论川芎嗪可抑制LPS所致的心肌损伤,其机制与减少脂质过氧化、增强抗氧化酶系、降低ROS生成、抑制NF-κB p65核移位有关。

Effects of TMPZ on LPS-induced cardiac cell damage and involved mechanism

Aim To explore the role of tetramethylpyrazine(TMPZ) in lipopolysaccharide(LPS)-induced cardiac cell damage and the nuclear translocation of nuclear factor kappaB(NF-κB).Methods Cardiac cells isolated from primary neonatal SD rat were pretreated with 40,80,120 μmol·L-1 TMPZ respectively,and then treated with 10 mg·L-1 LPS for 6 hours.After treatment,the activity of lactic acid dehydrogenase(LDH) was determined by auto-biochemistry analysator,cells viability was analyzed by methyl thiazolyl tetrazolium(MTT),generation of reactive oxygen species(ROS) were measured by flow cytometry,the content of malondialdehyde(MDA),and the activity of superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) were analyzed by colorimetric method,the level of NF-κB p65 protein in nuclear was detected by western blotting.Results 3 hours pretreatment with different doses of TMPZ(40,80,120 μmol·L-1) significantly increased cardiac cells viability,decreased LDH activity,ROS production and MAD content.SOD and GSH-Px activities were improved,and the nuclear expression of NF-κB p65 was inhibited in a dose-dependent manner.Conclusion TMPZ can alleviate LPS-induced cardiac cell damage by suppressing lipid peroxidation,enhancing antioxidant enzymes,decreasing ROS production and inhibiting nuclear translocation of NF-κB.