纳洛酮对缺氧诱导大鼠神经元损伤的抑制作用及其机制

目的探讨纳洛酮对缺氧诱导大鼠神经元损伤的作用及其可能的机制。方法体外培养大鼠神经元，将其分为正常对照组、缺氧诱导组、缺氧诱导＋纳洛酮干预组。应用流式细胞术测定大鼠神经元损伤；应用荧光探针2，7-二氯二氢荧光素乙酰乙酸（DCFDA）检测细胞内活性氧的变化；应用比色法检测超氧化物歧化酶（SOD）的活性；应用Westernblot检测NF—KB的活化。结果（1）纳洛酮抑制缺氧诱导的大鼠神经元凋亡，缺氧组大鼠神经元细胞凋亡明显增加，为对照组的3．54倍，纳洛酮干预组为对照组的1．35倍（均P〈0．05）。（2）纳洛酮抑制缺氧诱导的活性氧生成，缺氧诱导大鼠神经元活性氧生成，为对照组的2．66倍，纳洛酮干预组为对照组的1．24倍（均P〈0．05）。（3）纳洛酮抑制缺氧诱导的NF—KB活化（P〈0．05）。（4）抗氧化剂NAC抑制缺氧诱导的NF—KB活化，抑制率达49％（P〈0．05）。结论纳洛酮抑制缺氧诱导的活性氧生成，进而抑制NF—KB活化，从而抑制缺氧诱导的大鼠神经元凋亡。

Study on naloxone blocked hypoxia-induced rat neuron injury

Objective To investigate the effects of naloxone on hypoxia-induced rat neuron injury. Methods Rat neuron were cultured and divided into control group, hypoxia-induced group, naloxone pretreated group. Neuron apoptosis was detected by using flow cytometry. Fluorescent probe, 2, 7- dichloro-fluorescein diacetate （DCFDA）, was used to examine intracellular reactive oxygen species （ROS） generation. Superoxide dismutase （SOD） activity was accessed spectrophotometrieally by using SOD assay kit. NF-KB activation was detected by western blotting. Results （1） Naloxone inhibited hypoxia-induced rat neuron apoptosis. Hypoxic rats neuronal apoptosis significantly increased 3.54 times than that of the control group（ all P 〈 0.05 ）. （2） Naloxone inhibited hypoxia-induced ROS generation. Hypoxia-induced neuron reactive oxygen species generation, was 2.66 times than that of the control group, the naloxone pretreatment group was 1.24 times than that of the control group （ all P 〈 0.05 ）. （ 3 ） Naloxone inhibited hypoxia-induced NF-KB activation （P 〈 0.05 ）. （4） The antioxidant, NAC, inhibited hypoxia-induced NF-KB activation with inhibition rate of 49% （ P 〈 0.05 ）. Conclusion Naloxone can prevent hypoxia-induced rat neuronal apoptosis by inhibiting hypoxia-induced ROS generation in rat neuron and further blocking NF-KB activation.