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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Western blot检测NF-κB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-κB活化(P<0.05)。(4)抗氧化剂NAC抑制缺氧诱导的NF-κB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-κB活化,从而抑制缺氧诱导的大鼠神经元凋亡。
Objective To investigate the effect of naloxone on neuronal damage induced by hypoxia in rats and its possible mechanism. Methods Rat neurons were cultured in vitro and divided into normal control group, hypoxia induction group and hypoxia induction + naloxone intervention group. The neuronal damage was detected by flow cytometry. The changes of intracellular reactive oxygen species (ROS) were detected by fluorescent probe 2,7-dichlorodihydrofluorescein acetoacetate (DCFDA). The levels of superoxide dismutase SOD) activity; Western blot detection of NF-κB activation. Results (1) Naloxone inhibited neuronal apoptosis induced by hypoxia, the apoptosis of neurons in hypoxia group was significantly increased 3.54 times that of the control group and 1.35 times that of the naloxone intervention group (All P <0.05). (2) Naloxone inhibited hypoxia-induced reactive oxygen species (ROS) production. Hypoxia induced ROS production by 2.66 folds of the control group and 1.24 folds of the naloxone intervention group (all P <0.05) . (3) Naloxone inhibited hypoxia-induced activation of NF-κB (P <0.05). (4) Antioxidant NAC can inhibit hypoxia-induced activation of NF-κB, the inhibition rate was 49% (P <0.05). Conclusion Naloxone can inhibit the hypoxia-induced ROS generation, and then inhibit the activation of NF-κB, thereby inhibiting hypoxia-induced neuronal apoptosis.