| 摘要: |
| 目的：观察核因子E2相关因子2（Nrf2）/血红素氧合酶-1（HO-1）信号通路在电针减轻肢体缺血再灌注诱发兔肺损伤中的作用。方法：40只健康雄性新西兰大白兔，采用随机数字表法分为假手术组、模型组、电针组和全反式维甲酸组（n=10）。采用夹闭股动脉3 h、再灌注4 h的方法制备兔肢体缺血再灌注模型；电针组、全反式维甲酸组于模型制备前1~4 d（30 min/次，1次/d）及模型制备过程中电针足三里穴和肺俞穴，采用疏密波2/15 Hz，强度以兔出现轻微肌颤为宜；全反式维甲酸组于模型制备前30 min腹腔注射Nrf2抑制剂全反式维甲酸7 mg/kg。再灌注4 h时采集颈动脉血样，随后处死兔取肺组织，观察病理学结果并行肺损伤评分，计算肺湿/干重（W/D）比值；测定肺组织MDA含量及SOD活性，采用Western blotting法检测肺组织Nrf2总蛋白、Nrf2核蛋白及HO-l蛋白表达水平。结果：模型组、电针组和全反式维甲酸组再灌注4 h时肺损伤评分分别为5.43±0.82、4.03±0.56、6.21±0.75，均明显高于假手术组（0.92±0.21，P<0.05）；肺组织W/D比值分别为6.42±1.00、4.45±0.68、7.08±1.12，均明显高于假手术组（2.04±0.29，P<0.05）；MDA含量分别为（4.82±0.51）nmol/mg、（3.56±0.45）nmol/mg、（5.18±0.47）nmol/mg，均明显高于假手术组
（2.10±0.23）nmol/mg，P<0.05；SOD活性分别为（50.6±6.3）U/mg、（63.1±7.7）U/mg、（47.2±5.6）U/mg，均明显低于假手术组 （72.6±6.3）U/mg，P<0.05；Nrf2总蛋白、Nrf2核蛋白及HO-l蛋白表达水平均明显高于假手术组（P<0.05）。与模型组比较，电针组再灌注4 h时肺损伤评分、肺组织W/D比值、MDA含量均明显降低（P<0.05），SOD活性、Nrf2总蛋白、核蛋白及HO-l蛋白表达水平均明显升高（P<0.05）。与电针组比较，全反式维甲酸组再灌注4 h时肺损伤评分、肺组织W/D比值、MDA含量均明显升高（P<0.05），SOD活性、Nrf2总蛋白、核蛋白及HO-l蛋白表达水平均明显降低（P<0.05）。
结论：Nrf2/HO-1信号通路激活参与了电针减轻肢体缺血再灌注诱发兔肺损伤的过程。 |
| 关键词: NF-E2相关因子2 血红素氧合酶-1 电刺激疗法 缺血再灌注 肺损伤 |
| DOI： |
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| 基金项目:基金项目：天津市滨海新区卫生计生委科技联合攻关项目(2015BWKL003) |
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| Effect of Nrf2/HO-1 Pathway on Electroacupuncture-induced Reduction of Lung Injury Induced by Limb Ischemia-reperfusion in Rabbits |
| WANG De-qiang, YU Jian-bo, KAN Yong-xing, et al. |
| Department of Anesthesiology, Dagang Hospital of Tianjin Binhai New Area, Tianjin (300270), China |
| Abstract: |
| Objective To observe the effect of nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway on electroacupuncture-induced reduction of lung injury induced by limb ischemia- reperfusion in rabbits. Methods Forty New Zealand white rabbits were randomly divided into 4 groups (n=10), such as Sham operation group, limb ischemia-reperfusion group, electrical acupoint stimulation group and all-trans retinoic acid group. Limb ischemia-reperfusion was induced by clamping bilateral femoral arteries for 3 h followed by 4 h reperfusion. Bilateral electroacupuncture stimulation of Zusanli and Feishu acupoints was performed with disperse-dense wave 2/15 Hz for 30 min once daily for 4 d before establishment of the model and during the course of establishment of the model in electrical acupoint stimulation group and all-trans retinoic acid group. At 30 min before establishment of the model, Nrf2 inhibitor all-trans retinoic acid (7 mg/kg) was injected intraperitoneally in all-trans retinoic acid group. The rabbits were sacrificed at 4 h of reperfusion and the lungs were removed for microscopic examination and determination of lung injury scores, wet to dry weight ratio (W/D), malondialdehyde (MDA) contents, superoxide dismutase (SOD) activities, and expression of Nrf2 total protein, nucleoprotein and HO-1 protein by Western blotting. Results The lung W/D ratio after 4 h reperfusion in limb ischemia-reperfusion group, electrical acupoint stimulation group and all-trans retinoic acid group were 6.42±1.00, 4.45±0.68 and 7.08±1.12, respectively. The lung injury scores at 4 h in above three groups were 5.43±0.82, 4.03±0.56 and 6.21±0.75, respectively. Both lung W/D ratio and lung injury scores in above three groups were significantly higher than those of Sham operation group. MDA contents [(4.82±0.51) nmol/mg, (3.56±0.45) nmol/mg and (5.18±0.47) nmol/mg] in limb ischemia-reperfusion group, electrical acupoint stimulation group and all-trans retinoic acid group were significantly increased compared with Sham operation group [(2.10±0.23) nmol/mg, P<0.05]. The SOD activities [(50.6±6.3) U/mg, (63.1±7.7) U/mg and (47.2±5.6) U/mg] in limb ischemia-reperfusion group, electrical acupoint stimulation group and all-trans retinoic acid group were significantly decreased compared with Sham operation group [(72.6±6.3) U/mg, P<0.05]. The expression of Nrf2 total protein, nucleoprotein and HO-1 protein in above three groups were significantly up-regulated compared with Sham operation group (P<0.05). Compared with limb ischemia-reperfusion group, lung W/D ratio, lung injury scores and MDA contents were significantly decreased, while SOD activities, expression of Nrf2 total protein, nucleoprotein and HO-1 protein were significantly increased in electrical acupoint stimulation group. Compared with electrical acupoint stimulation group, the lung W/D ratio, lung injury scores and MDA contents were significantly increased, while SOD activities, expression of Nrf2 total protein, nucleoprotein and HO-1 protein were significantly decreased in all-trans retinoic acid group. Conclusion Activation of Nrf2/HO-1 signaling pathway is involved in electroacupuncture-induced reduction of lung injury induced by limb ischemia-reperfusion in rabbits. |
| Key words: NF-E2 related factor 2 heme oxygenase-1 electrical acupoint stimulation ischemia-reperfusion lung injury |
