| 摘要: |
| 目的：观察 α7 烟碱型乙酰胆碱受体（α7nAChR）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 α- 银环蛇毒素（α-BGT）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、α-BGT 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（电流 l~2 mA，频率 2/15 Hz，疏密波）；α-BGT 组于模型制备前 30 min 腹腔注射 α7nAChR 拮抗剂 α-BGT（1 μg/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。采用 ELISA 法检测肺组织肿瘤坏死因子 -α（TNF-α）、白介素 -1β（IL-1β）、白介素 -6（IL-6）含量，Western blot 法检测 α7nAChR 蛋白表达水平。结果：模型组、电针组和 α-BGT 组再灌注 4 h 时肺组织 W/D 比值分别为 6.89±1.07、4.56±0.75、8.68±1.49，均明显高于假手术组（2.83±0.43，P<0.05）；肺损伤评分分别为 7.52±1.05、4.16±0.68、9.23±1.57， 均明显高于假手术组（1.19±0.22，P<0.05）；TNF-α 分别为（13.3±2.3）pg/mg、（11.0±1.6）pg/mg、（14.1±3.0）pg/mg， 均明显高于假手术组 [(3.2±0.3) pg/mg，P<0.05] ；IL-1β 分别为（9.2±2.0）pg/mg、（7.4±1.7）pg/mg、（10.5±1.9）pg/mg，均明显高于假手术组 [(2.1±0.5) pg/mg，P<0.05] ；IL-6 分别为（14.2±2.3）pg/mg、（11.7±2.0）pg/mg、（14.8±1.8）pg/mg，均明显高于假手术组 [(4.2±0.7)pg/mg，P<0.05] ；α7nAChR 表达水平分别为（0.55±0.09）、（0.87±0.14）、（0.33±0.08），均明显高于假手术组（0.23±0.04， P<0.05）。与模型组比较，电针组再灌注 4h 时肺组织 W/D 比值、肺损伤评分、肺组织 TNF-α、IL-1β、IL-6 含量均显著降低（P<0.05），电针组 α7nAChR 表达均显著上调（P<0.05）。与电针组比较，α-BGT 组再灌注 4h 时肺组织 W/D 比值、肺损伤评分、肺组织 TNF-α、IL-1β、IL-6 含量升高（P<0.05），α7nAChR 表达下调（P<0.05）。结论：α7nAChR 表达上调，可能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: 烟碱 电刺激疗法 缺血再灌注 肺损伤 兔 |
| DOI：10.3969/j.issn.1007-6948.2019.04.004 |
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| 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 α 7 nAChR in Electroacupuncture-induced Reduction of Lung Injury Induced by Limb Ischemia-reperfusion in Rabbits |
| GONG Li-rong,DOMG Shu-an,KAN Yong-xing,YU Jian-bo |
| Department of Anesthesiology，Tianjin Nankai Hospital Nankai Clinical College of Tianjin Medical University, Tianjin (300270), China |
| Abstract: |
| Objective To observe the effect of α 7 nicotinic acetylcholine receptor (α 7 nAChR) in 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 each): Sham operation group, limb ischemia-reperfusion group, electrical acupoint stimulation group and α-bungarotoxin (α-BGT) group. Limb ischemia-reperfusion was induced by clamping bilateral femoral arteries for 3 h followed by 4 hreperfusion. Bilateral electroacupuncture stimulation of Zusanli and Feishu acupoints was performed (intensity of 1–2 mA, frequency of 2/15 Hz, disperse- dense wave) 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 α-BGT group. At 30 min before the model establishment, speci?c α7nAChR inhibitor α-BGT 1 μg/kg was injected intraperitoneally in α-BGT group. Blood samples were taken from the internal carotid artery at 4 h of reperfusion, and the rabbits were sacrificed. The lungs were removed for microscopic examination and for determination of lung injury scores, wet to dry weight ratio (W/D), contents of TNF-α, interleukin-1beta (IL-1 β) and IL-6, and expression of α 7nAChR by Western blotting. Results The lung W/D ratio after 4 h reperfusion in limb ischemia-reperfusion group, electrical acupoint stimulation group and α-BGT group were 6. 89± 1.07, 4.56±0. 75 and 8. 68± 1. 49, respectively. The lung injury scores in above three groups were 7.52±1.05, 4.16±0.68 and 9.23±1.57, respectively. Both lung W/D ratio and lung injury scores in above three groups were signi?cantly higher than that of Sham operation group. TNF-α concentrations [(13.3± 2.3), ( 11.0± 1.6) and (14.1± 3. 0) pg/mg] in limb ischemia- reperfusion group, electrical acupoint stimulation group and α-BGT group were significantly increased compared with Sham operation group [(3.2 ± 0 . 3 ) pg/mg, P < 0 . 05 ], IL- 1 β concentrations [( 9 . 2 ± 2.0), (7.4 ± 1.7) and (10.5 ± 1 . 9 ) pg/mg] in above three groups were significantly increased compared with Sham operation group [( 2 . 1 ± 0 . 5 ) pg/mg, P<0.05], IL-6 concentrations [(14.2±2.3), (11.7±2.0) and (14.8±1.8) pg/mg] in above three groups were significantly increased compared with Sham operation group [(4. 2 ± 0. 7) pg/mg, P < 0. 05], and expression of α 7 nAChR [( 0 . 55 ± 0.09), (0.87 ± 0 . 14 ) and ( 0 . 33 ± 0 . 08 )] in above three groups were significantly up- regulated compared with Sham operation group [( 0 . 23 ± 0 . 04 ), P < 0 . 05 ]. Compared with limb ischemia- reperfusion group, lung W/D ratio, lung injury scores, contents of TNF-α, IL-1β and IL-6 were signi?cantly decreased, and expression of α7nAChR was signi?cantly up-regulated in electrical acupoint stimulation group. Compared with electrical acupoint stimulation group, lung W/D ratio, lung injury scores, contents of TNF-α, IL-1β and IL-6 were signi?cantly increased, and expression of α7nAChR was signi?cantly down-regulated in α-BGT group. Conclusion Up-regulated expression of α7nAChR is involved in electroacupuncture-induced reduction of lung injury induced by limb ischemia-reperfusion in rabbits. |
| Key words: Nicotinic electrical acupoint stimulation ischemia-reperfusion lung injury rabbits |
