南方医科大学学报

南方医科大学学报 ›› 2018, Vol. 38 ›› Issue (09): 1061-.doi: 10.12122/j.issn.1673-4254.2018.09.06

• • 上一篇    下一篇

低强度脉冲超声预处理通过激活胆碱能抗炎通路抑制HMGB1表达减轻肺缺血再灌注损伤

曲良超，严金秀，蒋章颉，宋志平，罗佛全，彭清华   

  • 出版日期:2018-09-20 发布日期:2018-09-20

Low-intensity pulsed ultrasound pretreatment inhibits HMGB1 expression and attenuates lung ischemia-reperfusion injury in rats via the cholinergic anti-inflammatory pathway

  • Online:2018-09-20 Published:2018-09-20

HTML

PDF

1353

摘要： 目的通过建立大鼠肺缺血再灌注损伤（IRI）模型，进行低强度脉冲超声（LIPUS）预处理治疗，探讨IRI后高迁移率蛋白1 （HMGB1）在肺组织的表达和LIPUS 预处理的保护作用。方法雄性SD大鼠32 只，体质量250~300 g，随机分为4 组，每组8 只。对照组（A组），开胸游离左肺门，未行阻断；缺血再灌注组（B组），阻断左肺门45 min后再灌注180 min；预处理组（C组）低 强度超声波治疗仪超声定位辐照30 min，然后同B组处理；预处理加α7-烟碱型胆碱能受体（α7nAChR）拮抗剂组（D组），LIPUS 预处理前30 min腹腔注射α7nAChR拮抗剂甲基牛扁亭2 mg/Kg,然后同预处理组处理。测量肺组织湿干重比值（W/D）和肺通 透指数（LPI），大鼠肺组织病理学观察及评分，ELISA法测定肺组织IL1和IL6的浓度，免疫荧光和Western blot检测HMGB1蛋 白表达。结果与A组比较，其他三组IR后肺组织W/D值（5.75±0.47）和LPI（2.77±0.18）明显升高（P<0.05），病理学评分（13.31± 2.82）明显升高（P<0.05），肺组织IL1（69.13±9.11）和IL6（62.77±8.14）水平明显升高（P<0.05），免疫荧光检测平均IOD值（0.046± 0.019）和Western blot检测显示HMGB1表达明显增加（P<0.05）。给于LIPUS预处理后，C组IR后肺组织W/D值（5.07±0.28）和 LPI（1.85±0.17）比B组明显降低（P<0.05），病理学评分（8.13±1.76）比B组明显降低（P<0.05），同时肺组织IL1 和IL6 水平以及 HMGB1表达也明显降低，给于α7nAChR拮抗剂后该作用明显被抑制。结论LIPUS预处理能够减轻IRI后肺损伤，其机制可能 是通过激活依赖α7nAChR的胆碱能抗炎通路，从而降低肺组织HMGB1的表达。

Abstract: Objective To observe the effects of low-intensity pulsed ultrasound (LIPUS) pretreatment on pulmonary expression of high mobility group box-1 (HMGB1) in a rat model of lung ischemia-reperfusion (IR). Methods Thirty-two male Sprague- Dawley rats weighing 250-300 g were randomly divided (n=8) into sham-operated group, lung IR group, LIPUS pretreatment group and pretreatment with α7-nicotinic cholinergic receptor (α7nAChR) antagonist group. In the sham-operated group, the left pulmonary hilum was dissociated without occlusion; in the other 3 groups, the left pulmonary hilum was occluded for 45 min followed by reperfusion for 180 min; LIPUS pretreatment for 30 min and intraperitoneal injection of methyllycaconitine (2 mg/kg), an α7nAChR antagonist, were administered before the operation. The wet/dry weight ratio (W/D) and pulmonary permeability index (LPI) of the lung tissue were measured, and the lung histopathology was observed and scored. The contents of interleukin-1 (IL- 1) and IL-6 in the lung tissues were measured using ELISA, and the pulmonary expression of HMGB1 protein was detected using immunofluorescence assay and Western blotting. Results Compared with those in the sham-operated group, the W/D of the lung tissue, LPI, pathological scores, IL-1 and IL-6 contents in the lung tissue, and pulmonary HMGB1 expression all significantly increased in the other 3 groups (P<0.05). LIPUS preconditioning significantly lowered the W/D values, LPI, pathological score, IL-1 and IL-6 contents and HMGB1 expression in the lung tissues following lung IR, and these effects were significantly inhibited by administration of methyllycaconitine. Conclusions LIPUS preconditioning can reduce lung IR injury possibly by activating α7nAChR-dependent cholinergic anti-inflammatory pathway to reduce lung tissue HMGB1 expression.