摘要
目的评价胆碱能抗炎通路(CAP)在右美托咪定(DEX)抑制大鼠机械通气肺损伤(VILI)及VILI时NLRP3炎症小体激活中的作用。方法将48只SD大鼠随机分为自主呼吸对照组(C组)、大潮气量组(H组)、DEX组、α-7烟碱型乙酰胆碱受体拮抗剂α-银环蛇毒素(α-BGT)组,每组12只。4组大鼠均行气管切开插管术。C组大鼠保持自主呼吸,H组、DEX组、α-BGT组大鼠均以20 mL/kg潮气量行机械通气,DEX组和α-BGT组大鼠于机械通气前1 h经颈内静脉输注DEX 5μg/(kg·h),α-BGT组于输注DEX前15 min经腹腔注射α-BGT 1μg/kg。4 h后麻醉处死大鼠,收集血清、肺组织标本、支气管肺泡灌洗液(BALF)。光镜下观察肺组织病理学改变,测定肺损伤评分及肺组织湿/干质量比(W/D);应用逆转录-聚合酶链反应(RT-PCR)及蛋白质免疫印迹试验(Western Blot)分别检测肺组织中NLRP3、凋亡相关斑点样蛋白(ASC)、天冬氨酸半胱氨酸蛋白酶-1(caspase-1)的mRNA及蛋白表达;应用酶联免疫吸附试验(ELISA)测定BALF中总蛋白含量以及血清、BALF中白细胞介素(IL)-1β、IL-18的含量。结果与C组比较,H组、DEX组、α-BGT组大鼠肺损伤评分、肺组织W/D和BALF中总蛋白含量均升高(F=3.130~8.266,t=2.270~11.900,P<0.05),肺组织内NLRP3、ASC及caspase-1的mRNA及蛋白表达均上调(F=2.384~45.256,t=2.195~10.992,P<0.05),血清和BALF中IL-1β、IL-18的含量均升高(F=2.053~35.522,t=3.109~8.242,P<0.05);与H组比较,DEX组上述各指标均降低(t=2.313~5.487,P<0.05);与DEX组比较,α-BGT组上述各指标均升高(t=2.195~5.050,P<0.05)。结论 DEX通过抑制NLRP3炎症小体的激活减轻大鼠VILI,其作用机制与CAP有关。
Objective To investigate the role of the cholinergic anti-inflammatory pathway(CAP) in the inhibition of ventilator-induced lung injury(VILI) by dexmedetomidine(DEX) and the activation of NACHT-LRR-PYD-containing proteins 3(NLRP3) inflammasome in rats. Methods A total of 48 Sprague-Dawley rats were randomly divided into spontaneous brea-thing control group(group C), large tidal volume group(group H), DEX group(group DEX), and α7 nAChR antagonist α-BGT group(group α-BGT), with 12 rats in each group. Tracheostomy and intubation were performed for all groups. The rats in group C maintained spontaneous breathing, and those in groups H, DEX, and α-BGT were given mechanical ventilation with a tidal vo-lume of 20 mL/kg;the rats in groups DEX and α-BGT received the infusion of DEX 5 μg/(kg·h) via the internal jugular vein at 1 h before mechanical ventilation, and those in group α-BGT were given intraperitoneal injection of α-BGT 1 μg/kg at 15 min before DEX infusion. The rats were anesthetized and sacrificed 4 h later to collect the samples of blood serum, lung tissue, bronchoalveolar lavage fluid(BALF). Lung histopathological changes were observed under a light microscope, and lung injury score and wet/dry(W/D) ratio of lung tissue were measured. RT-PCR and Western blot were used to measure the mRNA and protein expression of NLRP3, apoptosis-associated speck-like protein(ASC), and caspase-1 in lung tissue, and ELISA was used to measure the level of total protein in BALF and the levels of interleukin-1β(IL-1β) and interleukin-18(IL-18) in serum and BALF. Results Compared with group C, groups H, DEX, and α-BGT had significant increases in lung injury score, W/D ratio of lung tissue, and content of total protein in BALF(F=3.130-8.266,t=2.270-11.900,P<0.05), significant increases in the mRNA and protein expression of NLRP3, ASC, and caspase-1 in lung tissue(F=2.384-45.256,t=2.195-10.992,P<0.05), and significant increases in the levels of IL-1β and IL-18 in serum and BALF(F=2.053-35.522,t=3.109-8.242,P<0.05). C
作者
张本旺
侯欣位
于春锐
孙立新
ZHANG Benwang;HOU Xinwei;YU Chunrui;SUN Lixin(Department of Anesthesiology,Qingdao Municipal Hospital Affiliated to Qingdao University,Qingdao 266071,China)
出处
《青岛大学学报（医学版）》
CAS
2020年第6期704-709,共6页
Journal of Qingdao University(Medical Sciences)
基金
青岛市民生科技计划项目(15-9-2-72-nsh)。
关键词
右美托咪啶
呼吸机相关性肺损伤
胆碱能抗炎通路
炎性体
大鼠
dexmedetomidin
ventilator-induced lung injury
cholinergic anti-inflammatory pathway
inflammasome
rats
