Protective effect of sevoflurane preconditioning on lung ischemia-reperfusion injury in rats and its influence on TLR4/MyD88/NF-κB signaling pathway
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摘要:
目的 探讨七氟醚预处理对肺缺血-再灌注损伤（IRI）的保护作用及对Toll样受体4（TLR4）/髓样分化因子88（MyD88）/核因子（NF）-κB信号通路的影响。 方法 将40只健康成年SD大鼠随机分为对照组（Sham组）、模型组（LIRI组）、七氟醚预处理组（Sev组）和TLR4抑制剂TAK-242联合七氟醚预处理组（TAK+Sev组），每组各10只。采用苏木素-伊红（HE）染色观察肺组织病理学变化并进行病理损伤评分；采用脱氧核糖核酸末端转移酶介导的dUTP缺口末端标记（TUNEL）法检测肺组织细胞凋亡并计算细胞凋亡率；测定肺组织湿重/干重（W/D）比值以确定肺组织含水量；检测肺组织中氧化应激相关指标水平以及肺组织和血清中炎症因子水平；采用蛋白质印迹法检测肺组织中TLR4/MyD88/NF-κB信号通路相关蛋白的表达水平。 结果 与Sham组比较，LIRI组和Sev组大鼠肺组织病理损伤评分、W/D比值、细胞凋亡率、丙二醛（MDA）水平、炎症因子水平以及TLR4、MyD88、NF-κB p65蛋白相对表达量均升高，超氧化物歧化酶（SOD）水平和NF-κB抑制蛋白α（IκBα）相对表达量均降低（均为P < 0.05）；与LIRI组比较，Sev组和TAK+Sev组大鼠肺组织病理损伤评分、W/D比值、细胞凋亡率、MDA水平、炎症因子水平以及TLR4、MyD88、NF-κB p65蛋白相对表达量均降低，SOD水平和IκBα相对表达量均升高（均为P < 0.05）；与Sev组比较，TAK+Sev组大鼠肺组织病理损伤评分、W/D比值、细胞凋亡率、MDA水平、炎症因子水平以及TLR4、MyD88、NF-κB p65蛋白相对表达量均降低，IκBα相对表达量升高（均为P < 0.05）。 结论 七氟醚预处理能够抑制TLR4/MyD88/NF-κB信号通路的激活，抑制炎症反应和氧化应激，从而有效减轻肺IRI。 Abstract:Objective To evaluate the protective effect of sevoflurane preconditioning on lung ischemia-reperfusion injury (IRI) and its influence on the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor (NF)-κB signaling pathway. Methods Forty healthy adult SD rats were randomly divided into the control group (Sham group), lung IRI model group (LIRI group), sevoflurane group (Sev group) and TLR4 inhibitor TAK-242 combined with sevoflurane group (TAK+Sev group), 10 rats in each group. The pathological changes of lung tissues were observed by hematoxylin-eosin (HE) staining and the pathological injury score was graded. The cell apoptosis of lung tissues was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick- end labeling (TUNEL) and the apoptosis rate was calculated. The wet-to-dry (W/D) ratio of lung tissues was measured to determine the water content of lung tissues. The levels of oxidative stress-related parameters in the lung tissues and inflammatory factors in both the lung tissues and serum were detected. The expression levels of TLR4/MyD88/NF-κB signaling pathway-associated proteins in the lung tissues were determined by Western blot. Results Compared with the Sham group, the pathological injury score, W/D ratio, cell apoptosis rate, malondialdehyde (MDA) level, inflammatory factor level and the relative expression levels of TLR4, MyD88 and NF-κB p65 proteins in the lung tissues were significantly increased, whereas the superoxide dismutase (SOD) level and the relative expression level of NF-κB inhibitory protein α(IκBα) were significantly decreased in the LIRI and Sev groups (all P < 0.05). Compared with the LIRI group, the pathological injury score, W/D ratio, cell apoptosis rate, MDA level, inflammatory factor level and the relative expression levels of TLR4, MyD88 and NF-κB p65 proteins were significantly decreased, whereas the SOD level and the relative expression level of IκBα were significantly increased in the Sev and TAK+Sev groups (all P < 0.05). Compared with the Sev group, the pathological injury score, W/D ratio, cell apoptosis rate, MDA level, inflammatory factor level and the relative expression levels of TLR4, MyD88, NF-κB p65 proteins were significantly decreased, while the relative expression level of IκBα was significantly increased in the TAK+Sev group (all P < 0.05). Conclusions Sevoflurane preconditioning may inhibit the activation of TLR4/MyD88/NF-κB signaling pathway and suppress inflammatory reaction and oxidative stress, thereby effectively mitigating the lung IRI. -
图 1 各组大鼠肺组织的病理学表现（HE，×200）
Figure 1. Pathological findings in lung tissues of rats among each group
图 2 各组大鼠肺组织W/D比值和细胞凋亡率的比较
注：A图为各组大鼠肺组织TUNEL染色结果（×200）; B图为各组大鼠肺组织细胞凋亡率比较; C图为各组大鼠肺组织W/D比值比较。与Sham组比较，aP < 0.05;与LIRI组比较，bP < 0.05;与Sev组比较，cP < 0.05。
Figure 2. Comparison of W/D ratio and cell apoptosis rate in lung tissues of rats among each group
图 3 各组大鼠肺组织MDA和SOD水平的比较
注：与Sham组比较，aP < 0.05;与LIRI组比较，bP < 0.05;与Sev组比较，cP < 0.05。
Figure 3. Comparison of levels of MDA and SOD in lung tissues of rats among each group
图 4 各组大鼠肺组织和血清炎症因子水平的比较
注：与Sham组比较，aP < 0.05;与LIRI组比较，bP < 0.05;与Sev组比较，cP < 0.05。
Figure 4. Comparison of levels of inflammatory factors in lung tissues and serum of rats among each group
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