二氧化硫对大鼠肢体缺血再灌注致急性肺损伤p38丝裂原活化蛋白激酶信号转导通路的影响  

Effects of sulfur dioxide on p38 mitogen-activated protein kinase signal transduction pathway in acute lung injury induced by limb ischemia reperfusion in rats

作　　者：赵彦瑞[1] 单磊[1] 王东[1] 刘洋[1] 周君琳[1] Zhao Yanrui;Shan Lei;Wang Dong;Liu Yang;Zhou Junlin(Department of Orthopedics,Beijing Chaoyang Hospital,Capital Medical University,Beijing 100020,China)

机构地区：[1]首都医科大学附属北京朝阳医院骨科,北京100020

出　　处：《中国医学前沿杂志（电子版）》2021年第4期63-68,共6页Chinese Journal of the Frontiers of Medical Science(Electronic Version)

基　　金：国家自然科学基金(81070050);北京市自然科学基金(7152061)。

摘　　要：目的探讨外源性二氧化硫(sulfur dioxide,SO_(2))抗肢体缺血再灌注(ischemia reperfusion,IR)致急性肺损伤的作用机制及对p38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)信号转导通路的影响。方法通过绑扎大鼠双侧后腿根部4 h然后放松2 h制作IR致急性肺损伤模型。将32只大鼠随机分为对照组(无肢体缺血操作,8只)、IR组(制备肢体缺血模型,8只)、IR+SO_(2)组(制备肢体缺血模型,并于再灌注前20 min给予SO_(2)供体Na_(2)SO_(3)/NaHSO_(3)腹腔注射,8只)和IR+SO_(2)+SB203580组(在IR+SO_(2)组操作基础上于再灌注前1 h给予SB203580静脉注射,8只)。试验完成后处死大鼠,收集其肺组织及外周血,应用原位末端转移酶标记法、酶联免疫吸附测定、蛋白质印迹法等方法检测细胞凋亡情况、细胞因子表达、p38 MAPK蛋白表达及其磷酸化情况等。结果IR组大鼠肺组织损伤加重,凋亡细胞数量增多,凋亡指数、血浆和肺组织中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素-1β(interleukin-1β,IL-1β)、白介素-10(interleukin-10,IL-10)及白介素-6(interleukin-6,IL-6)表达均显著高于对照组(均P<0.05);IR+SO_(2)组大鼠肺组织学形态改变明显改善,凋亡指数、血浆和肺组织中TNF-α、IL-1β及IL-6表达均显著低于IR组(均P<0.05),血浆和肺组织中IL-10表达均显著高于IR组(均P<0.05),磷酸化p38 MAPK表达上调;IR+SO_(2)+SB203580组大鼠肺组织损伤程度较IR+SO_(2)组加重,凋亡指数、血浆和肺组织中TNF-α、IL-1β及IL-6表达均显著高于IR+SO_(2)组(均P<0.05),而血浆和肺组织中IL-10表达均显著低于IR+SO_(2)组(均P<0.05),磷酸化p38 MAPK表达减少。结论外源性SO_(2)对肢体IR致急性肺损伤具有保护作用,且p38 MAPK信号通路参与介导了SO_(2)抗肢体IR损伤的分子机制,它的活化可以减轻IR损伤。Objective To explore the mechanism of exogenous sulfur dioxide(SO_(2))against limb ischemia reperfusion(IR)induced acute lung injury(ALI)and its effect on the signal transduction pathway of p38 mitogen-activated protein kinase(p38 MAPK).Method ALI model induced by IR was established by binding the roots of both hind legs of rats for 4 h and then relaxing for 2 h.Thirty-two rats were randomly divided into control group(no limb ischemia operation,n=8),IR group(limb ischemia model,n=8),IR+SO_(2) group(limb ischemia model was established,and the SO_(2) donor Na_(2)SO_(3)/NaHSO_(3) was intraperitoneally injected 20 min before reperfusion,n=8)and IR+SO_(2)+SB203580 group(on the basis of operation in IR+SO_(2) group,intravenous injection of SB203580 was given 1 h before reperfusion,n=8).After completion of the experiment,the rats were sacrificed,and lung tissues and peripheral blood were collected.Cell apoptosis,cytokine expression,p38 MAPK protein expression and phosphorylation were detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay,enzyme linked immunosorbent assay and Western blotting.Result In IR group,the lung tissue damage was aggravated,the number of apoptotic cells increased,and the apoptotic index,the expression of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-10(IL-10)and interleukin-6(IL-6)in plasma and lung tissue were significantly higher than those in control group(all P<0.05).The morphological changes of the lung tissue of rats in IR+SO_(2) group were significantly improved.Apoptosis index,the expression of TNF-α,IL-1βand IL-6 in plasma and lung tissue were significantly lower than those in IR group(all P<0.05),expression of IL-10 in plasma and lung tissue were significantly higher than that in IR group(all P<0.05),and the expression of phosphorylated p38 MAPK was up-regulated.The degree of lung tissue damage in IR+SO_(2)+SB203580 group was more severe than that in IR+SO_(2) group.Apoptosis index,expression of TNF-α,IL-1βand IL-6 in pl

关 键 词：P38丝裂原活化蛋白激酶 二氧化硫 缺血再灌注 急性肺损伤 

分 类 号：R-332[医药卫生] R563