川芎嗪通过Rac1/LIMK1通路减轻脂多糖诱导的小鼠急性肺损伤  

Tetramethylpyrazine attenuates acute lung injury induced by lipopoly⁃saccharide via Rac1/LIMK1 signaling pathway in mice

作　　者：闵思敏 刘小碣 丁渡山 张小楠 陶伟婷 李言[1,3] MIN Si-min;LIU Xiao-jie;DING Du-shan;ZHANG Xiao-nan;TAO Wei-ting;LI Yan(Department of Pathophysiology,Bengbu Medical College,Bengbu 233030,China;The First Affiliated Hospital of Beng-bu Medical College,Bengbu 233030,China;Changzhou University,Changzhou 213000,China)

机构地区：[1]蚌埠医学院病理生理学教研室,安徽蚌埠233000 [2]蚌埠医学院第一附属医院,安徽蚌埠233000 [3]常州大学,江苏常州213000

出　　处：《中国病理生理杂志》2022年第8期1407-1415,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81673791);安徽省高校研究生科学研究项目(No.YJS20210528)。

摘　　要：目的:探讨川芎嗪(TMP)对脂多糖(LPS)诱导的小鼠急性肺损伤(ALI)的预防作用及对Ras相关C3肉毒素底物1(Rac1)/LIM激酶1(LIMK1)信号通路的影响。方法:将C57BL/6小鼠随机分为6组:对照组、模型组(LPS组)、地塞米松(Dex)组及TMP(40、80和120 mg/kg)组。模型组小鼠第1次采用LPS腹腔注射,16 h后第2次采用气管滴注LPS诱导小鼠ALI模型;TMP组小鼠在LPS第1次腹腔注射前30 min及第2次气管滴注LPS后30 min时腹腔注射TMP;Dex组小鼠与TMP组同一时间腹腔注射3 mg/kgDex。LPS气管滴注24 h后检测小鼠外周血氧饱和度(SpO_(2));取小鼠肺组织,计算肺组织湿干重比值(W/D);ELISA法测定肺组织中白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α);HE染色观察肺组织病理学变化;Westernblot检测肺组织中Rac1和LIMK1蛋白水平;支气管肺泡灌洗液中检测白细胞含量及蛋白浓度。结果:与对照组比较,模型组小鼠SpO_(2)降低(P<0.05),W/D升高(P<0.01),肺组织中IL-1β和TNF-α水平均升高(P<0.01);肺泡壁增厚,大量红细胞渗出,可见片状出血;支气管肺泡灌洗液中白细胞总数及蛋白浓度增高(P<0.01);Rac1和p-LIMK1蛋白表达水平显著升高(P<0.01)。与模型组相比,Dex组和TMP组W/D降低(P<0.01),肺组织IL-1β和TNF-α水平降低(P<0.01),肺泡间质水肿及出血减轻,红细胞渗出减少;支气管肺泡灌洗液中白细胞总数及蛋白浓度降低(P<0.01)。与模型组相比,Dex组及低、中剂量TMP组Rac1蛋白水平降低(P<0.01),p-LIMK1在Dex组及TMP组也显著降低(P<0.05,P<0.01),LIMK1在各组的表达无显著差异。结论:TMP可通过抑制Rac1/LIMK1信号通路降低毛细血管通透性,从而减轻LPS诱导的小鼠ALI。AIM:To investigate the effect of tetramethylpyrazine(TMP)on lipopolysaccharide(LPS)-in-duced acute lung injury(ALI)in mice.METHODS:C57BL/6 mice were randomly divided into control group,model group,dexamethasone(Dex)group and TMP(40,80 and 120 mg/kg)groups.A mouse model of ALI was reproduced by intraperitoneal injection of LPS firstly,and intratracheal instillation of LPS 16 h later.The mice in TMP groups were in-jected with TMP intraperitoneally 30 min before the first injection of LPS and 30 min after the tracheal drip of LPS.The mice in Dex group were intraperitoneally injected with 3 mg/kg of Dex.After 24 h of LPS treatment,peripheral blood oxy-gen saturation(SpO_(2))was measured and the lung tissue was harvest for all mice.The ratio of lung wet-to-dry weight(W/D)was calculated.Concentrations of interleukin-1β(IL-1β)and tumor necrosis factorα(TNF-α)in lung tissues were measured by ELISA.HE staining was used to observe the pathological changes of lung tissues.Western blot was used to detect the protein levels of Ras-related C3 botulinum toxin substrate 1(Rac1)and Lim kinase 1(LIMK1)in lung tissues.The total number of leukocytes and protein concentration in bronchoalveolar lavage fluid were detected.RESULTS:Com-pared with control group,SpO_(2) was decreased(P<0.05),W/D was increased(P<0.01),and the levels of IL-1βand TNF-αin lung tissues of model group was increased(P<0.01).Increased alveolar wall thickness,erythrocyte effusion,and increased total leukocyte count and protein concentration in bronchoalveolar lavage fluid were observed in the lung tis-sue of model group.The protein levels of Rac1 and p-LIMK1 were increased in model group(P<0.01).Compared with model group,W/D,the levels of IL-1βand TNF-α,the edema and bleeding of alveolar stroma in lung tissues,and eryth-rocyte effusion were decreased in both Dex group and TMP groups.Compared with model group,the expression level of Rac1 protein in Dex group,low-dose TMP group and middle-dose TMP group were decreased(P<0.01).p-LIMK1 was al-so decreased sign

关 键 词：川芎嗪 急性肺损伤 Rac1/LIMK1信号通路 

分 类 号：R363.2[医药卫生—病理学] R563[医药卫生—基础医学]