干扰Rho相关卷曲螺旋形成蛋白激酶1表达对急性肺损伤小鼠的保护作用  

Protective Effect of Interfering with Rho-associated Coil Formation Protein Kinase 1 Expression on Mice with Acute Lung Injury

作　　者：李玉芳 杨楠 于小纬 魏志伟 左天昕 邹芳 LI Yufang;YANG Nan;YU Xiaowei;WEI Zhiwei;ZUO Tianxin;ZOU Fang(Respiratory and Critical Care Unit,the First Affiliated Hospital of Hebei North University,Zhangjiakou,Hebei,075000,China)

机构地区：[1]河北北方学院附属第一医院呼吸与危重症监护病房,河北省张家口市075000

出　　处：《医学分子生物学杂志》2025年第2期160-165,共6页Journal of Medical Molecular Biology

基　　金：2023年度河北省医学科学研究课题(No.20231415)。

摘　　要：目的研究干扰Rho相关卷曲螺旋形成蛋白激酶1对脂多糖(lipopolysaccharides,LPS)诱导的急性肺损伤(acute lung injury,ALI)小鼠的影响。方法C57BL/6小鼠随机分为4组:正常对照组、模型组、阴性对照组、ROCK1干扰组,每组10只。腹腔注射5 mg/kg LPS诱导建立急性肺损伤小鼠模型。RTPCR和蛋白质印迹检测Rho相关卷曲螺旋形成蛋白激酶1(Rho-associated coiled coil formation protein kinase 1,ROCK1)表达水平,肺功能检测系统检测干扰ROCK1对小鼠气道阻力、静息通气量和肺容积的影响,HE染色观察干扰ROCK1对肺组织结构的影响,Masson染色观察干扰ROCK1对肺纤维化程度的影响,蛋白质印迹检测α-SMA、TGF-β1和FN蛋白水平,TUNEL检测干扰ROCK1对肺组织细胞凋亡的影响,ELISA检测iNOS、IL-6和TNF-α炎性因子的含量水平。结果与正常对照组比较,模型组小鼠ROCK1高表达,肺损伤严重。与模型组比较,ROCK1干扰组小鼠ROCK1低表达(P<0.05),气道阻力、静息通气量和肺容积均显著升高(P<0.05),肺组织病理损伤和纤维化程度均得到改善,α-SMA、TGF-β1、FN蛋白水平显著降低(P<0.05),肺凋亡细胞数显著减少(P<0.05),外周血中iNOS、IL-6和TNF-α含量水平显著降低(P<0.05)。结论干扰ROCK1表达能够改善LPS诱导的急性肺损伤小鼠肺功能、肺纤维化、细胞凋亡和炎症。Objective To investigate the effect of interference with Rho-associated coil forming protein kinase 1 on lung injury induced by lipopolysaccharide(LPS)in mice.Methods C57BL/6 mice were divided into 4 groups:healthy control group,model group,negative control group and ROCK1 interference group,10 mice each group.Acute lung injury was induced with LPS(5 mg/kg,i.p.).The expression level of ROCK1 was detected by RT-PCR and Western blotting.The pulmonary function detection system was used to detect the airway resistance,resting ventilation and lung volume in mice.HE staining was used to observe the pathological damage of lung tissues,and Masson staining was used to observe the pulmonary fibrosis.Western blotting was used to detect the protein expression levels ofα-SMA,TGF-β1 and FN.TUNEL assay was used to detect the lung cell apoptosis.The levels of iNOS,IL-6 and TNF-αwere detected by ELISA.Results ROCK1 was highly expressed in the model group and the lung injury was more serious when compared with that in the healthy control group.ROCK1 interference group had lower expression level of ROCK1(P<0.05),and increased airway resistance,resting ventilation and lung volume when compared with the model group(P<0.05).The pathological injury and degree of fibrosis in lung tissues were improved in the ROCK1 interference group,and the expression levels ofα-SMA,TGF-β1 and FN proteins were significantly decreased(P<0.05).The number of apoptotic cells in lung tissues was significantly decreased(P<0.05),and the levels of iNOS,IL-6 and TNF-αin peripheral blood were significantly decreased in the ROCK1 interference group when compared with those in the model group(P<0.05).Conclusion Interfering ROCK1 expression can improve lung function,pulmonary fibrosis,apoptosis and inflammation in LPS-induced acute lung injury mice.

关 键 词：ROCK1 脂多糖 急性肺损伤 

分 类 号：R563.9[医药卫生—呼吸系统]