内皮抑制素抑制RhoA/ROCK信号通路减轻脂多糖诱导的小鼠急性肺损伤实验研究  

Endostatin attenuates lipopolysaccharide-induced acute lung injury in mice by inhibiting RhoA/ROCK signaling pathway

作　　者：岳渊 张涛[1] 柳朝阳[2] 张鹏霞[1] 肖漓[3] 张娟[3] YUE Yuan;ZHANG Tao;LIU Zhaoyang;ZHANG Pengxia;XIAO Li;ZHANG Juan(School of Basic Medicine,Jiamusi University,Key Laboratory of Microecology-immunomodulatory Network and Related Diseases,Jiamusi 154007,Heilongjiang Province,China;The Fourth Department of Neurology,the First Affiliated Hospital of Jiamusi University,Jiamusi 154003,Heilongjiang Province,China;Institute of Respiratory and Critical Care Medicine,the Eighth Medical Center,Chinese PLA General Hospital,Beijing Key Laboratory of Organ Transplantation and Immune Regulation,Beijing 100091,China)

机构地区：[1]佳木斯大学基础医学院,微生态-免疫调节网络与相关疾病重点实验室,黑龙江佳木斯154007 [2]佳木斯大学第一附属医院神经四科,黑龙江佳木斯154003 [3]解放军总医院第八医学中心呼吸与危重症医学部研究所,北京市器官移植与免疫调节重点实验室,北京100091

出　　处：《解放军医学院学报》2023年第9期1001-1009,共9页Academic Journal of Chinese PLA Medical School

基　　金：首都卫生发展科研专项(首发2022-2-5092);黑龙江省卫生健康委科研课题(20210202040058)。

摘　　要：背景急性肺损伤(acute lung injury,ALI)是一种由多种诱因引发的急性肺部炎症性疾病,目前无有效防治手段。内皮抑制素(endostatin,ES)可特异性抑制血管内皮细胞的增殖和迁移,广泛应用于肿瘤治疗中,但其在急性肺损伤中的作用尚不明确。目的探讨内皮抑制素对脂多糖(lipopolysaccharide,LPS)诱导的小鼠急性肺损伤的影响及可能机制。方法将36只雄性C57BL/6小鼠随机分为6组,分别为对照组、LPS组(模型组)、ES干预组A(LPS+1 mg/kg ES 6 h)、ES干预组B(LPS+1 mg/kg ES 12 h)、ES干预组C(LPS+5 mg/kg ES 6 h)、ES干预组D(LPS+5 mg/kg ES 12 h),每组6只。LPS组:将LPS溶液(15 mg/kg)通过肺部液体定量雾化器装置递送至小鼠肺,作用24 h,建立小鼠急性肺损伤模型;对照组:在相同时间点以同样的方式递送等体积0.9%氯化钠注射液,作用24 h;ES干预组:LPS刺激小鼠24 h后,腹腔注射不同剂量(1 mg/kg或5 mg/kg)的ES注射液,分别作用6 h与12 h。观察各组小鼠肺组织形态学改变;测定肺系数变化;行苏木素-伊红染色病理学检查;酶联免疫吸附实验检测小鼠血清中血管内皮生长因子水平;流式细胞术检测小鼠血清与肺泡灌洗液中TNF-α和IL-6水平;脱氧核糖核苷酸末端转移酶介导的缺口末端标记法检测小鼠肺组织细胞凋亡;免疫印记实验检测小鼠肺组织中Ras同源基因家族成员A(Ras homolog gene family member A,RhoA)和Rho相关卷曲螺旋形成蛋白激酶1(Rho-associated coiled-coil-forming protien kinase 1,ROCK1)的表达。结果与对照组比较,LPS组肺系数增大(P<0.01),肺损伤评分升高(P<0.01),肺组织凋亡细胞数增多(P<0.01),血清血管内皮生长因子水平升高(P<0.05),血清与BALF中TNF-α和IL-6水平升高(P<0.05),肺组织RhoA与ROCK1蛋白表达增加(P<0.05)。与LPS组比较,ES干预组D肺部炎性渗出浸润减少,肺间质水肿及肺泡结构破坏减轻,肺损伤评分降低(P<0.01),肺系数减小(P<0.01),肺组织凋亡细胞减少Background Acute lung injury(ALI)is an acute pneumonia disease caused by a variety of incentives.At present,there is no effective prevention and treatment method.Endostatin(ES)can specifically inhibit the proliferation and migration of vascular endothelial cells,which is widely used in tumor therapy,but its role in acute lung injury remains unclear.Objective To investigate the effect of ES on lipopolysaccharide(LPS)-induced acute lung injury in mice and its possible mechanism.Methods A total of 36male C57BL/6 mice were randomly divided into control group,LPS group(model group),ES intervention group A(LPS+1 mg/kgES for 6 h),ES intervention group B(LPS+1 mg/kg ES for 12 h),ES intervention group C(LPS+5 mg/kg ES for 6 h),and ESintervention group D(LPS+5 mg/kg ES for 12 h),with 6 mice in each group.In the LPS group,LPS solution was delivered to thelungs of mice according to the dose of 15 mg/kg through the lung liquid quantitative atomization device,and the acute lung injurymodels of mice were established after LPS injection for 24 hours.In the control group,the same volume of saline was delivered inthe same way at the same time point for 24 hours.In the ES intervention groups,mice were intraperitoneally injected with differentdoses of ES(1 mg/kg or 5 mg/kg)for 6 h or 12 h at 24 h after LPS stimulation.The morphological changes of lung tissues wereobserved.The changes of lung coefficient were measured.Hematoxylin-eosin(HE)staining was used for pathological examination.Serum levels of vascular endothelial growth factor(VEGF)were measured by enzyme-linked immunosorbent assay(ELISA).Thecontents of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in serum and bronchoalveolar lavage fluid(BALF)weredetected by flow cytometry.Apoptosis of lung cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotinnick end labeling assay(TUNEL).The expression levels of Ras homolog gene family member A(RhoA)and Rho-associated coiledcoil-forming protein kinase 1(ROCK1)in lung tissues of mice in each group were d

关 键 词：内皮抑制素 急性肺损伤 RhoA/ROCK信号通路 细胞因子 炎症反应 

分 类 号：R459.7[医药卫生—急诊医学]