恶性脑肿瘤缺失蛋白1在脓毒症致急性呼吸窘迫综合征大鼠中的表达和作用  

Expression and role of deleted in malignant brain tumor protein 1 in acute respiratory distress syndrome rats induced by sepsis

作　　者：葛晨 张一凡 贾红炜 罗钰泉 祖一文 杨志伟 贾丽静 王景梅[3] 杜全胜[1] Ge Chen;Zhang Yifan;Jia Hongwei;Luo Yuquan;Zu Yiwen;Yang Zhiwei;Jia Lijing;Wang Jingmei;Du Quansheng(Department of Intensive Care Unit,Hebei General Hospital,Shijiazhuang 050000,Hebei,China;Department of Intensive Care Unit,Shijiazhuang People's Hospital,Shijiazhuang 050000,Hebei,China;Department of Intensive Care Unit,Handan Central Hospital,Handan 056000,Hebei,China)

机构地区：[1]河北省人民医院重症医学科,石家庄050000 [2]石家庄市人民医院重症医学科,河北石家庄050000 [3]邯郸市中心医院重症医学科,河北邯郸056000

出　　处：《中华危重病急救医学》2023年第2期152-157,共6页Chinese Critical Care Medicine

基　　金：河北省自然科学基金(H2020307040);中国博士后科学基金(2020M670024ZX);河北省博士后科研项目(B2019003039);河北省医学科学研究重点计划项目(20180192);河北省政府资助临床医学人才培养计划项目(2020003)。

摘　　要：目的观察恶性脑肿瘤缺失蛋白1(DMBT1)在脓毒症致急性呼吸窘迫综合征(ARDS)大鼠中的表达及其与ARDS相关生物标志物的关系。方法取48只健康雄性Wistar大鼠,按随机数字表法分为假手术(Sham)组和ARDS模型组,两组再根据术后6、12、24 h分为3个亚组,每个亚组8只。Sham组单纯开腹暴露盲肠;ARDS模型组采用盲肠结扎穿孔术(CLP)制备脓毒症致ARDS模型。分别于术后6、12、24 h观察大鼠的一般表现;取大鼠腹主动脉血,采用酶联免疫吸附试验(ELISA)检测血清DMBT1、肺表面活性物质相关蛋白D(SP-D)、血管内皮生长因子(VEGF)、白细胞介素(IL-6、IL-10)水平。取肺组织,测定肺湿/干质量(W/D)比值;苏木素-伊红(HE)染色后,光镜下观察肺组织病理学改变,同时进行肺组织损伤评分;采用蛋白质免疫印迹试验(Western blotting)检测DMBT1蛋白在肺组织中的表达。采用Pearson相关法分析血清DMBT1与SP-D、VEGF、IL-6、IL-10及肺组织损伤评分的相关性。结果ARDS模型组大鼠术后出现明显病态表现,光镜下可见肺泡结构破坏、炎症细胞浸润、肺泡出血等,术后12 h起肺组织损伤评分较Sham组明显升高(分:3.35±0.13比1.16±0.07,P<0.05),且肺W/D比值较Sham组明显增加(5.36±0.44比4.38±0.35,P<0.05),存在肺水肿,提示脓毒症致ARDS模型制备成功。ELISA及Western blotting检测结果显示,ARDS模型组大鼠血清DMBT1、SP-D、VEGF、IL-6水平均随时间延长呈逐渐升高趋势,IL-10呈先升高后下降趋势;与Sham组比较,ARDS模型组血清及肺组织DMBT1的蛋白表达于术后6 h起即明显升高〔血清(ng/L):231.96±19.17比187.44±10.19,肺组织(DMBT1/β-actin):2.05±0.19比0.93±0.25,均P<0.05〕,而血清SP-D、VEGF、IL-6、IL-10水平则于术后12 h起明显升高〔SP-D(ng/L):73.35±8.05比43.28±5.77,VEGF(ng/L):89.85±8.47比43.19±5.11,IL-6(ng/L):36.01±2.48比17.49±1.77,IL-10(ng/L):84.55±8.41比39.83±5.02,均P<0.05〕。Pearson相关分析显示,ARDS模�Objective To observe the expression of deleted in malignant brain tumor protein 1(DMBT1)in rat acute respiratory distress syndrome(ARDS)model induced by sepsis and its relationship with ARDS related biomarkers.Methods Forty-eight healthy male rats were randomly divided into sham operation group(Sham group)and ARDS model group,and the rats in each group were further divided into three subgroups at 6,12 and 24 hours after operation,with 8 rats in each subgroup.The rats in the Sham group were exposed to the cecum only,and sepsis induced ARDS model was reproduced by cecal ligation and puncture(CLP)in the ARDS model group.The general performance was observed at 6,12,24 hours after operation.Abdominal aortic blood of rats was collected,and the levels of DMBT1,surfactant-associated protein D(SP-D),vascular endothelial growth factor(VEGF),interleukins(IL-6,IL-10)in serum were determined by enzyme-linked immunosorbent assay(ELISA).The lung tissues were collected,and the lung wet/dry weight(W/D)ratio was determined.The lung tissue pathological changes were observed under light microscope after hematoxylin-eosin(HE)staining,and the lung tissue injury score was evaluated.The expression of DMBT1 protein in lung tissue was determined by Western blotting.The relationship between the serum DMBT1 and SP-D,VEGF,IL-6,IL-10,lung tissue injury score were analyzed by Pearson correlation analysis.Results Rats in the ARDS model group showed obvious pathological manifestations after operation.The alveolar structure destruction,inflammatory cell infiltration,and alveolar hemorrhage were observed under microscope.Compared with the Sham group,the lung tissue injury score and the lung W/D ratio at 12 hours after operation in the ARDS model group were significantly increased(lung tissue injury score:3.35±0.13 vs.1.16±0.07,lung W/D ratio:5.36±0.44 vs.4.38±0.35,both P<0.05),and pulmonary edema was present,which suggested that the ARDS model caused by CLP was successfully reproduced.The results of ELISA and Western blotting showed that the l

关 键 词：恶性脑肿瘤缺失蛋白1 急性呼吸窘迫综合征 脓毒症 生物标志物 肺泡上皮细胞损伤 毛细血管通透性 炎症反应 

分 类 号：R459.7[医药卫生—急诊医学] R563.8[医药卫生—治疗学]