西格列汀通过上调TTF-1/SP-B通路对LPS诱导小鼠急性肺损伤的保护作用  

Sitagliptin exert a protective effect on LPS-induced acute lung injury in mice by upregulating the TTF-1/SP-B pathway

作　　者：王小川 杜发旺 姚宇 李丽[1] 何高燕 王汉超 王勤[2] 熊伟 朱涛[1,3] WANG Xiaochuan;DU Fawang;YAO Yu;LI Li;HE Gaoyan;WANG Hanchao;WANG Qin;XIONG Wei;ZHU Tao(Department of Respiratory Medicine and Critical Care Medicine,and Institute of Respiratory Diseases,Suining Central Hospital,Suining 629000,China;Department of Respiratory Medicine and Critical Care Medicine,and Institute of Respiratory Diseases,Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China;Preclinical Research Center,Suining Central Hospital,Suining 629000,China)

机构地区：[1]遂宁市中心医院呼吸与危重症医学科,遂宁629000 [2]重庆医科大学附属第二医院呼吸与危重症医学科,重庆400010 [3]遂宁市中心医院基础实验室,遂宁629000

出　　处：《西南医科大学学报》2023年第6期495-499,共5页Journal of Southwest Medical University

基　　金：国家自然科学基金青年项目(8180011074);重庆市科卫联合项目(2020FYYX17,2020MSXM112)。

摘　　要：目的 探讨小鼠急性肺损伤(acute lung injury,ALI)状态下DPP4(dipeptidyl peptidase 4 inhibitor,DPP4I)抑制剂西格列汀对肺组织的保护作用及潜在的分子机制。方法 将32只雄性BALB/C小鼠随机分为对照组、DPP4抑制剂西格列汀组(DPP4I组)、内毒素组(LPS组)和LPS+DPP4I组,每组8只。采用气道内注射LPS建立小鼠ALI模型,72 h后获取标本,通过HE染色观察肺组织病理学改变及损伤评分,采用Diff-Quik染色对肺泡灌洗液(alveolar larage fluid,BALF)中炎症细胞分类计数,使用湿/干比(W/D)检测肺水肿程度,使用qPCR和western blotting法分析肺组织中甲状腺转录因子-1(thyroid transcription Factor 1,TTF-1)和表面活性物质相关蛋白-B(surfactant-associated protein-B,SP-B)的mRNA和蛋白表达水平。结果 小鼠气道内注射LPS干预72 h后,肺组织出现明显的病理学改变,肺损伤评分增加,肺水肿指标W/D显著升高,BALF中细胞总数、中性粒细胞数、巨噬细胞数和淋巴细胞数明显增多,肺组织中TTF-1和SP-B的mRNA和蛋白表达水平明显降低,差异均有统计学意义(P <0.05)。对照组与DPP4I组未见明显病理改变。与LPS组相比,LPS+DPP4I组小鼠的肺组织病理改变程度、肺损伤评分、W/D比值、BALF中细胞总数、中性粒细胞数、巨噬细胞数和淋巴细胞数均明显更低,肺组织TTF-1和SP-B的mRNA和蛋白表达水平均明显更高(P <0.05)。结论 在小鼠ALI模型中DPP4抑制剂西格列汀可能通过上调肺组织TTF-1促进SP-B分泌,减轻肺损伤和炎症反应水平,为DPP4I应用于急性呼吸窘迫综合征(acute respiratory distress syndrome,ARDS)的临床治疗奠定了实验基础。Objective This study aims to explore the protective effects of the dipeptidyl peptidase 4 inhibitor,Sitagliptin,on lung tissue in a mouse model of acute lung injury(ALI),as well as its potential molecular mechanisms.Methods Thirty-two male BALB/C mice were randomly divided into 4 groups,Control group,Sitagliptin(DPP4I)group,LPS group,and LPS+DPP4I group,with 8 in each.After 72 hours of interventions,HE staining was used to observe the pathological alterations,subsequently,to evaluate lung injury scores.Wet to dry(W/D)ratio was used to assess pulmonary edema.Inflammatory cells in BALF,including neutrophils,macrophages,lymphocytes,and total cells,were observed and counted using Diff-Quik staining.The mRNA and protein expression of TTF-1 and SPB were measured by RT-PCR and western blotting.Results After 72 hours of interventions,severe lung injury pathological changes,and increased lung injury scores and W/D ratio were observed.Meanwhile,the counts of neutrophils,macrophages,lymphocytes,and total cells in BALF were increased,and the mRNA and protein expression of TTF-1 and SP-B were reduced by LPS.No pathological alterations were observed both in Control group and DPP4I group.Then,compared to LPS group,the pathological changes were attenuated,lung injury scores,W/D ratio,and neutrophils,macrophages,lymphocytes,and total cells in BALF were decreased,and the mRNA and protein expression of TTF-1 and SP-B were up-regulated in LPS+DPP4I group.Conclusions Collectively,our findings indicated that Sitagliptin,a DPP4I,alleviated LPS-induced ALI through up-regulation of TTF-1/SP-B signal pathway in mice.Therefore,these results suggested that dipeptidyl peptidase 4 inhibitor was a potential and promising drug for the treatment of ARDS in clinical practice.

关 键 词：急性肺损伤 DPP4抑制剂 西格列汀 甲状腺转录因子-1 表面活性物质相关蛋白-B 

分 类 号：R-33[医药卫生]