维生素D类似物通过激活VDR/GPX4通路改善小鼠呼吸机相关性肺损伤  

Vitamin D analogues activate vitamin D receptor/glutathione peroxidase 4 pathway to improve ventilator-induced lung injury in mice

作　　者：谢秋雯 邵容格 谢永果[1] 潘灵辉 秦科[1] 杜学柯[1] Xie Qiuwen;Shao Rongge;Xie Yongguo;Pan Linghui;Qin Ke;Du Xueke(Department of Anesthesiology,the Second Affiliated Hospital of Guangxi Medical University,Nanning 530007,Guangxi Zhuang Autonomous Region,China;Guangxi Clinical Research Center for Anesthesiology,Guangxi Engineering Research Center for Tissue&Organ Injury and Repair Medicine,Guangxi Key Laboratory for Basic Science and Prevention of Perioperative Organ Disfunction,Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学第二附属医院麻醉科,南宁530007 [2]广西麻醉学临床医学研究中心,广西壮族自治区组织器官损伤修复医学工程研究中心,广西围术期器官功能损伤与防治基础研究重点实验室,南宁530021

出　　处：《中华危重病急救医学》2022年第4期383-387,共5页Chinese Critical Care Medicine

基　　金：广西自然科学基金(2018GXNSFAA138007);广西医疗卫生适宜技术开发与推广应用项目(S2018104);广西医疗卫生重点学科建设项目(2020-9-17)。

摘　　要：目的研究维生素D类似物paricalcitol激活维生素D受体/谷胱甘肽过氧化物酶4(VDR/GPX4)通路在呼吸机相关性肺损伤(VILI)中的作用。方法将24只雄性C57BL/6J小鼠随机分为对照组、大潮气量(HVT)致VILI模型组(HVT组)、paricalcitol对照组(P组)和paricalcitol预处理组(P+HVT组),每组6只。给予小鼠气管插管,按40 mL/kg的潮气量通气制备VILI模型;对照组仅气管插管,不予通气。P+HVT组小鼠于制模前1周腹腔注射paricalcitol 0.2μg/kg,每日1次;P组仅在实验前1周腹腔注射paricalcitol 0.2μg/kg,每日1次。通气4 h后处死小鼠取肺组织,通过肺湿/干质量(W/D)比值、苏木素-伊红(HE)染色和Masson染色评价肺损伤情况;采用蛋白质免疫印迹试验(Western blotting)和免疫组化法检测VDR、GPX4的表达;采用微量法检测丙二醛(MDA)和还原型谷胱甘肽(GSH)的含量。结果HVT通气4 h后小鼠出现明显肺损伤,表现为:与对照组相比,肺损伤评分和肺W/D比值明显升高〔肺损伤评分(分):0.430±0.035比0.097±0.025,肺W/D比值:4.860±0.337比3.653±0.332,均P<0.05〕,胶原纤维沉积明显增加,且肺组织MDA含量明显升高(nmol/g:212.420±8.757比97.073±5.308,P<0.05),GSH含量及VDR、GPX4的蛋白表达和免疫组化评分(IRS)明显降低〔GSH(μg/g):44.229±1.690比70.840±0.781;VDR蛋白(VDR/GAPDH):0.518±0.029比0.762±0.081,GPX4蛋白(GPX4/GAPDH):0.452±0.032比0.649±0.034;IRS评分(分):VDR为4.168±0.408比10.167±0.408,GPX4为4.333±1.033比10.333±0.516;均P<0.05〕。给予paricalcitol激活VDR后,与HVT组相比,P+HVT组小鼠肺损伤明显改善,表现为肺损伤评分和肺W/D比值明显降低〔肺损伤评分(分):0.220±0.036比0.430±0.035,肺W/D比值:4.015±0.074比4.860±0.337,均P<0.05〕,胶原纤维沉积减少,且肺组织MDA含量明显下降(nmol/g:123.840±8.082比212.420±8.757,P<0.05),GSH含量及VDR、GPX4的蛋白表达和IRS评分明显上调〔GSH(μg/g):63.094±0.992比44.229±1.690;VDR蛋白(VDR/GAPDH):0.713±0.056�Objective To investigate the role of vitamin D analogue paricalcitol in activating vitamin D receptor/glutathione peroxidase 4(VDR/GPX4)pathway in ventilator-induced lung injury(VILI).Methods Twenty-four male C57BL/6J mice were randomly divided into control group,high tidal volume(HVT)induced VILI model group(HVT group),paricalcitol control group(P group),and paricalcitol pretreatment group(P+HVT group),with 6 mice in each group.The mice were endotracheal intubated and ventilated at 40 mL/kg tidal volume to prepare VILI model,while those in the control group were intubated without ventilation.The mice in the P+HVT group were intraperitoneally injected with paricalcitol 0.2μg/kg once a day 1 week before modeling,while those in the P group were intraperitoneally injected paricalcitol 0.2μg/kg once a day for 1 week before the experiment.After ventilation for 4 hours,the mice were sacrificed for lung tissue collection.Lung injury was evaluated by wet/dry(W/D)ratio,hematoxylin-eosin(HE)staining and Masson staining.The expressions of VDR and GPX4 were determined by Western blotting and immunohistochemistry.Malondialdehyde(MDA)and glutathione(GSH)contents were determined by micro method.Results After HVT for 4 hours,compared with the control group,lung injury score and W/D ratio were significantly higher(lung injury score:0.430±0.035 vs.0.097±0.025,lung W/D ratio:4.860±0.337 vs.3.653±0.332,both P<0.05),collagen fiber deposition was significantly increased,the content of MDA in lung tissue was significantly increased(nmol/g:212.420±8.757 vs.97.073±5.308,P<0.05),GSH content and the protein expressions and immunoreactive score(IRS)of VDR and GPX4 were significantly decreased[GSH(μg/g):44.229±1.690 vs.70.840±0.781;VDR protein(VDR/GAPDH):0.518±0.029 vs.0.762±0.081,GPX4 protein(GPX4/GAPDH):0.452±0.032 vs.0.649±0.034;IRS score:VDR was 4.168±0.408 vs.10.167±0.408,GPX4 was 4.333±1.033 vs.10.333±0.516;all P<0.05],which meant that the mice in HVT group showed obvious lung injury.After VDR was activated by parica

关 键 词：维生素D受体 谷胱甘肽过氧化物酶4 维生素D类似物 氧化还原稳态 肺损伤 

分 类 号：R563[医药卫生—呼吸系统]