乳脂球表皮生长因子8通过调控缺氧诱导生长因子-1α/血管内皮细胞生长因子/Notch信号通路减轻移植肺缺血再灌注损伤  

Protective effects of milk fat globule epidermal growth factor 8 on ischemic reperfusion injury of pulmonary grafts by modulating hypoxia inducible factor-1α/vascular endothelial growth factor/Notch signaling pathway

作　　者：王蓉[1,2] 刘青云 李平[3] 黄振华 尹来波[1] 章爽 高思海[3] Wang Rong;Liu Qingyun;Li Ping;Huang Zhenhua;Yin Laibo;Zhang Shuang;Gao Sihai(Department of Cardiothoracic Surgery,the First Affiliated Hospital,School of Medicine,Shihezi University,Shihezi 832008,China;Department of Medical Record Information Statistics,the First Affiliated Hospital,School of Medicine,Shihezi University,Shihezi 832008,China;Division of Cardiothoracic and Vascular Surgery,Heart-Lung Transplantation Center,Sino-Swiss Heart-Lung Transplantation Institute,Key Laboratory of Organ Transplantation,Ministry of Health,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]石河子大学医学院第一附属医院心胸外科,石河子832008 [2]石河子大学医学院第一附属医院病案信息统计科,石河子832008 [3]华中科技大学同济医学院附属同济医院心脏大血管外科心肺移植中心,武汉430030

出　　处：《中华实验外科杂志》2023年第1期78-82,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金面上项目(81973992)。

摘　　要：目的探讨乳脂球表皮生长因子8(MFG-E8)对移植肺缺血再灌注损伤的保护作用及缺氧诱导生长因子-1α(HIF-1α)/血管内皮细胞生长因子(VEGF)/Notch信号转导通路的影响。方法雄性SD大鼠60只,随机分为移植组、MFG-E8组、MFG-E8+HIF-1α抑制剂组,改良袖套法建立大鼠左肺移植模型,MFG-E8组受体大鼠术前30 min尾静脉注射rhMFG-E820μg/kg,MFG-E8+HIF-1α抑制剂组在MFG-E8组基础上,术前经腹腔注射4 mg/kg HIF-1α抑制剂YC-1。移植组受体大鼠尾静脉注射等量生理盐水。移植肺再灌注2 h后阻断右肺门5 min,行动脉血气分析,并测量移植肺组织测髓过氧化物酶(MPO)活性、肺组织湿/干重量比率(W/D);苏木精-伊红(HE)染色观察移植肺组织病理学变化;原位缺口末端标记法(TUNEL)法检测移植肺组织细胞凋亡指数。蛋白免疫印迹(Western blot)法检测HIF-1α/VEGF/Notch通路蛋白及细胞凋亡蛋白B细胞淋巴瘤/白血病-2(bcl-2)和bcl-2相关X蛋白(bax)表达,酶联免疫吸附法(ELISA)检测肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)表达水平。结果移植肺再灌注2 h后,MFG-E8组动脉血氧分压/吸氧浓度(PaO_(2)/FiO_(2))较移植组明显提升[(307.23±29.17)mmHg比(215.48±20.92)mmHg,t=8.08,P<0.05]。与移植组比较,MFG-E8组肺组织MPO活性和W/D均明显降低[(1.76±0.21)U/g tissue比(2.51±0.46)U/g tissue,(6.03±1.07)mg/dl比(8.41±1.36)mg/dl,t=4.69、4.35,P<0.05]。移植组病理检测可见肺泡壁水肿、肺泡结构破坏、肺泡腔内明显炎性细胞浸润,与移植组比较,MFG-E8组肺泡结构保持完整、肺泡腔和间质内浸润的炎性细胞明显减少,炎性反应显著减轻。与移植组比较,MFG-E8组肺组织细胞凋亡指数(AI)、bax、TNF-α、IL-6蛋白表达显著降低[(11.47±1.23)%比(27.09±2.57)%、(1.34±0.12)比(1.92±0.13)、(250.1±11.76)pg/ml比(392.7±28.14)pg/ml、(134.6±11.25)pg/ml比(251.3±14.33)pg/ml,t=17.34、10.37、14.79、20.26,P<0.05],bcl-2、HIF-1α、VEGF、NotchObjective To investigate the effects of milk fat globule epidermal growth factor 8(MFG-E8)on hypoxia inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF)/Notch signaling pathway in pulmonary grafts with ischemic reperfusion injury.Methods Totally,60 male Sprague Dawley(SD)rats were randomly divided into transplant group,MFG-E8 group,and MFG-E8+HIF-1α inhibitor group.The rat left lung transplantation model was established by a"cuff-like"technique.Rats in MFG-E8 group were given rhMFG-E820μg/kg via the tail vein 30 min before transplantation,and in the MFG-E8+HIF-1α inhibitor group,rats were given rhMFG-E820μg/kg via the tail vein and peritoneal injection of YC-1(HIF-1α inhibitor)4 mg/kg 30 min before transplantation,while the transplant group was injected with the same amount of normal saline.The gas exchange and oxygenation of the grafts were assessed under occlusion of the right pulmonary artery and main bronchus for 5 min after 2-h reperfusion,as well as measurement of Wet/Dry lung weight ratio and myeloperoxidase(MPO)activity.The pathological changes of grafts were observed by hematoxylin-eosin(HE)staining.Apoptosis index of grafts was detected by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)assay.The expression levels of HIF-1α,VEGF,Notch1,HES1,B cell lymphoma/leukemia-2(bcl-2)and bcl-2 associated X protein(bax)were detected by Western blotting,and those of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were detected by enzyme linked immunosorbent assay(ELISA).Results After 2-h reperfusion,compared to the transplant group,the PaO_(2)/FiO_(2) was improved significantly in the MFG-E8 group[(307.23±29.17)vs.(215.48±20.92)mmHg,t=8.08,P<0.05].The activity of MPO and Wet/Dry lung weight ratio in the MFG-E8 group were(1.76±0.21)U/g tissue and(6.03±1.07)mg/dL,respectively,which were significantly lower than those in transplant group[(2.51±0.46)U/g tissue and(8.41±1.36)mg/dL,respectively,t=4.69,4.35,P<0.05].Extensive alveolar hemorrhage,interstitial thick

关 键 词：肺移植 缺血再灌注损伤 缺氧诱导生长因子-1α/血管内皮细胞生长因子/Notch信号通路 细胞凋亡 细胞因子 

分 类 号：R655.3[医药卫生—外科学]