我们的网站为什么显示成这样？

可能因为您的浏览器不支持样式，您可以更新您的浏览器到最新版本，以获取对此功能的支持，访问下面的网站，获取关于浏览器的信息：

|本期目录/Table of Contents|

脂联素改善低氧诱导的大鼠肺微血管内皮细胞功能障碍及机制研究

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2017年第5期
页码:
529-533
栏目:
基础研究
出版日期:
2017-03-25

文章信息/Info

Title:
Adiponectin improves hypoxia-induced pulmonary microvascular endothelial cell dysfunction and its mechanism
作者:
徐海军1; 4; 张存娟1; 张 娟1; 吴海霞1; 苏 慧2; 张海峰3; 孙 新1
（第四军医大学：1.西京医院儿科，2.西京医院老年病科，3.教学实验中心,陕西 西安 710032；
4.杨凌示范区医院儿科,陕西 杨凌 712100）
Author(s):
XU Hai-jun1; 4;  ZHANG Cun-juan1;  ZHANG Juan1;  WU Hai-xia1;  SU Hui2;  ZHANG Hai-feng3;  SUN Xin1
(1.Department of Pediatrics, 2.Department of Geriatrics, Xijing Hospital, 3.Experiment Teaching Center, Fourth Military Medical University, Xi’an 710032, Shaanxi, China；
4.Department of Pediatrics, Hospital of Yangling Demonstration District, Yangling 712100, Shaanxi, China)
关键词:
肺动脉内皮细胞; 脂联素; 内皮细胞功能障碍
Keywords:
pulmonary artery endothelialcell;  adiponectin;  endothelial dysfunction
分类号:
R589.2
DOI:
-
文献标识码:
A
摘要:
目的 研究重组人球状脂联素（APN）改善低氧诱导的大鼠肺微血管内皮细胞（PMVECs）功能障碍，探讨其潜在的分子机制。方法 取SD雄性大鼠PMVECs原代培养，传至第三代行细胞鉴定；PMVECs分3组，常氧（210 ml/L O2,37℃）组；低氧（20 ml/L O2,37℃）组；低氧（20 ml/L O2,37℃）+APN（1 μg/ml）处理组，分别培养3 h、6 h、9 h和12 h，收集细胞上清测NO浓度；3组PMVEC行RT- PCR测定eNOS基因表达；BCA法测定各组总蛋白含量；行Western blot检测AMPK/p-AMPK、PI3K/p-PI3K、Akt/p-Akt和eNOS/p-eNOS表达。结果 ①鉴定培养细胞为PMVECs；②在相应时间点，与正常组比较，低氧诱导的NO浓度、eNOS mRNA表达水平、总蛋白表达显著下降（P<0.01）；与低氧组对比，低氧+APN组显著增加了低氧诱导的NO浓度、eNOS mRNA表达水平、总蛋白表达（P<0.01）；③3组AMPK、PI3K、Akt和eNOS总蛋白表达水平不变；与正常组比较，低氧诱导的AMPK、PI3K、Akt、eNOS磷酸化表达水平下降（P<0.05）；与低氧组对比，低氧+APN组增加了低氧诱导的AMPK、PI3K、Akt、eNOS磷酸化表达水平（P<0.05）。结论 在细胞水平上，APN能改善低氧条件下诱导的PMVECs功能障碍，促进肺血管具有舒张作用的NO产生，其可能机制可能是伴随着AMPK/PI3K/Akt/eNOS /NO信号通路的激活，APN可能成为潜在治疗低氧性肺动脉高压的辅助药物。
Abstract:
AIM To investigate the effect and molecular mechanism of adiponectin in improving hypoxia-induced dysfunction of rat pulmonary microvascular endothelial cells (PMVECs). METHODS The third passage of primary cultured PMVECs from SD rats were identified by immunohistochemical method and were divided into three groups. The three groups were cultured respectively under normoxia condition (210 ml/L O2, 37℃), hypoxia condition (20 ml/L O2, 37℃) or hypoxia condition (20 ml/L O2, 37℃) with treatment of APN (1 μg/ml). Supernatant and cell lysate were collected at different time intervals at 3 h, 6 h, 9 h and 12 h. NO concentration in supernatant was determined by nitrate reductive enzymatic method, the expression of eNOS-mRNA was observed by RT-PCR, the total protein concentration was detected by the method of BCA, and protein levels of AMPK/p-AMPK, PI3K/p-PI3K,Akt/p-Akt and eNOS/p-eNOS were respectively detected by Western blot from 12 h cells in each group. RESULTS Primary culture of PMVECs was successful. At the same time interval, NO production, mRNA of eNOS and total protein levels in the hypoxia group decreased significantly compared with those in the normoxia group (P<0.01). No production, eNOS mRNA and total protein levels in the hypoxia+ANP group were significantly higher than those in the hypoxia group (P<0.01). There was no significant difference in total protein levels of AMPK, PI3K,Akt and eNOS among the three groups. Phosphorylated AMPK, PI3K,Akt and eNOS decreased significantly（P<0.05） in the hypoxia group compared with those in the normoxia group. Phosphorylated AMPK, PI3K,Akt and eNOS increased significantly（P<0.05） in the hypoxia+ANP group compared with those in the hypoxia group. CONCLUSION APN improves hypoxia-induced dysfunction of PMVECs at cellular levels. It can increase the production of NO possibly through the activation of AMPK/PI3K/Akt/eNOS/NO signaling pathway. APN is a potential ancillary drug for the treatment of hypoxia pulmonary hypertension.

参考文献/References

[1]Gladwin MT.Cardiovascular complications and risk of death in sickle-cell disease[J].Lancet,2016,387(10037):2565-2574.
[2]Farber HW,Loscalzo J.Pulmonary arterial hypertension[J].N Engl J Med,2004,351(16):1655-1665.
[3]Ebrahimi-Mamaeghani M,Mohammadi S,Arefhosseini SR,et al.Adiponectin as a potential biomarker of vascular disease[J].Vasc Health Risk Manag,2015,11:55-70.
[4]Tan KC,Xu A,Chow WS,et al.Hypoadiponectinemia is associated with impaired endothelium-dependent vasodilation[J].J Clin Endocrinol Metab,2004,89(2):765-769.
[5]张存娟,杨 瑞,邢文娟,等.脂联素改善间歇性低氧性肺动脉高压大鼠离体肺动脉舒张功能及其机制[J].心脏杂志,2016,28(3):273-278.
[6]Sakao S,Tatsumi K,Voelkel NF.Endothelial cells and pulmonary arterial hypertension:apoptosis, proliferation,interaction and transdifferentiation[J].Respir Res,2009,10:95.
[7]Ma C,Li Y,Ma J,et al.Key role of 15-lipoxygenase/15-hydroxyeicosatetraenoic acid in pulmonary vascular remodeling and vascular angiogenesis associated with hypoxic pulmonary hypertension[J].Hypertension,2011,58(4):679-688.
[8]Cheng KK,Lam KS,Wang Y,et al.Adiponectin-induced endothelial nitric oxide synthase activation and nitric oxide production are mediated by APPL1 in endothelial cells[J].Diabetes,2007,56(5):1387-1394.
[9]苗兰英,林大勇,白 剑,等.低氧条件下大鼠血管内皮细胞的形态观察[J].生物学杂志,2011,(04):18-20.
[10]Woodfin A,Voisin MB,Nourshargh S.PECAM-1:a multi-functional molecule in inflammation and vascular biology[J].Arterioscler Thromb Vasc Biol,2007,27(12):2514-2523.
[11]Yau JW,Teoh H,Verma S.Endothelial cell control of thrombosis[J].BMC Cardiovasc Disord,2015,15:130.
[12]Rafii S,Butler JM,Ding BS.Angiocrine functions of organ-specific endothelial cells[J].Nature,2016,529(7586):316-325.
[13]Furchgott RF,Zawadzki JV.The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine[J].Nature,1980,288(5789):373-376.
[14]Su JB.Vascular endothelial dysfunction and pharmacological treatment[J].World J Cardiol,2015,7(11):719-741.
[15]Fish JE,Yan MS,Matouk CC,et al.Hypoxic repression of endothelial nitric-oxide synthase transcription is coupled with eviction of promoter histones[J].J Biol Chem,2010,285(2):810-826.
[16]Wang Z,Li AY,Guo QH,et al.Effects of cyclic intermittent hypoxia on ET-1 responsiveness and endothelial dysfunction of pulmonary arteries in rats[J].PLoS One,2013,8(3):e58078.
[17]Giuliano F.New horizons in erectile and endothelial dysfunction research and therapies[J].Int J Impot Res,2008,20(Suppl 2):S2-S8.
[18]Hoyos CM,Melehan KL,Liu PY,et al.Does obstructive sleep apnea cause endothelial dysfunction? A critical review of the literature[J].Sleep Med Rev,2015,20:15-26.
[19]Kadowaki T,Yamauchi T.Adiponectin and adiponectin receptors[J].Endocr Rev,2005,26(3):439-451.
[20]Yamauchi T,Kamon J,Ito Y,et al.Cloning of adiponectin receptors that mediate antidiabetic metabolic effects［J］. Nature, 2003, 423(6941):762-769.
[21]Chen H,Montagnani M,Funahashi T,et al.Adiponectin stimulates production of nitric oxide in vascular endothelial cells[J].J Biol Chem,2003,278(45):45021-45026.

备注/Memo

备注/Memo:
收稿日期：2016-09-26.
基金项目：国家自然基金项目资助（81270330,30700345,31371151和31271219）；陕西省科技厅国际合作课题项目资助（2013KW30-02）；陕西省科学技术研究发展计划项目资助（2013Kkjxx-89）
通讯作者：孙新，副教授，主要从事肺动脉高压研究 Email:sunxin6@fmmu.edu.cn
作者简介：徐海军，硕士生 Emai:navy800628@163.com
更新日期/Last Update: 2017-04-20