浏览全部资源
扫码关注微信
高原肺水肿大鼠肺组织基因表达谱分析
Gene expression profile of lung tissues in rats with high altitude pulmonary edema
- 陆军军医大学学报 2024年46卷第11期 页码：1235-1243
- 作者机构：
1.400038 重庆，陆军军医大学(第三军医大学)高原军事医学系：高原特需药品与器材研究室
2.400038 重庆，陆军军医大学(第三军医大学)高原军事医学系：高原生理学与病理学教研室
3.400038 重庆，陆军军医大学(第三军医大学)高原军事医学系：极端环境医学教育部重点实验室(培育)
- 作者简介：
[高文祥, E-mail: winneygaocn@aliyun.comGAO Wenxiang, E-mail: winneygaocn@aliyun.com
[陈德伟, E-mail: cdw528913@163.comCHEN Dewei, E-mail: cdw528913@163.com
- 基金信息：国家自然科学基金青年科学基金(82101968)
DOI：10.16016/j.2097-0927.202307015
中图分类号： R394-33;R541.63;R594.3收稿：2023-07-03，
修回：2023-09-25，
纸质出版：2024-06-15
- 稿件说明：
移动端阅览
徐刚, 吴刚, 孙滨达, 等. 高原肺水肿大鼠肺组织基因表达谱分析[J]. 陆军军医大学学报, 2024,46(11):1235-1243.
Gang XU, Gang WU, Binda SUN, et al. Gene expression profile of lung tissues in rats with high altitude pulmonary edema[J]. Journal of Army Medical University, 2024, 46(11): 1235-1243.
徐刚, 吴刚, 孙滨达, 等. 高原肺水肿大鼠肺组织基因表达谱分析[J]. 陆军军医大学学报, 2024,46(11):1235-1243. DOI： 10.16016/j.2097-0927.202307015.
Gang XU, Gang WU, Binda SUN, et al. Gene expression profile of lung tissues in rats with high altitude pulmonary edema[J]. Journal of Army Medical University, 2024, 46(11): 1235-1243. DOI： 10.16016/j.2097-0927.202307015.
摘要
目的
2
利用基因芯片技术分析高原肺水肿(high altitude pulmonary edema，HAPE)大鼠模型肺组织差异表达基因，为深入探讨HAPE发生的分子机制提供新线索。
方法
2
将8周龄健康雄性SD大鼠[体质量(200±20) g
]
按随机数字表法分为常氧对照(NC)组、脂多糖(Lipopolysaccharide，LPS)组、低压低氧(Hypoxia)组和低压低氧复合低剂量LPS(HL)组。LPS组和HL组按照每100 g体质量尾静脉注射0.1 mL浓度为0.05%的LPS溶液，NC组和Hypoxia组给予等量生理盐水；Hypoxia组和HL组进行模拟海拔5 000 m低压低氧处理6 h，NC组和LPS组于舱外同时饲养。检测肺组织的湿/干质量比(wet/dry mass ratio，WDR)、支气管肺泡灌洗液(bronchoalveolar lavage fluid，BALF)蛋白含量，HE染色观察肺组织病理改变。提取肺组织总RNA，采用Affymetrix基因芯片检测mRNA表达谱，采用Metascape在线基因注释和分析系统(
http://metascape.org
http://metascape.org
)对差异表达基因进行基因本体论(gene ontology，GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes，KEGG)信号通路聚类分析。
结果
2
HL组大鼠肺组织表现出明显的充血、水肿、肺泡间隔增宽。与NC组比较，HL组大鼠肺WDR显著增加(
P
＜0.01)，BALF蛋白含量显著升高(
P
＜0.05)。基因表达检测发现，相对NC组，Hypoxia组有79个基因表达上调，59个基因表达下调，LPS组473个基因表达上调，695个基因表达下调，HL组669个基因表达上调，1 253个基因表达下调。GO和KEGG信号通路聚类分析显示：HL组差异表达上调基因主要富集于细胞因子介导的信号通路、对IL-1的反应、炎症反应调节等生物学过程，以及细胞因子-细胞因子受体相互作用、TNF、NF-κB、IL-17、补体和凝血级联反应等信号通路；表达下调的基因主要富集到细胞外基质组织、内皮细胞迁移调控、细胞-基质黏附等生物学过程，以及黏着斑、Wnt、cGMP-PKG、PI3K-Akt、Rap1等信号通路。HL组大鼠肺组织中
NF-κB
、
TNF-α
、
IL
-1
β
以及
IL
-6的mRNA表达均显著上调(
P
＜0.01)。
结论
2
低氧复合低剂量LPS可稳定复制大鼠HAPE模型，低氧可显著增强LPS诱导的炎症、免疫反应，显著增强炎症介质的表达，促进HAPE的发生发展。
Abstract
Objective
2
To analyze the differential expressed genes (DEGs) in the lung tissues of rat model of high altitude pulmonary edema (HAPE) by using microarray analysis in order to provide new clues for molecular mechanism of HAPE.
Methods
2
Healthy male SD rats (8 weeks old
weighing 200±20 g) were randomized into normoxia control (NC) group
lipopolysaccharide (LPS) group
hypoxia group and hypoxia +low-dose LPS (HL) group. The rats of the LPS gr
oup and HL group were injected with 0.1 mL 0.05% LPS per 100 g body weight
and those of the NC group and the hypoxia group were administered with an equivalent volume of normal saline. The rats of the hypoxia group and the HL group were housed in a hypobaric chamber simulating an altitude of 5 000 m
and those of the NC group and the LPS group were raised simultaneously outside of the chamber. The wet/dry mass ratio (WDR) of lung tissue and total protein content in bronchoalveolar lavage fluid (BALF) were measured
and the histopathological changes of lung tissue was observed using HE staining. The total RNA was extracted from the lung tissues
and the mRNA expression profile was obtained with Affymetrix microarray followed by Gene Ontology (GO) analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis with Metascape (
http://metascape.org
http://metascape.org
).
Results
2
The rats of the HL group showed significant congestion
edema
and widened alveolar septa. Compared with the NC group
the HL group had significantly increased lung WDR (
P
<
0.01) and total protein content in BALF (
P
<
0.05). Gene expression analysis revealed that there were 79 genes up-regulated and 59 genes down-regulated in the hypoxia group
473 genes up-regulated and 695 genes down-regulated in the LPS group
and especially
669 genes up-regulated and 1 253 genes down-regulated in the HL group. GO and KEGG pathway analyses revealed that the upregulated genes in the HL group were mainly enriched in biological processes
such as cytokine mediated signaling pathways
response to IL-1
regulation of inflammatory response
as well as signaling pathways
including cytokine-cytokine receptor interactions
TNF
NF-κB
IL-17
complement and coagulation cascades
etc
. The down-regulated genes were mainly enriched in biological processes
such as extracellular matrix organization
regulation of endo
thelial cell migration
cell substrate adhesion
as well as signaling pathways
such as focal adhesion
Wnt
cGMP-PKG
PI3K-Akt
Rap1
etc
. The mRNA expression of
NF-κB
TNF-α
IL
-1
β
and
IL
-6 was significantly up-regulated in the lung tissue of the HL group (
P
<
0.01).
Conclusion
2
Hypoxia+low-dose LPS is an effective procedure to establish a reliable model for HAPE in rats. Hypoxia can significantly aggravate LPS-induced inflammation and immune response
enhance the expression of inflammatory mediators
and thus promote the pathogenesis of HAPE.
关键词
Keywords
references
HACKETT P H, RENNIE D, LEVINE H D. The incidence, importance, and prophylaxis of acute mountain sickness[J]. Lancet, 1976, 2(7996): 1149-1155. DOI: 10.1016/s0140-6736(76)91677-9.
石建国, 季华庆, 陈想贵, 等. 高原肺水肿患者血流动力学变化研究[J]. 中国全科医学, 2021, 24(17): 2192-2197. DOI: 10.12114/j.issn.1007-9572.2021.00.473.
SHI J G, JI H Q, CHEN X G, et al . Hemodynamic changes in patients with high-altitude pulmonary edema[J ] . Chin Gen Pract, 2021, 24(17): 2192-2197. DOI: 10.12114/j.issn.1007-9572.2021.00.473.
林法迎, 赵汝频, 杨琼慧. 高原肺水肿256例临床分析[J]. 临床内科杂志, 2006, 23(2): 113-114. DOI: 10.3969/j.issn.1001-9057.2006.02.013.
LIN F Y, ZHAO R P, YANG Q H. Retrospective clinical analysis on high altitude pulmonary edema[J]. J Clin Intern Med, 2006, 23(2): 113-114. DOI: 10.3969/j.issn.1001-9057.2006.02.013.
BERG J T. Ginkgo biloba extract prevents high altitude pulmonary edema in rats[J ] . High Alt Med Biol, 2004, 5(4): 429-434. DOI: 10.1089/ham.2004.5.429.
陈有, 高钰琪, 李素芝, 等. 高原肺水肿动物模型的初步研究[J]. 第三军医大学学报, 2006, 28(11): 1176-1177. DOI: 10.3321/j.issn:1000-5404.2006.11.014.
CHEN Y, GAO Y Q, LI S Z, et al . Primary establishment of HAPE animal model[J ] . Acta Acad Med Mil Tertiae. 2006, 28(11): 1176-1 177. DOI: 10.3321/j.issn:1000-5404.2006.11.014.
吴刚, 高文祥, 高钰琪. HIF-1: 高原肺水肿防治的新靶点[J]. 中国高原医学与生物学杂志, 2020, 41(1): 8-10, 16. DOI: 10.13452/j.cnki.jqmc.2020.01.003.
WU G, GAO W X, GAO Y Q. HIF-1: a new target for prevention and treatment of high altitude pulmonary edema[J]. Chin High Alt Med Bio. 2020, 41(1): 8-10, 16. DOI: 10.13452/j.cnki.jqmc.2020.01.003.
WU G, XU G, CHEN D W, et al . Hypoxia exacerbates inflammatory acute lung injury via the Toll-like receptor 4 signaling pathway[J ] . Front Immunol, 2018, 9: 1667. DOI: 10.3389/fimmu.2018.01667..
ZHOU Y Y, ZHOU B, PACHE L, et al . Metascape provides a biologist-oriented resource for the analysis of systems-level datasets[J ] . Nat Commun, 2019, 10(1): 1523. DOI: 10.1038/s41467-019-09234-6.
IMRAY C, BOOTH A, WRIGHT A, et al . Acute altitude illnesses[J ] . BMJ, 2011, 343: d4943. DOI: 10.1136/bmj.d4943.
CIESIELSKA A, MATYJEK M, KWIATKOWSKA K. TLR4 and CD14 trafficking and its influence on LPS-induced pro-inflammatory signaling[J]. Cell Mol Life Sci, 2021, 78(4): 1233-1261. DOI: 10.1007/s00018-020-03656-y.
PENA E, SIQUES P, BRITO J, et al . Nox2 upregulation and p38α MAPK activation in right ventricular hypertrophy of rats exposed to long-term chronic intermittent hypobaric hypoxia[J ] . Int J Mol Sci, 2020, 21(22): 8576. DOI: 10.3390/ijms21228576.
SAMANTA D, SEMENZA G L. Maintenance of redox homeostasis by hypoxia-inducible factors[J]. Redox Biol, 2017, 13: 331-335. DOI: 10.1016/j.redox.2017.05.022.
PHAM K, PARIKH K, HEINRICH E C. Hypoxia and inflammation: insights from high-altitude physiology[J]. Front Physiol, 2021, 12: 676782. DOI: 10.3389/fphys.2021.676782.
LI J, YUAN W, JIANG S, et al . Prolyl-4-hydroxylase domain protein 2 controls NF-κB/p65 transactivation and enhances the catabolic effects of inflammatory cytokines on cells of the nucleus pulposus[J ] . J Biol Chem, 2015, 290(11): 7195-7207. DOI: 10.1074/jbc.M114.611483.
ARYA A, SETHY N K, SINGH S K, et al . Cerium oxide nanoparticles protect rodent lungs from hypobaric hypoxia-induced oxidative stress and inflammation[J ] . Int J Nanomedicine, 2013, 8: 4507-4520. DOI : 10.2147/IJN.S53032.
SHARMA M, SINGH S B, SARKAR S. Genome wide expression analysis suggests perturbation of vascular homeostasis during high altitude pulmonary edema[J]. PLoS One, 2014, 9(1): e85902. DOI: 10.1371/journal.pone.0085902.
SHUKLA D, SAXENA S, PURUSHOTHAMAN J, et al . Hypoxic preconditioning with cobalt ameliorates hypobaric hypoxia induced pulmonary edema in rat[J ] . Eur J Pharmacol, 2011, 656(1/2/3): 101-109. DOI: 10.1016/j.ejphar.2011.01.038.
NEHRA S, BHARDWAJ V, BANSAL A, et al . Nanocurcumin accords protection against acute hypobaric hypoxia induced lung injury in rats[J ] . J Physiol Biochem, 2016, 72(4): 763-779. DOI: 10.1007/s13105-016-0515-3.
0
浏览量
142
下载量
0
CSCD
- 网络PDF下载
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
