抑制周细胞铁死亡对脓毒症大鼠肺血管屏障功能的保护作用

刘奕言; 张紫森; 包代琴; 王洪晨; 李涛; 刘良明

doi:10.16016/j.2097-0927.202205044

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抑制周细胞铁死亡对脓毒症大鼠肺血管屏障功能的保护作用

专题报道：战创伤休克救治 | 更新时间：2026-01-09

- 抑制周细胞铁死亡对脓毒症大鼠肺血管屏障功能的保护作用
- Protective effects of inhibiting pericyte ferroptosis on pulmonary vascular barrier function in septic rats
- 陆军军医大学学报 2022年44卷第20期页码：2037-2047
- 作者机构：
  
  1.400042 重庆，陆军特色医学中心: 野战外科研究部战伤休克与输血研究室，创伤、烧伤与复合伤国家重点实验室
  2.400042 重庆，陆军特色医学中心: 麻醉科
- 作者简介：
  
  [ 刘良明，E-mail: lmliu62@tmmu.edu.cn LIU Liangming, E-mail: lmliu62@tmmu.edu.cn
- 基金信息：
  
  国家自然科学基金重点项目(81830065)
- DOI：10.16016/j.2097-0927.202205044
  中图分类号： R322.35;R329.25;R631.2
- 收稿：2022-05-10，
  
  修回：2022-07-18，
  
  纸质出版：2022-10-30
- 稿件说明：
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刘奕言, 张紫森, 包代琴, 等. 抑制周细胞铁死亡对脓毒症大鼠肺血管屏障功能的保护作用[J]. 陆军军医大学学报, 2022,44(20):2037-2047.

Yiyan LIU, Zisen ZHANG, Daiqin BAO, et al. Protective effects of inhibiting pericyte ferroptosis on pulmonary vascular barrier function in septic rats[J]. Journal of Army Medical University, 2022, 44(20): 2037-2047.
刘奕言, 张紫森, 包代琴, 等. 抑制周细胞铁死亡对脓毒症大鼠肺血管屏障功能的保护作用[J]. 陆军军医大学学报, 2022,44(20):2037-2047. DOI： 10.16016/j.2097-0927.202205044.

Yiyan LIU, Zisen ZHANG, Daiqin BAO, et al. Protective effects of inhibiting pericyte ferroptosis on pulmonary vascular barrier function in septic rats[J]. Journal of Army Medical University, 2022, 44(20): 2037-2047. DOI： 10.16016/j.2097-0927.202205044.

摘要

目的

探讨周细胞铁死亡在脓毒症大鼠肺血管屏障功能损伤中的作用及铁死亡抑制剂(ferrostatin-1，Fer-1)对脓毒症大鼠肺血管屏障功能的保护作用。

方法

采用盲肠结扎穿孔术(cecal ligation and puncture，CLP)构建大鼠脓毒症模型。①将156只SD大鼠按随机数表法分为假手术组(Sham组)、盲肠结扎穿孔术组(CLP组)和铁死亡抑制剂组(Fer-1组)；Fer-1组在CLP术前1 h经腹腔注射Fer-1(10 mg/kg)，流式细胞学实验取假手术组大鼠肺单细胞悬液增设空白

未染色组(Blank组)。取肺微小静脉观察不同处理组大鼠肺血管周细胞数量变化，观察各组大鼠肺血管通透性、肺湿干质量比以及肺病理结构变化；观察各组大鼠肺血管周细胞脂质过氧化物蓄积以及肺微小静脉脂质过氧化物(lipid peroxidation，LPO)含量和铁死亡标志性蛋白表达变化。②提取原代周细胞分为正常对照组(Normal组)、脂多糖刺激组(lipopolysaccharide

LPS组)和Fer-1组，Fer-1组用2 μmol/L Fer-1预处理周细胞12 h后，再用10 μg/mL LPS处理周细胞12 h。观察各组周细胞内ROS、Lipid ROS和Fe

含量以及各组细胞铁死亡标志性蛋白GPX4、COX2表达。

结果

动物实验: 肺血管周细胞内脂质过氧化物蓄积，周细胞大量丢失，肺血管发生严重渗漏，伊文思蓝大量外渗，肺结构严重破坏伴随大量炎性细胞浸润，肺湿干质量比显著上升，Fer-1可减轻肺血管周细胞铁死亡，减少肺血管周细胞丢失，减轻肺血管渗漏，改善肺病理结构，显著降低肺微小静脉周细胞内脂质过氧化物含量，减轻铁死亡。细胞实验: LPS显著降低周细胞活性，诱导周细胞内ROS、Lipid ROS和Fe

大量蓄积，周细胞GPX4表达明显降低，COX2表达明显升高；Fer-1预处理可显著恢复周细胞活性，降低细胞内ROS、Lipid ROS和Fe

水平，恢复周细胞GPX4表达以及降低COX2表达。

结论

脓毒症可引起肺血管周细胞铁死亡和丢失，导致肺血管平屏障功能受损，铁死亡抑制剂Fer-1可拮抗脓毒症后周细胞铁死亡，对脓毒症后肺血管屏障功能有重要保护作用。

Abstract

Objective

To investigate the role of pericyte ferroptosis in the injury of pulmonary vascular barrier in septic rats and identify the protective effect of ferrostatin-1 (Fer-1)

a ferroptosis inhibitor

on pulmonary vascular barrier function.

Methods

① Animal experiment: The septic model of rats was established by cecal ligation and puncture (CLP)

and 156 SD rats were randomly divided into sham group

CLP group and Fer-1 group (10 mg/kg Fer-1 intraperitoneally in 1 h before CLP). Single cell suspension of rat lung from the sham-operation group was obtained by flow cytometry as an unstained blank control. The abundance of pericytes in pulmonary venules

changes in pulmonary vascular permeability

lung wet to dry weight ratio and lung histopathology of rats in each group were observed. Meanwhile

the contents of lipid peroxidation (LPO) in pulmonary pericytes and venules

as well as the expression of ferroptosis marker proteins including glutathione peroxidase 4 (GPX4) and cyclooxygenase-2 (COX2) were also measured in each group. ②Cell experiment: Primary pericytes were isolated and divided into norma

l control group

lipopolysaccharide (LPS) group (treated with 10 μg/mL LPS for 12 h)

and Fer-1 group (pretreated with 2 μmol/L Fer-1 for 12 h before LPS stimulation). The contents of ROS

Lipid ROS and Fe

and the expression levels of GPX4 and COX2 in the pericytes of each group were investigated.

Results

In vivo

massive detachment of pericytes from pulmonary veins

serious leakage of pulmonary vessels

large quantities of Evans blue extravasation

severe damage of lung pathological structure were observed in the CLP group

accompanied with infiltration of massive inflammatory cells

increase in lung wet-dry weight ratio

and accumulation of LPO in pulmonary vascular pericytes

which indicated severe ferroptosis. However

Fer-1 pretreatment reduced the ferroptosis of pulmonary pericytes

decreased the loss of pericytes

alleviated pulmonary vascular leakage

improved lung structure

and greatly lowered the contents of LPO in the pericytes (

0.01).

In vitro

LPS significantly impaired pericytes viability and induced accumulations of ROS

Lipid ROS and Fe

in the pericytes. Meanwhile

LPS down-regulated the expression of GPX4 while up-regulated that of COX2. Administration of Fer-1 restored the pericytes viability and the expression of GPX4

and decreased the levels of intracellular ROS

Lipid ROS

and COX2 expression.

Conclusion

Sepsis may induce the ferroptosis and detachment of pulmonary pericytes

and then impair pulmonary vascular barrier function. The ferroptosis inhibitor Fer-1 can antagonize pulmonary pericytes ferroptosis and play a vital role in protecting pulmonary vascular barrier function.

关键词

Keywords

references

PRESCOTT H C, ANGUS D C. Enhancing recovery from sepsis: a review[J]. JAMA, 2018, 319(1): 62-75. DOI: 10.1001/jama.2017.17687.

HUANG M, CAI S L, SU J Q. The pathogenesis of sepsis and potential therapeutic targets[J]. Int J Mol Sci, 2019, 20(21): 5376. DOI: 10.3390/ijms20215376.

GOTTS J E, MATTHAY M A. sepsis: pathophysiology and clinical management[J]. Brit Med J, 2016, 353: i1585. DOI: 10.1136/bmj.i1585.

ABRAHAM E. Efficacy and safety of monoclonal antibody to human tumor necrosis factor alpha in patients with sepsis syndrome. A randomized, controlled, double-blind, multicenter clinical trial. TNF-alpha MAb sepsis Study Group[J]. JAMA, 1995, 273(12): 934-941. DOI: 10.1001/jama.273.12.934.

ANNANE D, RENAULT A, BRUN-BUISSON C, et al . Hydrocortisone plus fludrocortisone for adults with septic shock[J ] . N Engl J Med, 2018, 378(9): 809-818. DOI: 10.1056/nejmoa1705716.

LEVY M M, EVANS L E, RHODES A. The surviving Sepsis campaign bundle: 2018 update[J ] . Intensive Care Med, 2018, 44(6): 925-928. DOI: 10.1007/s00134-018-5085-0.

ZHANG Y Y, NING B T. Signaling pathways and intervention therapies in Sepsis [J ] . Signal Transduct Target Ther, 2021, 6(1): 407. DOI: 10.1038/s41392-021-00816-9.

ARULKUMARAN N, POLLEN S, GRECO E, et al . Renal tubular cell mitochondrial dysfunction occurs despite preserved renal oxygen delivery in experimental septic acute kidney injury[J ] . Crit Care Med, 2018, 46(4): e318-e325. DOI: 10.1097/CCM.0000000000002937.

PAN P, WANG X T, LIU D W. The potential mechanism of mitochondrial dysfunction in septic cardiomyopathy[J]. J Int Med Res, 2018, 46(6): 2157-2169. DOI: 10.1177/0300060518765896.

YU H C, YANG C C, JIAN L, et al . Sulfasalazine-induced ferroptosis in breast cancer cells is reduced by the inhibitory effect of estrogen receptor on the transferrin receptor[J ] . Oncol Rep, 2019, 42(2): 826-838. DOI: 10.3892/or.2019.7189.

CHEN L Y, LI X X, LIU L B, et al . Erastin sensitizes glioblastoma cells to temozolomide by restraining xCT and cystathionine-γ-lyase function[J ] . Oncol Rep, 2015, 33(3): 1465-1474.DOI: 10.3892/or.2015.3712.

XIAO Z, KONG B, FANG J, et al. Ferrostatin-1 alleviates lipopolysaccharide-induced cardiac dysfunction[J]. Bioengineered, 2021, 12(2): 9367-9376. DOI: 10.1080/21655979.2021.2001913.

张紫森, 刘良明. 周细胞对脓毒症大鼠血管反应性的保护作用[J]. 第三军医大学学报, 2019, 41(21): 2029-2034. DOI: 10.16016/j.1000-5404.201907039.

ZHANG Z S, LIU L M. Role and mechanism of pericyte in protection of vascular reactivity in septic rats[J]. J Third Mil Med Univ, 2019, 41(21): 2029-2034. DOI: 10.16016/j.1000-5404.201907039.

王洪晨, 周赫男, 郑丹阳, 等. 周细胞微囊泡对脓毒症大鼠肺损伤的保护作用[J]. 第三军医大学学报, 2021, 43(13): 1185-1191. DOI: 10.16016/j.1000-5404.202101031.

WANG H C, ZHOU H N, ZHENG D Y, et al . Pericyte-derived microvesicles protects septic rats against lung injury[J ] . J Third Mil Med Univ, 2021, 43(13): 1185-1191. DOI: 10.16016/j.1000-5404.202101031.

GÖRITZ C, DIAS D O, TOMILIN N, et al . A pericyte origin of spinal cord scar tissue[J ] . Science, 2011, 333(6039): 238-242. DOI: 10.1126/science.1203165.

ATTWELL D, MISHRA A, HALL C N, et al . What is a pericyte?[J ] . J Cereb Blood Flow Metab, 2016, 36(2): 451-455. DOI: 10.1177/0271678X15610340.

HUANG H. Pericyte-endothelial interactions in the retinal microvasculature[J]. Int J Mol Sci, 2020, 21(19): 7413. DOI: 10.3390/ijms21197413.

MÄE M A, HE L Q, NORDLING S, et al . Single-cell analysis of blood-brain barrier response to pericyte loss[J ] . Circ Res, 2021, 128(4): e46-e62. DOI: 10.1161/CIRCRESAHA.120.317473.

NEES S, WEISS D R, SENFTL A, et al . Isolation, bulk cultivation, and characterization of coronary microvascular pericytes: the second most frequent myocardial cell type in vitro [J ] . Am J Physiol Heart Circ Physiol, 2012, 302(1): H69-H84. DOI: 10.1152/ajpheart.00359.2011..

ZHOU H N, ZHENG D Y, WANG H C, et al . The protective effects of pericyte-derived microvesicles on vascular endothelial functions via CTGF delivery in sepsis[J ] . Cell Commun Signal, 2021, 19(1): 115. DOI: 10.1186/s12964-021-00795-y.

张亮, 廖勇群, 夏秦川, 等. 铁死亡调控信号通路以及在相关疾病中的研究进展[J]. 中国临床药理学与治疗学, 2022, 27(2): 227-234. DOI: 10.12092/j.issn.1009-2501.2022.02.015.

ZHANG L, LIAO Y Q, XIA Q C, et al . Ferroptosis regulatory signaling pathway and its research progress in related diseases[J ] . Chin J Clin Pharmacol Ther, 2022, 27(2): 227-234. DOI: 10.12092/j.issn.1009-2501.2022.02.015.

耿慧君, 张佩, 王国艳, 等. 铁死亡及其研究进展[J]. 生命的化学, 2021, 41(11): 2376-2382. DOI: 10.13488/j.smhx.20210259.

GENG H J, ZHANG P, WANG G Y, et al . Research progress of ferroptosis[J ] . Chem Life, 2021, 41(11): 2376-2382. DOI: 10.13488/j.smhx.20210259.

DIXON S J, LEMBERG K M, LAMPRECHT M R, et al . Ferroptosis: an iron-dependent form of nonapoptotic cell death[J ] . Cell, 2012, 149(5): 1060-1072. DOI: 10.1016/j.cell.2012.03.042.

YANG W S, STOCKWELL B R. Synthetic lethal screening identifies compounds activating iron-dependent, nonapoptotic cell death in oncogenic-RAS-harboring cancer cells[J]. Chem Biol, 2008, 15(3): 234-245. DOI: 10.1016/j.chembiol.2008.02.010.

YAGODA N, VON RECHENBERG M, ZAGANJOR E, et al . RAS-RAF-MEK-dependent oxidative cell death involving voltage-dependent anion channels[J ] . Nature, 2007, 447(7146): 864-868. DOI: 10.1038/nature05859.

LIU Q J, WU J, ZHANG X F, et al . Iron homeostasis and disorders revisited in the sepsis[J ] . Free Radic Biol Med, 2021, 165: 1-13. DOI: 10.1016/j.freeradbiomed.2021.01.025.

LI N, WANG W, ZHOU H, et al . Ferritinophagy-mediated ferroptosis is involved in sepsis-induced cardiac injury[J ] . Free Radic Biol Med, 2020, 160: 303-318. DOI: 10.1016/j.freeradbiomed.2020.08.009.

WEI S S, BI J B, YANG L F, et al . Serum irisin levels are decreased in patients with sepsis, and exogenous irisin suppresses ferroptosis in the liver of septic mice[J ] . Clin Transl Med, 2020, 10(5): e173. DOI: 10.1002/ctm2.173.

HE R Y, LIU B H, XIONG R, et al . Itaconate inhibits ferroptosis of macrophage via Nrf2 pathways against sepsis-induced acute lung injury[J ] . Cell Death Discov, 2022, 8(1): 43. DOI: 10.1038/s41420-021-00807-3.

SIDDALL E, KHATRI M, RADHAKRISHNAN J. Capillary leak syndrome: etiologies, pathophysiology, and management[J]. Kidney Int, 2017, 92(1): 37-46. DOI: 10.1016/j.kint.2016.11.029.

MOK G, HENDIN A, REARDON P, et al . Macrocirculatory and microcirculatory endpoints in sepsis resuscitation[J ] . J Intensive Care Med, 2021, 36(12): 1385-1391. DOI: 10.1177/0885066620982585.

JOFFRE J, HELLMAN J, INCE C, et al . Endothelial responses in sepsis[J ] . Am J Respir Crit Care Med, 2020, 202(3): 361-370. DOI: 10.1164/rccm.201910-1911TR.

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