棕榈酰化修饰的NOD2在失血性休克动物模型中的作用

doi:10.11958/20212779

天津医药 ›› 2022, Vol. 50 ›› Issue (10): 1050-1055.doi: 10.11958/20212779

棕榈酰化修饰的NOD₂在失血性休克动物模型中的作用

谷子¹(), 唐勇¹, 周程继¹, 郑强²^,^△()

1 成都市第二人民医院急诊科（邮编610000）
2 成都市第一人民医院急诊科

收稿日期:2022-01-04 修回日期:2022-06-06 出版日期:2022-10-15 发布日期:2022-10-20
通讯作者: 郑强 E-mail:3142848@qq.com;402857358@qq.com
作者简介:谷子（1975）,男,副主任医师,主要从事急诊医学方面研究。E-mail： 3142848@qq.com
基金资助:
四川省卫生健康委员会普及应用项目(20PJ190)

The role of palmitoylated NOD₂ in animal model of hemorrhagic shock

GU Zi¹(), TANG Yong¹, ZHOU Chengji¹, ZHENG Qiang²^,^△()

1 Department of Emergency, Chengdu Second People's Hospital, Chengdu 610000, China
2 Department of Emergency, Chengdu First People's Hospital

Received:2022-01-04 Revised:2022-06-06 Published:2022-10-15 Online:2022-10-20
Contact: ZHENG Qiang E-mail:3142848@qq.com;402857358@qq.com

谷子, 唐勇, 周程继, 郑强. 棕榈酰化修饰的NOD₂在失血性休克动物模型中的作用[J]. 天津医药, 2022, 50(10): 1050-1055.

GU Zi, TANG Yong, ZHOU Chengji, ZHENG Qiang. The role of palmitoylated NOD₂ in animal model of hemorrhagic shock[J]. Tianjin Medical Journal, 2022, 50(10): 1050-1055.

摘要/Abstract

摘要：

目的探讨核苷酸结合寡聚化结构域2（NOD₂）在失血性休克及缺血再灌注时对肺损伤的作用及机制。方法 10只NOD₂基因敲除SD大鼠,经失血性休克模型制备后设为对照组（复苏初期予以生理盐水）。30只普通SD大鼠经失血性休克动物模型制备后,按随机数字表法分为3组：空白组（复苏初期予以生理盐水）、棕榈酰化修饰酶（PAT）抗体组（anti-PAT组,复苏初期予以PAT抗体）及NOD₂抗体组（anti-NOD₂组,复苏初期予以NOD₂抗体）,每组10只。经复苏24 h采集血液标本后将其处死并采集肺组织。在实验开始、休克初期及再灌注后2 h分别进行血气分析。检测肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β、IL-6及肺组织丙二醛（MDA）、髓过氧化物酶（MPO）水平。免疫印迹法检测肺组织胞核高迁移率族蛋白B（HMGB1）、棕榈酰化转移蛋白（ZDHHC5）及NOD₂表达水平。荧光显微镜及光镜下观察肺组织病理切片。结果 240 min时空白组氧分压[p（O₂）]明显低于其他3组（P<0.05）;30 min及240 min时空白组乳酸水平高于其他3组（P<0.05）。空白组TNF-α、IL-1β、IL-6、MDA及MPO水平最高（P<0.05）,且对照组低于anti-PAT组和anti-NOD₂组。空白组HMGB1、NOD₂和ZDHHC5表达量最高（P<0.05）,且对照组HMGB1和NOD₂表达量低于anti-PAT组和anti-NOD₂组（P<0.05）。空白组棕榈酰化修饰的NOD₂的免疫荧光表达量明显高于对照组、anti-PAT组及anti-NOD₂组。空白组较对照组、anti-PAT组及anti-NOD₂组肺泡内皮及肺泡组织破坏明显,炎性细胞浸润明显。结论在失血性休克及缺血再灌注动物模型中,NOD₂在ZDHHC5的调控下产生棕榈酰化修饰的NOD₂,从而促使TNF-α、IL-1β、IL-6、MDA及MPO的释放,损伤肺组织。

关键词: Nod2信号接头蛋白质, 脂化作用, 休克,出血性, 疾病模型,动物

Abstract:

Objective To investigate the effect and mechanism of nucleotide binding oligomerization domain 2 (NOD₂) on lung injury during hemorrhagic shock and ischemia-reperfusion. Methods Ten NOD₂ knockout SD rats were set as the control group after the preparation of hemorrhagic shock model (normal saline was given at the initial stage of resuscitation). Thirty ordinary SD rats were randomly divided into three groups: the blank group (normal saline was given at the initial stage of resuscitation), the palmitoyl modification enzyme (PAT) antibody group (PAT antibody was given at the beginning of resuscitation) and the NOD₂ antibody group (NOD₂ antibody was given at the beginning of resuscitation), with 10 rats in each group. After 24 hours of resuscitation, blood samples were collected, and lung tissues were collected. Blood gas analysis was performed at the beginning of the experiment, at the beginning of shock and 2 hours after reperfusion. Plasma tumor necrosis factor (TNF)-α, interleukin (IL) -1β, IL-6, malondialdehyde (MDA) and myeloperoxidase (MPO) in lung tissue were detected. High mobility group B (HMGB1), palmitoylated transfer protein (ZDHHC5) and NOD₂ were detected by Western blot assay. The pathological sections of lung tissue were observed under fluorescence microscope and light microscope. Results The oxygen partial pressure [p(O₂)] at 240 min was significantly lower in the blank group than that in the other three groups (P<0.05). The lactate levels at 30 min and 240 min were significantly higher in the blank group than those in the other three groups (P<0.05). TNF- α, IL-1β, IL-6, MDA and MPO levels were the highest in the blank group (P<0.05),and which was lower in the control group than that of the anti-PAT group and the anti-NOD₂ group. The expression levels of HMGB1, NOD₂ and ZDHHC5 were the highest in the blank group (P<0.05), and the expression levels of HMGB1 and NOD₂ were lower in the control group than those in the anti-PAT group and anti-NOD₂ group (P<0.05). The immunofluorescence expression of palmitoylated NOD₂ was significantly higher in the blank group than that in the control group, anti-PTA group and anti-NOD₂ group. Compared with the control group, the alveolar endothelium and alveolar tissue were significantly damaged and the inflammatory cell infiltration was obvious in the blank group, anti-PTA group and anti-NOD₂ group. Conclusion In hemorrhagic shock and ischemia-reperfusion animal models, NOD₂ produces palmitoylated NOD₂ under the regulation of ZDHHC5, thereby promoting the release of TNF-α, IL-1β, IL-6, MDA and MPO, resulting in lung inju

Key words: Nod2 signaling adaptor protein, lipoylation, shock, hemorrhagic, disease models, animal

中图分类号:

R441.9

图/表 7

参考文献 18

[1]	LI M, LI G, YU B, et al. Activation of hypoxia-inducible factor-1α via succinate dehydrogenase pathway during acute lung injury induced by trauma/hemorrhagic shock[J]. Shock, 2020, 53(2):208-216. doi: 10.1097/SHK.0000000000001347.
[2]	SHAYLOR R, GAVISH L, YANIV G, et al. Early maladaptive cardiovascular responses are associated with mortality in a porcine model of hemorrhagic shock[J]. Shock, 2020, 53(4):485-492. doi: 10.1097/SHK.0000000000001401.
[3]	RAO T W, SHEN Y H, ZHAO X G, et al. Effect of oxygen supplement during targeted temperature management on acute lung injury in the early stage of traumatic hemorrhagic shock[J]. Eur J Inflamm, 2020, 18:1-7. doi: 10.1177/2058739220930448.
[4]	NAPIER R J, LEE E J, VANCE E E, et al. Nod2 deficiency augments Th17 responses and exacerbates autoimmune arthritis[J]. J Immunol, 2018, 201(7):1889-1898. doi: 10.4049/jimmunol.1700507.
[5]	CHEN W, ZHENG D, MOU T, et al. Tle1 attenuates hepatic ischemia/reperfusion injury by suppressing NOD2/NF-κB signaling[J]. Biosci Biotechnol Biochem, 2020, 84(6):1176-1182. doi: 10.1080/09168451.2020.1735928.
[6]	WILLIAMS H, CAMPBELL L, CROMPTON R A, et al. Microbial host interactions and impaired wound healing in mice and humans:defining a role for BD14 and NOD2[J]. J Investig Dermatol, 2018, 138(10):2264-2274. doi: 10.1016/j.jid.2018.04.014.
[7]	CARUSO R, NÚÑEZ G. Innate immunity:ER stress recruits NOD1 and NOD2 for delivery of inflammation[J]. Curr Biol, 2016, 26(12):R508-R511. doi: 10.1016/j.cub.2016.05.021.
[8]	SIDDIQUE I, MUSTAFA A S, KHAN I, et al. Detection of mutations in NOD2/CARD15 gene in Arab patients with Crohn's disease[J]. Saudi J Gastroenterol, 2021, 27(4):240-248. doi: 10.4103/SJG.SJG_582_20.
[9]	SUZUKI K, SHINKAI H, YOSHIOKA G, et al. NOD2 genotypes affect the symptoms and mortality in the porcine circovirus 2-spreading pig population[J]. Genes(Basel), 2021, 12(9):1424. doi: 10.3390/genes12091424.
[10]	FANI MALEKI A, CISBANI G, LAFLAMME N, et al. Selective immunomodulatory and neuroprotective effects of a NOD2 receptor agonist on mouse models of multiple sclerosis[J]. Neurotherapeutics, 2021, 18(2):889-904. doi: 10.1007/S13311-020-00998-0.
[11]	MUKHERJEE T, HOVINGH E S, FOERSTER E G, et al. NOD1 and NOD2 in inflammation,immunity and disease[J]. Arch Biochem Biophys, 2019, 670:69-81. doi: 10.1016/j.abb.2018.12.022.
[12]	CARLOS D, PÉREZ M M, LEITE J A, et al. NOD2 deficiency promotes intestinal CD4⁺ T lymphocyte imbalance,metainflammation,and aggravates type 2 diabetes in murine model[J]. Front Immunol, 2020, 11:1265. doi: 10.3389/fimmu.2020.01265.
[13]	KIM Y K, SHIN J S, NAHM M H. NOD-like receptors in infection,immunity,and diseases[J]. Yonsei Med, 2016, 57(1):5-14. doi: 10.3349/ymj.2016.57.1.5.
[14]	Jakopin Ž, Corsini E. THP-1 cells and pro-inflammatory cytokine production:An In vitro tool for functional characterization of NOD1/NOD2 antagonists[J]. Int J Mol Sci, 2019, 20(17):4265. doi: 10.3390/ijms20174265.
[15]	LU Y, ZHENG Y, COYAUD E, et al. Palmitoylation of NOD1 and NOD2 is required for bacterial sensing[J]. Sci, 2019, 366(6464):460-467. doi: 10.1126/science.aau6391.
[16]	CHAN K L, TAM T H, BOROUMAND P, et al. Circulating NOD1 activators and hematopoietic NOD1 contribute to metabolic inflammation and insulin resistance[J]. Cell Rep, 2017, 18(10):2415-2426. doi: 10.1016/j.celrep.2017.02.027.
[17]	ZHAO C, ZHAO W. NLRP3 inflammasome-a key player in antiviral responses[J]. Front Immunol, 2020, 11:211. doi: 10.3389/fimmu.2020.00211.
[18]	LIU L, ZHANG L, ZHAO S, et al. Non-canonical Notch signaling regulates actin remodeling in cell migration by activating PI3K/AKT/Cdc42 pathway[J]. Front Pharmacol, 2019, 10:370. doi: 10.3389/fphar.2019.00370.

组别	pH						p（O₂）（mmHg）						p（CO₂）（mmHg）
组别	0 min		30 min		240 min		0 min		30 min		240 min		0 min		30 min		240 min
空白组	7.39±0.01		7.21±0.01		7.39±0.01		97.1±1.1		80.2±1.1		82.0±2.5		40.7±1.2		30.5±1.1		40.9±2.1
对照组	7.40±0.01		7.21±0.01		7.40±0.01		97.0±0.8		81.4±1.8		92.8±1.4^a		40.0±0.9		31.1±1.4		42.3±1.7
anti-PAT组	7.39±0.01		7.23±0.01^ab		7.39±0.01		97.9±1.1		81.2±1.5		94.6±1.5^ab		40.3±0.9		30.7±1.4		42.4±1.9
anti-NOD₂组	7.40±0.01		7.22±0.02		7.39±0.01		97.4±1.1		80.8±1.6		94.0±1.9^a		40.4±0.9		31.0±0.9		42.9±2.0^a
F	0.655		2.692^＊		1.380		1.555		1.165		101.480^＊＊		1.160		0.396		1.849^＊
组别		HCO₃^-（mmol/L ）				BE（mmol/L）						Lac（mmol/L）
组别		0 min	30 min	240 min		0 min		30 min		240 min		0 min		30 min		240 min
空白组		23.2±0.4	13.0±0.8	20.8±0.4		3.5±0.3		-16.4±0.6		-7.2±0.6		1.5±0.2		15.2±1.1		14.9±1.4
对照组		23.2±0.7	13.3±1.0	21.4±0.8		3.6±0.3		-16.0±0.7		-7.2±0.5		1.5±0.2		12.4±0.7^a		5.3±0.6^a
anti-PAT组		23.4±0.5	13.1±0.7	21.9±1.2^a		3.8±0.3^a		-16.2±0.7		-6.9±0.6		1.5±0.2		12.6±0.8^a		5.0±0.6^a
anti-NOD₂组		23.4±0.5	13.2±1.1	21.2±1.2		3.7±0.3		-16.2±0.7		-7.1±0.4		1.5±0.2		12.8±0.6^a		5.3±0.4^a
F		0.414	0.193	2.509^＊		1.605^＊		0.701		0.697		0.025		27.054^＊＊		326.881^＊＊

组别	pH						p（O₂）（mmHg）						p（CO₂）（mmHg）
组别	0 min		30 min		240 min		0 min		30 min		240 min		0 min		30 min		240 min
空白组	7.39±0.01		7.21±0.01		7.39±0.01		97.1±1.1		80.2±1.1		82.0±2.5		40.7±1.2		30.5±1.1		40.9±2.1
对照组	7.40±0.01		7.21±0.01		7.40±0.01		97.0±0.8		81.4±1.8		92.8±1.4^a		40.0±0.9		31.1±1.4		42.3±1.7
anti-PAT组	7.39±0.01		7.23±0.01^ab		7.39±0.01		97.9±1.1		81.2±1.5		94.6±1.5^ab		40.3±0.9		30.7±1.4		42.4±1.9
anti-NOD₂组	7.40±0.01		7.22±0.02		7.39±0.01		97.4±1.1		80.8±1.6		94.0±1.9^a		40.4±0.9		31.0±0.9		42.9±2.0^a
F	0.655		2.692^＊		1.380		1.555		1.165		101.480^＊＊		1.160		0.396		1.849^＊
组别		HCO₃^-（mmol/L ）				BE（mmol/L）						Lac（mmol/L）
组别		0 min	30 min	240 min		0 min		30 min		240 min		0 min		30 min		240 min
空白组		23.2±0.4	13.0±0.8	20.8±0.4		3.5±0.3		-16.4±0.6		-7.2±0.6		1.5±0.2		15.2±1.1		14.9±1.4
对照组		23.2±0.7	13.3±1.0	21.4±0.8		3.6±0.3		-16.0±0.7		-7.2±0.5		1.5±0.2		12.4±0.7^a		5.3±0.6^a
anti-PAT组		23.4±0.5	13.1±0.7	21.9±1.2^a		3.8±0.3^a		-16.2±0.7		-6.9±0.6		1.5±0.2		12.6±0.8^a		5.0±0.6^a
anti-NOD₂组		23.4±0.5	13.2±1.1	21.2±1.2		3.7±0.3		-16.2±0.7		-7.1±0.4		1.5±0.2		12.8±0.6^a		5.3±0.4^a
F		0.414	0.193	2.509^＊		1.605^＊		0.701		0.697		0.025		27.054^＊＊		326.881^＊＊

组别	TNF-α（ng/L）	IL-1β（ng/L）	IL-6（ng/L）	MDA（μmol/g）	MPO（μg/L）
空白组	216.4±12.0	152.6±6.1	103.1±6.0	167.3±10.7	226.1±12.5
对照组	34.7±2.4^a	22.7±1.4^a	22.2±3.3^a	31.0±2.1^a	50.1±3.6^a
anti-PAT组	141.6±3.8^ab	86.5±5.7^ab	61.4±5.5^ab	88.2±6.0^ab	127.0±6.4^ab
anti-NOD₂组	153.8±4.7^abc	91.0±4.9^abc	63.0±5.2^ab	103.6±8.0^abc	138.2±7.0^abc
F	1 224.163^＊＊	1 168.905^＊＊	424.840^＊＊	573.928^＊＊	797.415^＊＊

组别	TNF-α（ng/L）	IL-1β（ng/L）	IL-6（ng/L）	MDA（μmol/g）	MPO（μg/L）
空白组	216.4±12.0	152.6±6.1	103.1±6.0	167.3±10.7	226.1±12.5
对照组	34.7±2.4^a	22.7±1.4^a	22.2±3.3^a	31.0±2.1^a	50.1±3.6^a
anti-PAT组	141.6±3.8^ab	86.5±5.7^ab	61.4±5.5^ab	88.2±6.0^ab	127.0±6.4^ab
anti-NOD₂组	153.8±4.7^abc	91.0±4.9^abc	63.0±5.2^ab	103.6±8.0^abc	138.2±7.0^abc
F	1 224.163^＊＊	1 168.905^＊＊	424.840^＊＊	573.928^＊＊	797.415^＊＊

组别	HMGB1	NOD₂	ZDHHC5
空白组	2.10±0.04	1.26±0.12	1.39±0.11
对照组	0.75±0.07^a	0.19±0.03^a	1.17±0.16^a
anti-PAT组	1.26±0.20^ab	0.54±0.06^ab	0.56±0.10^ab
anti-NOD₂组	1.45±0.10^abc	0.65±0.06^abc	1.03±0.08^abc
F	221.503^＊＊	365.514^＊＊	90.836^＊＊

棕榈酰化修饰的NOD₂在失血性休克动物模型中的作用

The role of palmitoylated NOD₂ in animal model of hemorrhagic shock

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