The role of palmitoylated NOD2 in animal model of hemorrhagic shock

doi:10.11958/20212779

Abstract

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

CLC Number:

R441.9

Figures/Tables 7

References 18

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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^＊＊

The role of palmitoylated NOD₂ in animal model of hemorrhagic shock

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