基于蛋白质组学技术对竹叶青蛇毒染毒家兔模型血清蛋白质变化的检测及其意义

doi:10.13481/j.1671-587X.20220206

Abstract

Abstract: Objective

To investigate the expression of serum proteins of the snake venom model of Trimeresurus stejnegeri， and to reveal the expressions of marker proteins in the process of poisoning.

Methods

Twelve Japanese big ear rabbits， male or female， were randomly divided into sham operation group and model group， with 6 rabbits in each group. The rabbits in model group were given intramuscular injection of 20 mg·kg^－1Trimeresurus stejnegeri snake venom， and the rabbits in sham operation group were given the same volume of normal saline injection； four hours later， the rabbits in two groups were anesthetized with sodium pentobarbital，the blood samples were obtained from the hearts of rabbits， and the blood serum was isolated. The differential proteins in serum of the rabbits in two groups were analyzed by tandem mass tag（TMT） combined with high performance liquid chromatography-tandem mass spectrometry （HPLC-TMS） quantitative proteomics technology. The biological changes of the differential proteins were analyzed by Proteome Discover 2.4 and other databases.

Results

The sham operation group and the experimental group were divided into two distinct clusters of differential proteins on the coordinate diagram of PCA analysis. A total of 199 differential proteins were identified and analyzed， including 139 up-regulated proteins； the top 5 up-regulated proteins were troponin I type 2，fast skeletal muscle（TNNI2）， parvalbum alpha（PV）， enolase2（ENO2），myoglobin（MB） and creatine kinase（CK）；there were 60 down-regulated proteins；the top 5 down-regulated proteins were apolipoprotein C1（APOC1），cinggulin（CGN）， proprotein invertase Bacillus subtilis proteinase 9（PCSK9） and transfer cobalamin protein 1（TCN1）.The differential proteins involved in the extracellular regulatory function were the most， followed by calcium ion regulation protein and degradation regulation proteins.

Conclusion

The expression levels of antigen processing and presentation function-related proteins change significantly during the process of infection of Trimeresurus stejnegeri venom， and CK and solute carrier fannily 16 member1（SLC16A1） could be used as candidate targets of post-infection injury.

Key words: Trimeresurus stejnegeri venom, TJMT labeled quantitative proteomics, Serum differentially expressed proteins, Creatine kinase, Solute carrier family 16 member 1

CLC Number:

R994

Weilian YANG,Shijun WANG,Fanghua SHEN,Yong ZHANG,Fuwei CHEN,Qiuxiang SU,Chao SHI,Qinyao YU,Tao CHEN. Detection of serum protein changes in rabbit model poisoned by Trimeresurus stejnegeri snake venom based on proteomics and its significance[J].Journal of Jilin University(Medicine Edition), 2022, 48(2): 308-316.

Figures/Tables 7

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Tab 1

Tab 2

Fig. 2

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Fig. 5

References 24

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Accession	Description of protein	Protein symbol	P value	FC
P02643	Troponin Ⅰ type 2, fast skeletal muscle	TNNI2	0.011 1	33.28
P02624	Parvalbumin alpha	PVALB	0.003 7	20.58
A0A5F9C342	2-phospho-D-glycerate hydro-lyase	ENO2	0.001 0	18.16
P02170	Myoglobin	MB	0.006 6	14.58
P00563	Creatine kinase	CK	0.000 2	13.99
P08507	Glycerol-3-phosphate dehydrogenase cytoplasmic	GPD1	0.010 8	12.88
A0A5F9CRZ7	Keratin 15	KRT15	0.023 2	12.27
Q28641	Myosin-4	MYH4	0.013 1	9.98
G1SZ91	Fatty acid binding protein 5	FABP5	0.011 6	9.60
A0A5F9C2V9	Syndecan 4	SDC4	0.004 3	9.29
A0A5F9D578	LIM domain binding 3	LDB3	0.005 8	8.96
P02586	Troponin C, skeletal muscle	TNNC2	0.006 8	8.75
G1U7S4	Phosphoglycerate mutase	PGAM2	0.000 2	8.06
P25704	Beta-enolase	ENO3	0.000 2	6.80
P00489	Glycogen phosphorylase, muscle form	PYGM	0.000 2	6.74
A0A5F9C4E2	Aldo-keto reductase family 1 member B1	AKR1B1	0.044 4	6.28
A0A5F9C3R6	Asparaginase-like protein 1	ASRGL1	<0.01	5.94
P11974	Pyruvate kinase PKM	PKM	<0.01	4.98
P00883	Fructose-bisphosphate aldolase A	ALDOA	0.007 7	4.74
G8ZF20	Alpha-globin 1	HBA1	0.039 6	4.71
G1U685	Chromogranin B	CHGB	0.002 2	4.35
P25230	Antimicrobial protein CAP18	CAP18	0.000 7	4.25
A0A5F9CU78	Nidogen 2	NID2	0.000 4	4.21
G1SNC4	Adenylate kinase isoenzyme 1	AK1	0.012 9	4.21
P00939	Triosephosphate isomerase	TPI1	0.001 8	4.01
A0A5F9DVL7	Tetraspanin	TSPAN8	0.000 2	3.51
G1T821	Fibrinogen like 1	FGL1	0.000 3	3.47
G1TCC1	Solute Carrier Family 16 Member 1	SLC16A1	0.020 2	3.43
A0A5F9D6D6	Collagen alpha-2(I) chain	COL1A2	0.014 8	3.42
G1TBW9	Dynein light chain	DYNLL2	0.023 8	3.41
FC:Foldchange.

Accession	Description of Protein	Protein symbol	P-value	FC
P33047	Apolipoprotein C-1	APOC1	0.029 2	0.122 5
B7NZD9	Cingulin (Predicted)	CGN	0.034 8	0.260 5
B6S6L6	Apolipoprotein C-Ⅰ	apoCⅠ	0.023 5	0.294 2
G1U461	Proprotein convertase 9	PCSK9	0.000 8	0.294 7
G1TA71	Transcobalamin 1	TCN1	0.000 9	0.376 8
P55057	Apolipoprotein C-4	APOC4	0.028 8	0.403 8
A0A5F9D253	Serpin family A member 7	SERPINA7	0.023 2	0.437 0
G1SJM1	Apolipoprotein H	CEP112	0.020 1	0.444 9
G1U9S1	Complement factor properdin	CFP	0.000 1	0.454 8
G1T5K3	Apolipoprotein A-Ⅱ	APOA2	0.013 8	0.462 7
P14461	Fibrinogen alpha chain	FGA	0.004 2	0.466 6
G1SQC7	Pleckstrin	PLEK	0.027 0	0.487 3
G1TFU9	Galectin-3-binding protein	LGALS3BP	0.000 1	0.498 1
G1SE06	Platelet-activating factor acetylhydrolase	PLA2G7	0.005 5	0.522 0
G1U754	Histidine-rich glycoprotein	HRG	0.000 1	0.587 3
G1U9R4	Apolipoprotein B	APOB	0.012 0	0.597 1
A0A5F9CI66	Ceruloplasmin	CP	0.007 9	0.618 5
G1SH58	Carboxypeptidase N subunit 1	CPN1	0.010 7	0.620 3
G1TRF0	Serglycin	SRGN	0.023 2	0.623 3
G1TCW1	Transferrin receptor protein 1	TFRC	0.026 1	0.645 6
G1SGV9	Proteasome subunit beta	PSMB8	0.019 4	0.649 8
P15196	Sex hormone-binding globulin	SHBG	0.017 6	0.649 9
G1TET0	Carboxypeptidase B2	CPB2	0.001 7	0.661 1
G1SQU1	Proteasome subunit beta	PSMB10	0.027 1	0.683 0
G1SER2	Glycoprotein Ib platelet subunit alpha	GP1BA	0.046 2	0.697 1
B7NZD7	Proteasome subunit beta	PSMB4	0.032 3	0.709 8
A0A5F9C295	Proteasome subunit alpha type	PSMA6	0.032 4	0.710 6
G1SZ14	Proteasome subunit alpha type-3	PSMA3	0.028 7	0.719 9
G1U415	Vitronectin	VTN	0.007 5	0.721 9
B7NZD9	Cingulin (Predicted)	CGN	0.034 8	0.726 0
FC:Foldchange.

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Detection of serum protein changes in rabbit model poisoned by Trimeresurus stejnegeri snake venom based on proteomics and its significance

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