Establishment and Evaluation of Mouse Model of Ischemic Heart Disease with Qi and Yin Deficiency Syndrome Based on Proteomics
10.13422/j.cnki.syfjx.20251463
- VernacularTitle:基于蛋白质组学的缺血性心脏病气阴两虚证小鼠模型的建立与评价
- Author:
Qiuyan ZHANG
1
;
Ying LI
2
;
Yunxiao GAO
3
;
Longxiao HU
1
;
Yue YUAN
2
;
Xiaoxiao CHEN
2
;
Yali SHI
2
;
Junguo REN
1
Author Information
1. Beijing University of Chinese Medicine，Beijing 100029，China
2. Beijing Key Laboratory of Chinese Materia Pharmacology，Institute of Basic Medical Sciences， Xiyuan Hospital，China Academy of Chinese Medical Sciences，Beijing 100091，China
3. Natural History Museum of China，Beijing 100050，China
- Publication Type:Journal Article
- Keywords:
orthogonal test;
isoproterenol;
Qi and Yin deficiency syndrome;
ischemic heart disease;
mouse model;
proteomics
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2025;31(12):52-61
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the optimal construction method and the biological basis for establishing a mouse model of ischemic heart disease（IHD） with Qi and Yin deficiency syndrome by intraperitoneal injection of isoproterenol（ISO）. MethodsA total of 144 male C57BL/6J mice were randomly assigned into three normal groups and nine model groups according to body mass， with 12 mice in each group. The model groups 1， 4， and 7 were administered ISO via intraperitoneal injection at a dose of 5 mg·kg-1·d-1 for four consecutive days， the model groups 2， 5， and 8 received ISO at a dose of 10 mg·kg-1·d-1 for seven consecutive days， while the model groups 3， 6， and 9 were given ISO at a dose of 15 mg·kg-1·d-1 for 14 consecutive days. The normal groups were administered an equivalent volume of normal saline via intraperitoneal injection. After the modeling process， body mass， 24-hour food and water intake， grip strength， and spontaneous activity of the mice were measured. Cardiac function was assessed using echocardiography， the serum levels of norepinephrine（NE）， cyclic adenosine monophosphate（cAMP）， and cyclic guanosine monophosphate（cGMP） were determined via enzyme-linked immunosorbent assay（ELISA）. The content of adenosine triphosphate（ATP） in myocardial tissue was measured by biochemical analysis， while histopathological changes in myocardial tissue were observed via hematoxylin-eosin（HE） staining. An orthogonal experimental design was applied for intuitive analysis and variance analysis to screen the optimal modeling conditions of the mouse model of IHD with Qi and Yin deficiency syndrome. A data-dependent acquisition（DDA） proteomic technique was employed to quantitatively detect differentially expressed proteins in myocardial tissue between the optimal model group and the normal group. And bioinformatics analysis was conducted to explore the potential biological mechanisms underlying the Qi and Yin deficiency model of IHD. ResultsOrthogonal results showed that the injection cycle had a great influence on model establishment， and the optimal modeling condition was identified as intraperitoneal injection of ISO at 15 mg·kg-1·d-1 for 14 consecutive days. Under this condition， compared with the normal group， the model group demonstrated significant reductions in body mass， food intake， water intake， grip strength， total distance and average speed of exercise， ejection fraction（EF）， fractional shortening（FS）， serum levels of NE and cGMP， and myocardial ATP content（P<0.01）， while immobility time， cAMP level， and the cAMP/cGMP value were significantly increased（P<0.05， P<0.01）. HE staining results revealed that myocardial tissue in the model group had disordered cell arrangement， inflammatory cell infiltration， myocardial fiber rupture， and fibrous tissue proliferation. Proteomic analysis identified 141 differentially expressed proteins in the model group compared with the normal group， with 52 up-regulated and 89 down-regulated. Gene Ontology（GO） functional annotation and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis indicated that the cellular components（CC） were mainly related to mitochondria and the inner mitochondrial membrane， the biological processes（BP） were associated with complement activation， platelet activation， and responses to metal ions， suggesting that the potential functional pathways involved the complement and coagulation cascade， as well as porphyrin metabolism. ConclusionContinuous intraperitoneal injection of ISO at a dose of 15 mg·kg-1 for 14 days successfully establishes a mouse model of IHD with Qi and Yin deficiency syndrome， and the underlying mechanisms may be related to the regulation of iron ions by complement C3， C5 and Cp， and plays a role in the regulation through the BP of complement activation， platelet activation， and responses to metal ions， and the signaling pathways of the complement and coagulation cascade and porphyrin metabolism.
