1.Effectsofnucleusaccumbensablativesurgeryonamplitudeoflow-frequencyfluctuationsofbrain inheroindependentpatients:aresting-statefunctionalmagneticresonanceimagingstudy
Fan WANG ; Xiaohuai LI ; Feng HU ; Jing CHEN ; Wei LIU ; Jiajie CHEN ; Qiang LI ; Longxiao WEI ; Wei WANG
Journal of Practical Radiology 2019;35(7):1034-1037,1053
Objective Toinvestigatethechangesinbrainfunctionofheroindependentindividualsafternucleusaccumbens(NAC) ablativesurgery.Methods Twenty-fourheroindependentindividualsafterNACablativesurgery(NACgroup),27heroindependent individualsinshort-termabstinence(HDgroup)and32healthycontrol(HCgroup)wereincludedaresting-statefMRIstudy.Amplitudeoflow-frequencyfluctuations(ALFF)valuesofeachparticipantswerecalculated.DiffernecesinALFFvalueamongthethreegroupswere analyzed by One-Way ANOVA .Results Brain regions with statistically significant A L F F differences a m ong the three groups included left cerebellum,rightposteriorcentralgyrus,leftcaudatenucleus,rightmiddlefrontalgyrus,leftcuneusandbilateralinferiortemporal gyrusI.ntherightcentralposteriorgyrus,theALFFvalueofHDgroupwaslowerthanthatofNACandHCgroupI.ntheleftcerebellum,the ALFFvalueofHDgroupwashigherthanthatofNACandHCgrouprespectivelyI.nbilateralinferiortemporalgyrusandleftcaudatenucleus, ALFFvaluesofNACandHDgroupswerelowerthanHCgroup.ALFFvaluesofNACandHDgroupswerehigherthanHCgroup inrightmedialfrontalgyrusandleftcuneus.Conclusion InheroindependentindividualsafterNACablativesurgery,theALFFvaluesofleft cerebellumandrightposteriorcentralgyrustendtobenormal.NodifferenceinALFFvalueinregionsofdefaultmodenetworkbetweenNACand HDgroups.
2.Establishment and Evaluation of Rat Model of Acute Myocardial Infarction in Coronary Heart Disease with Qi and Yin Deficiency Syndrome Based on Sleep Deprivation Combined with Coronary Artery Ligation
Yali SHI ; Yunxiao GAO ; Qiuyan ZHANG ; Yue YUAN ; Xiaoxiao CHEN ; Longxiao HU ; Junguo REN ; Jianxun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):30-40
ObjectiveTo explore the construction and evaluation methods of a rat model of acute myocardial infarction(AMI) with Qi and Yin deficiency syndrome established by sleep deprivation combined with coronary artery ligation. MethodsThirty-six SD rats were randomly divided into a normal group(n=6), a myocardial infarction group(model A group, n=10), an acute sleep deprivation+myocardial infarction group(model B group, n=10), and a chronic sleep deprivation+myocardial infarction group(model C group, n=10) according to body weight. Rats in the normal group were not treated, rats in the model A group underwent only ligation of the left anterior descending coronary artery, rats in the model B group were sleep deprived for 96 h and then underwent ligation of the left anterior descending coronary artery, and rats in the model C group were sleep deprived for an additional 48 h each week with a 24 h rest period as one cycle for three weeks on the basis of the model B group. After coronary artery ligation in the model C group, the first week was defined as the starting point of the first sleep deprivation cycle, and indexes were tested weekly for rats in each group for 3 weeks. Electrocardiogram was used to determine the ligation of the left anterior descending coronary artery in rats, and small animal echocardiography was used to evaluate the cardiac function. The levels of serum creatine kinase(CK), creatine kinase isoenzyme(CK-MB), lactate dehydrogenase(LDH), cardiac troponin T(cTnT), interleukin-18(IL-18), and tumor necrosis factor-α(TNF-α) were detected by biochemical assays, and hematoxylin-eosin(HE) staining was used to evaluate the pathological changes of myocardial tissue in rats. The syndrome indicators of Qi and Yin deficiency were evaluated by general state and body weight, grip strength, facial temperature, paw temperature, rectal temperature, salivary flow rate, open field test, tongue color[red(R), green(G), and blue(B)] values, pulse amplitude changes, and enzyme-linked immunosorbent assay(ELISA) for the detection of expression levels of cyclic adenosine monophosphate(cAMP), cyclic guanosine monophosphate(cGMP), rat serum corticotropin-releasing factor(CRF), adrenocorticotropic hormone(ACTH), triiodothyronine(T3), tetraiodothyronine(T4), and corticosterone(CORT) in serum. ResultsIn terms of disease indicators, compared with the normal group, the ST segment of the electrocardiogram in each model group was significantly elevated, the echocardiographic parameters were decreased, the contents of myocardial enzymes and inflammatory factors were increased(P<0.01), and the myocardial tissue in the infarcted area was significantly damaged. In terms of syndrome indicators, compared with the normal group, the body weight of rats in the model B and C groups decreased at each time point, the grip strength of each model group decreased, the total distance traveled and the number of entries into the center in the open field test decreased, the immobility time increased, the facial and rectal temperatures of rats in the model B and C groups increased, the salivary flow rate of each model group decreased, the tongue color was bright red or light, the tongue body was dry or smooth like a mirror, lacking of moisture sensation, the R, G and B values of the tongue surface increased, the pulse amplitude changes decreased, and the contents of T3 and T4 increased, while the expressions of cAMP, CRF, ACTH and CORT in the model B and C groups increased(P<0.05, P<0.01). ConclusionContinuous sleep deprivation for 96 h in a multi-platform method combined with coronary artery ligation can construct a rat model of AMI with Qi and Yin deficiency syndrome, and the syndrome manifestations can be maintained for 3 weeks.
3.Establishment and Evaluation of Mouse Model of Ischemic Heart Disease with Qi and Yin Deficiency Syndrome Based on Proteomics
Qiuyan ZHANG ; Ying LI ; Yunxiao GAO ; Longxiao HU ; Yue YUAN ; Xiaoxiao CHEN ; Yali SHI ; Junguo REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):52-61
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.
4.Establishment and Evaluation of Rat Model of Acute Myocardial Infarction in Coronary Heart Disease with Qi and Yin Deficiency Syndrome Based on Sleep Deprivation Combined with Coronary Artery Ligation
Yali SHI ; Yunxiao GAO ; Qiuyan ZHANG ; Yue YUAN ; Xiaoxiao CHEN ; Longxiao HU ; Junguo REN ; Jianxun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):30-40
ObjectiveTo explore the construction and evaluation methods of a rat model of acute myocardial infarction(AMI) with Qi and Yin deficiency syndrome established by sleep deprivation combined with coronary artery ligation. MethodsThirty-six SD rats were randomly divided into a normal group(n=6), a myocardial infarction group(model A group, n=10), an acute sleep deprivation+myocardial infarction group(model B group, n=10), and a chronic sleep deprivation+myocardial infarction group(model C group, n=10) according to body weight. Rats in the normal group were not treated, rats in the model A group underwent only ligation of the left anterior descending coronary artery, rats in the model B group were sleep deprived for 96 h and then underwent ligation of the left anterior descending coronary artery, and rats in the model C group were sleep deprived for an additional 48 h each week with a 24 h rest period as one cycle for three weeks on the basis of the model B group. After coronary artery ligation in the model C group, the first week was defined as the starting point of the first sleep deprivation cycle, and indexes were tested weekly for rats in each group for 3 weeks. Electrocardiogram was used to determine the ligation of the left anterior descending coronary artery in rats, and small animal echocardiography was used to evaluate the cardiac function. The levels of serum creatine kinase(CK), creatine kinase isoenzyme(CK-MB), lactate dehydrogenase(LDH), cardiac troponin T(cTnT), interleukin-18(IL-18), and tumor necrosis factor-α(TNF-α) were detected by biochemical assays, and hematoxylin-eosin(HE) staining was used to evaluate the pathological changes of myocardial tissue in rats. The syndrome indicators of Qi and Yin deficiency were evaluated by general state and body weight, grip strength, facial temperature, paw temperature, rectal temperature, salivary flow rate, open field test, tongue color[red(R), green(G), and blue(B)] values, pulse amplitude changes, and enzyme-linked immunosorbent assay(ELISA) for the detection of expression levels of cyclic adenosine monophosphate(cAMP), cyclic guanosine monophosphate(cGMP), rat serum corticotropin-releasing factor(CRF), adrenocorticotropic hormone(ACTH), triiodothyronine(T3), tetraiodothyronine(T4), and corticosterone(CORT) in serum. ResultsIn terms of disease indicators, compared with the normal group, the ST segment of the electrocardiogram in each model group was significantly elevated, the echocardiographic parameters were decreased, the contents of myocardial enzymes and inflammatory factors were increased(P<0.01), and the myocardial tissue in the infarcted area was significantly damaged. In terms of syndrome indicators, compared with the normal group, the body weight of rats in the model B and C groups decreased at each time point, the grip strength of each model group decreased, the total distance traveled and the number of entries into the center in the open field test decreased, the immobility time increased, the facial and rectal temperatures of rats in the model B and C groups increased, the salivary flow rate of each model group decreased, the tongue color was bright red or light, the tongue body was dry or smooth like a mirror, lacking of moisture sensation, the R, G and B values of the tongue surface increased, the pulse amplitude changes decreased, and the contents of T3 and T4 increased, while the expressions of cAMP, CRF, ACTH and CORT in the model B and C groups increased(P<0.05, P<0.01). ConclusionContinuous sleep deprivation for 96 h in a multi-platform method combined with coronary artery ligation can construct a rat model of AMI with Qi and Yin deficiency syndrome, and the syndrome manifestations can be maintained for 3 weeks.
5.Establishment and Evaluation of Mouse Model of Ischemic Heart Disease with Qi and Yin Deficiency Syndrome Based on Proteomics
Qiuyan ZHANG ; Ying LI ; Yunxiao GAO ; Longxiao HU ; Yue YUAN ; Xiaoxiao CHEN ; Yali SHI ; Junguo REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):52-61
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.
6.Establishment and Evaluation of Mouse Model of Cerebral Infarction with Qi and Yin Deficiency Syndrome Based on Metabolomics
Yue YUAN ; Yunxiao GAO ; Qiuyan ZHANG ; Xiaoxiao CHEN ; Yali SHI ; Longxiao HU ; Jianxun LIU ; Junguo REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):62-71
ObjectiveTo investigate the preparation method of a mouse model of cerebral infarction with Qi and Yin deficiency syndrome induced by streptozotocin(STZ) combined with the photochemical method, and to evaluate the biological basis of the established model. MethodsForty C57B6/J mice were randomly divided into the normal and model groups, with 20 mice in each group. The normal group received no treatment, while the model group was injected intraperitoneally with 55 mg·kg-1 of STZ once a day for 5 days. Fourteen days post-STZ induction, 10 mice from the normal group were randomly taken into the photochemical group, while 10 mice from the model group were randomly taken into the STZ+photochemical group. Rose Bengal solution injection combined with 520 nm laser irradiation was used to cause thrombosis and induce cerebral infarction in mice. Syndrome indexes for Qi and Yin deficiency were assessed by general state observation, body weight, grip strength, rectal temperature, behavioral experiments, energy metabolism, tongue color[red(R), green(G), blue(B)] values, adenosine triphosphate(ATP) content, corticotropin-releasing factor(CRF) and triiodothyronine(T3) levels. The pathological changes of cerebral infarction in mice were evaluated by detecting serum superoxide dismutase(SOD), interleukin-1β(IL-1β), IL-6, and tumor necrosis factor-α(TNF-α) levels in combination with Bederson score. Finally, the endogenous metabolites in mice were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS), and multivariate statistical analysis was performed by partial least squares-discriminant analysis(PLS-DA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). The data filtering criteria were set as variable importance in the projection(VIP) value> 1, fold change(FC)<0.8 or FC>1.2, P<0.05, to obtain differential metabolites. Then MetaboAnalyst 3.0 was utilized for pathway enrichment analysis of the differential metabolites, aiming to explore the metabolic profile changes and biological basis of mice with Qi and Yin deficiency syndrome of cerebral infarction. ResultsRegarding the syndrome indicators, compared with the normal group, the mice in the model group had lower body weight, higher rectal temperature, lower limb motor ability and energy metabolism efficiency, lower ATP content, lower R, G and B values of the tongue surface, and lower speed of blood glucose regression(P<0.05, P<0.01). As for the disease indicators, compared with the normal group, the Bederson scores of the photochemical group and the STZ+photochemical group increased, the grip strength decreased, the SOD level decreased, and the levels of inflammatory factors increased(P<0.05). The results of metabolomics showed that a good separation pattern of components was observed among mice in each group, with significant differences in components. Identification of MS data revealed a total of 44 differential metabolites in mice with Qi and Yin deficiency syndrome of cerebral infarction. Among them, 32 metabolites were up-regulated, mainly including triglycerides, diglycerides, phospholipids, and ceramides. And 12 metabolites were down-regulated, mainly including amino acid and phosphate metabolites. Pathway enrichment analysis of the above differential metabolites indicated that the metabolic pathways were mainly enriched in folate biosynthesis, terpenoid skeleton biosynthesis, glycerophospholipid metabolism, vitamin B6 metabolism, glycerolipid metabolism and sphingolipid metabolism. These pathways were involved in multiple processes such as lipid transport, insulin resistance, and energy metabolism. ConclusionThe method of STZ injection combined with photochemical induction can successfully establish a mouse model of cerebral infarction with Qi and Yin deficiency syndrome, and intervene in vivo processes such as folate biosynthesis, glycerophospholipid metabolism, and glycerolipid metabolism.
7.Establishment and Evaluation of Mouse Model of Cerebral Infarction with Qi and Yin Deficiency Syndrome Based on Metabolomics
Yue YUAN ; Yunxiao GAO ; Qiuyan ZHANG ; Xiaoxiao CHEN ; Yali SHI ; Longxiao HU ; Jianxun LIU ; Junguo REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):62-71
ObjectiveTo investigate the preparation method of a mouse model of cerebral infarction with Qi and Yin deficiency syndrome induced by streptozotocin(STZ) combined with the photochemical method, and to evaluate the biological basis of the established model. MethodsForty C57B6/J mice were randomly divided into the normal and model groups, with 20 mice in each group. The normal group received no treatment, while the model group was injected intraperitoneally with 55 mg·kg-1 of STZ once a day for 5 days. Fourteen days post-STZ induction, 10 mice from the normal group were randomly taken into the photochemical group, while 10 mice from the model group were randomly taken into the STZ+photochemical group. Rose Bengal solution injection combined with 520 nm laser irradiation was used to cause thrombosis and induce cerebral infarction in mice. Syndrome indexes for Qi and Yin deficiency were assessed by general state observation, body weight, grip strength, rectal temperature, behavioral experiments, energy metabolism, tongue color[red(R), green(G), blue(B)] values, adenosine triphosphate(ATP) content, corticotropin-releasing factor(CRF) and triiodothyronine(T3) levels. The pathological changes of cerebral infarction in mice were evaluated by detecting serum superoxide dismutase(SOD), interleukin-1β(IL-1β), IL-6, and tumor necrosis factor-α(TNF-α) levels in combination with Bederson score. Finally, the endogenous metabolites in mice were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS), and multivariate statistical analysis was performed by partial least squares-discriminant analysis(PLS-DA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). The data filtering criteria were set as variable importance in the projection(VIP) value> 1, fold change(FC)<0.8 or FC>1.2, P<0.05, to obtain differential metabolites. Then MetaboAnalyst 3.0 was utilized for pathway enrichment analysis of the differential metabolites, aiming to explore the metabolic profile changes and biological basis of mice with Qi and Yin deficiency syndrome of cerebral infarction. ResultsRegarding the syndrome indicators, compared with the normal group, the mice in the model group had lower body weight, higher rectal temperature, lower limb motor ability and energy metabolism efficiency, lower ATP content, lower R, G and B values of the tongue surface, and lower speed of blood glucose regression(P<0.05, P<0.01). As for the disease indicators, compared with the normal group, the Bederson scores of the photochemical group and the STZ+photochemical group increased, the grip strength decreased, the SOD level decreased, and the levels of inflammatory factors increased(P<0.05). The results of metabolomics showed that a good separation pattern of components was observed among mice in each group, with significant differences in components. Identification of MS data revealed a total of 44 differential metabolites in mice with Qi and Yin deficiency syndrome of cerebral infarction. Among them, 32 metabolites were up-regulated, mainly including triglycerides, diglycerides, phospholipids, and ceramides. And 12 metabolites were down-regulated, mainly including amino acid and phosphate metabolites. Pathway enrichment analysis of the above differential metabolites indicated that the metabolic pathways were mainly enriched in folate biosynthesis, terpenoid skeleton biosynthesis, glycerophospholipid metabolism, vitamin B6 metabolism, glycerolipid metabolism and sphingolipid metabolism. These pathways were involved in multiple processes such as lipid transport, insulin resistance, and energy metabolism. ConclusionThe method of STZ injection combined with photochemical induction can successfully establish a mouse model of cerebral infarction with Qi and Yin deficiency syndrome, and intervene in vivo processes such as folate biosynthesis, glycerophospholipid metabolism, and glycerolipid metabolism.
8.Establishment and Evaluation of Animal Model of Acute Myocardial Infarction with Syndrome of Qi and Yin Deficiency
Yunxiao GAO ; Qiuyan ZHANG ; Juqin PENG ; Hao GUO ; Xiaoxiao CHEN ; Wei HAO ; Longxiao HU ; Yali SHI ; Junguo REN ; Jianxun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):134-142
ObjectiveTo explore the establishment and evaluation methods of the rat model of acute myocardial infarction (AMI) in coronary heart disease with the syndrome of Qi and Yin deficiency by sleep deprivation (SD) combined with isoproterenol (ISO) and preliminarily explore its biological basis. MethodForty SD rats were assigned into normal (no treatment), SD (treatment in modified multi-platform water environment for 96 h), ISO (subcutaneous injection of ISO at 100 mg·kg-1 once every other day for a total of 2 times), and SD+ISO (injection of 100 mg·kg-1 ISO after SD for 72 h and 96 h) groups. The cardiac function was detected by small animal echocardiography. The serum levels of creatine kinase (CK), creatine kinase isoenzyme (CK-MB), lactate dehydrogenase (LDH), and cardiac troponin T (cTnT) were measured by biochemical methods. The pathological changes of the myocardial tissue were observed by hematoxylin-eosin staining. The general state, body weight, grip strength, body temperature, behaviors in open field test, serum levels of cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), cAMP/cGMP ratio, red (R), green (G), blue (B) values of the tongue surface, and pulse amplitude were observed and measured to evaluate the modeling results. Enzyme-linked immunosorbent assay was employed to determine the serum levels of interleukin-18 (IL-18), tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD), malondialdehyde (MDA), corticotropin-releasing factor (CRF), adrenocorticotropic hormone (ACTH), triiodothyronine (T3), tetraiodothyronine (T4), cluster of differentiation 4 (CD4), and cluster of differentiation 8 (CD8). ResultIn terms of disease indicators, the ISO and SD+ISO groups had lower cardiac function indicators than the normal group (P<0.01). The levels of CK, CM-MB, LDH and cTnT elevated in each model group compared with the normal group (P<0.01). The pathological changes of myocardial tissue were obvious in the ISO and SD+ISO groups. In terms of syndrome indicators, compared with the normal group, the SD and SD+ISO groups showed decreased body weight at each time point (P<0.01), and the ISO group showed decreased body weight at the time points of 48 h and 72 h (P<0.05, P<0.01). The paw temperature and rectal temperature increased in the SD group (P<0.01). The model groups showed weakened grasp strength, lowered R, G, and B values of the tongue surface (P<0.01), prolonged immobility time (P<0.01), reduced total distance and number of entering the central area (P<0.01), decreased average speed (P<0.05, P<0.01), and increased cAMP and cGMP (P<0.05, P<0.01). The cAMP/cGMP ratio was increased in the SD+ISO group (P<0.01), and the pulse amplitude was decreased in the SD and SD+ISO groups (P<0.01). In terms of serological indicators,compared with the normal group, the levels of IL-18, TNF-α, SOD and MDA were significantly increased in the ISO and SD+ISO groups (P<0.01), the CRF, ACTH, CORT, T3, T4, CD4 and CD8 in the model groups were increased (P<0.05, P<0.01). ConclusionSleep deprivation for 96 h combined with high-dose ISO can successfully establish a rat model of acute myocardial infarction in coronary heart disease with the syndrome of Qi and Yin deficiency. The model evaluation system can be built with disease indicators of western medicine, histopathological indicators, macroscopic indicators of traditional Chinese medicine, and serological indicators.
9.Establishment and Evaluation of Rat Model of Myocardial Ischemia-reperfusion Injury with Phlegm and Blood Stasis Blocking Collaterals Syndrome Based on Metabolomics
Longxiao HU ; Jiabei GAO ; Weihao MA ; Jieming LU ; Yunxiao GAO ; Yue YUAN ; Qiuyan ZHANG ; Xiaoxiao CHEN ; Yali SHI ; Jianxun LIU ; Junguo REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):41-51
ObjectiveTo explore the feasibility, evaluation methods and metabolic differences of high-fat diet(HFD) combined with myocardial ischemia-reperfusion injury(MIRI) to establish a rat model of myocardial ischemia-reperfusion with phlegm and blood stasis blocking collaterals syndrome(PBSBCS). MethodsThirty-two SD rats were randomly divided into the sham operation, HFD, MIRI, and MIRI+HFD groups. Rats in the sham operation and MIRI groups were fed a standard diet(regular chow), while the HFD and MIRI+HFD groups received a HFD for 10 weeks. Rats in the MIRI and MIRI+HFD groups underwent myocardial ischemia-reperfusion surgery, while the sham operation group underwent only thread placement without ligation. Cardiac function was assessed via small-animal echocardiography, including left ventricular ejection fraction(EF), left ventricular fractional shortening(FS), cardiac output(CO), and stroke volume(SV). Serum levels of creatine kinase(CK), CK-MB, triglyceride(TG), total cholesterol(TC), high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C), lactate dehydrogenase(LDH), endothelin-1(ET-1), endothelial nitric oxide synthase(eNOS), tumor necrosis factor-α(TNF-α), interleukin-18(IL-18), oxidized LDL(ox-LDL), and cardiac troponin T(cTnT) were measured by biochemical assays and enzyme-linked immunosorbent assay(ELISA). Myocardial histopathology was evaluated via hematoxylin-eosin(HE) staining, while myocardial infarction and no-reflow area were assessed using 2,3,5-triphenyltetrazolium chloride(TTC), Evans blue, and thioflavin staining. Changes in syndrome characteristics[body weight, tongue surface red-green-blue [RGB] values, and pulse amplitude] of PBSBCS were recorded. Serum differential metabolites were analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS). ResultsCompared with the sham operation group, the HFD and MIRI+HFD groups showed significant increases in body weight(P<0.01), RGB values and pulse amplitude decreased in the HFD, MIRI and MIRI+HFD groups, TC, TG, LDL-C and ox-LDL levels increased in the HFD and MIRI+HFD groups, while HDL-C decreased. Blood perfusion peak time and myocardial no-reflow area increased, serum eNOS level decreased, and CK-MB, LDH, and cTnT activities increased in the HFD, MIRI and MIRI+HFD groups(P<0.05, P<0.01). Whole blood viscosity was increased in the HFD group at medium shear rate, and in the MIRI and MIRI+HFD groups at low, medium and high shear rates(P<0.05, P<0.01). Platelet aggregation rate increased in the MIRI and MIRI+HFD groups, accompanied by elevated ET-1, TNF-α, and IL-18 levels, reduced cardiac function indices, expanded myocardial no-reflow and infarction areas, and increased serum CK, CK-MB, LDH, and cTnT activities(P<0.05, P<0.01). Compared with the MIRI group, the HFD and MIRI+HFD groups showed significant increase in body weight, TC, TG, LDL-C and ox-LDL levels, and significant decrease in HDL-C content(P<0.01). The MIRI+HFD group showed decrease in RGB values and pulse amplitude, and an increase in whole blood viscosity, platelet aggregation, blood perfusion peak time, myocardial no-reflow and infarction areas, elevated ET-1, TNF-α and IL-18 levels, decreased eNOS content, EF and SV, increased serum CK, CK-MB and cTnT activities, and worsened myocardial pathology(P<0.05). Compared with the HFD group, the MIRI+HFD group showed similar aggravated trends(P<0.05, P<0.01). Metabolomics results showed that 34 potential biomarkers involving 13 common metabolic pathways were identified in the MIRI+HFD group compared with the sham operation group. ConclusionThe MIRI group resembles blood stasis syndrome in hemodynamics and myocardial injury, and the HFD group mirrors phlegm-turbidity syndrome in lipid profiles and tongue characteristics. While the MIRI+HFD group aligns with PBSBCS in comprehensive indices, effectively simulating clinical features of coronary heart disease(CHD), which can be used for the evaluation of the pathological mechanism and pharmacodynamics of CHD with PBSBCS.
10.Establishment and Evaluation of Rat Model of Myocardial Ischemia-reperfusion Injury with Phlegm and Blood Stasis Blocking Collaterals Syndrome Based on Metabolomics
Longxiao HU ; Jiabei GAO ; Weihao MA ; Jieming LU ; Yunxiao GAO ; Yue YUAN ; Qiuyan ZHANG ; Xiaoxiao CHEN ; Yali SHI ; Jianxun LIU ; Junguo REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):41-51
ObjectiveTo explore the feasibility, evaluation methods and metabolic differences of high-fat diet(HFD) combined with myocardial ischemia-reperfusion injury(MIRI) to establish a rat model of myocardial ischemia-reperfusion with phlegm and blood stasis blocking collaterals syndrome(PBSBCS). MethodsThirty-two SD rats were randomly divided into the sham operation, HFD, MIRI, and MIRI+HFD groups. Rats in the sham operation and MIRI groups were fed a standard diet(regular chow), while the HFD and MIRI+HFD groups received a HFD for 10 weeks. Rats in the MIRI and MIRI+HFD groups underwent myocardial ischemia-reperfusion surgery, while the sham operation group underwent only thread placement without ligation. Cardiac function was assessed via small-animal echocardiography, including left ventricular ejection fraction(EF), left ventricular fractional shortening(FS), cardiac output(CO), and stroke volume(SV). Serum levels of creatine kinase(CK), CK-MB, triglyceride(TG), total cholesterol(TC), high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C), lactate dehydrogenase(LDH), endothelin-1(ET-1), endothelial nitric oxide synthase(eNOS), tumor necrosis factor-α(TNF-α), interleukin-18(IL-18), oxidized LDL(ox-LDL), and cardiac troponin T(cTnT) were measured by biochemical assays and enzyme-linked immunosorbent assay(ELISA). Myocardial histopathology was evaluated via hematoxylin-eosin(HE) staining, while myocardial infarction and no-reflow area were assessed using 2,3,5-triphenyltetrazolium chloride(TTC), Evans blue, and thioflavin staining. Changes in syndrome characteristics[body weight, tongue surface red-green-blue [RGB] values, and pulse amplitude] of PBSBCS were recorded. Serum differential metabolites were analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS). ResultsCompared with the sham operation group, the HFD and MIRI+HFD groups showed significant increases in body weight(P<0.01), RGB values and pulse amplitude decreased in the HFD, MIRI and MIRI+HFD groups, TC, TG, LDL-C and ox-LDL levels increased in the HFD and MIRI+HFD groups, while HDL-C decreased. Blood perfusion peak time and myocardial no-reflow area increased, serum eNOS level decreased, and CK-MB, LDH, and cTnT activities increased in the HFD, MIRI and MIRI+HFD groups(P<0.05, P<0.01). Whole blood viscosity was increased in the HFD group at medium shear rate, and in the MIRI and MIRI+HFD groups at low, medium and high shear rates(P<0.05, P<0.01). Platelet aggregation rate increased in the MIRI and MIRI+HFD groups, accompanied by elevated ET-1, TNF-α, and IL-18 levels, reduced cardiac function indices, expanded myocardial no-reflow and infarction areas, and increased serum CK, CK-MB, LDH, and cTnT activities(P<0.05, P<0.01). Compared with the MIRI group, the HFD and MIRI+HFD groups showed significant increase in body weight, TC, TG, LDL-C and ox-LDL levels, and significant decrease in HDL-C content(P<0.01). The MIRI+HFD group showed decrease in RGB values and pulse amplitude, and an increase in whole blood viscosity, platelet aggregation, blood perfusion peak time, myocardial no-reflow and infarction areas, elevated ET-1, TNF-α and IL-18 levels, decreased eNOS content, EF and SV, increased serum CK, CK-MB and cTnT activities, and worsened myocardial pathology(P<0.05). Compared with the HFD group, the MIRI+HFD group showed similar aggravated trends(P<0.05, P<0.01). Metabolomics results showed that 34 potential biomarkers involving 13 common metabolic pathways were identified in the MIRI+HFD group compared with the sham operation group. ConclusionThe MIRI group resembles blood stasis syndrome in hemodynamics and myocardial injury, and the HFD group mirrors phlegm-turbidity syndrome in lipid profiles and tongue characteristics. While the MIRI+HFD group aligns with PBSBCS in comprehensive indices, effectively simulating clinical features of coronary heart disease(CHD), which can be used for the evaluation of the pathological mechanism and pharmacodynamics of CHD with PBSBCS.