1.Comparison of Wild and Cultivated Bupleurum scorzonerifolium Based on Traditional Quality Evaluation
Changsheng YUAN ; Feng ZHOU ; Xingyu LIU ; Yu SHI ; Yihan WANG ; Huaizhu LI ; Yongliang LI ; Shan GUAN ; Huaizhong GAO ; Yanmeng LIU ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):203-214
ObjectiveTo characterize the quality differences among different germplasm and introduced varieties of Bupleurum scorzonerifolium roots(BSR), and explore the underlying molecular mechanisms, providing a basis for high-quality production and quality control. MethodsWild BSR from Yulin(YLW) served as the quality reference, we conducted comparative analysis among YLW, locally domesticated wild germplasm in Yulin(YLC3), Daqing germplasm introduced and cultivated in Yulin(YLDQC3), and locally cultivated germplasm in Daqing(DQC3). A combination of traditional pharmacognostic methods and modern multi-omics analyses was employed, including macroscopic traits(appearance, odor), microscopic features(proportions of cork, phloem, xylem), cell wall component contents(hemicellulose, cellulose, lignin), carbohydrate contents(starch, water-soluble polysaccharides), marker compound contents(ethanol-soluble extracts, total saponins, liposoluble extracts, and saikosaponins A, B2, C, D), metabolomics, and transcriptomics, in order to systematically characterize quality differences and investigate molecular mechanisms among these samples. ResultsMacroscopically, Yulin-produced BSR(YLW, YLC3, YLDQC3) exhibited significantly greater weight, length, and upper and middle diameters than Daqing-produced BSR(DQC3). Odor-wise, YLW and YLC3 had a a fragrance taste, YLDQC3 had a rancid oil odor, and DQC3 had a sweet and fragrant taste. Microscopically, Yulin germplasm(YLW, YLC3) and Daqing germplasm(YLDQC3, DQC3) shared similar structural features, respectively. However, Yulin germplasm showed significantly higher proportions of cork and phloem, as well as stronger xylem vessel staining intensity compared to Daqing germplasm. Regarding various component contents, Yulin germplasm contained significantly higher levels of ethanol-soluble extracts, total saponins, and saikosaponins A, B2, C, D, while Daqing germplasm had significantly higher levels of hemicellulose, starch, and liposoluble extracts. After introduction to Yulin, the Daqing germplasm(YLDQC3) showed increased starch, water-soluble polysaccharides and liposoluble extracts contents, decreased cell wall component content, but no significant difference in other component contents. Metabolomics revealed that saponins and terpenes accumulated significantly in Yulin germplasm, while alcohols and aldehydes accumulated predominantly in Daqing germplasm. Transcriptomics indicated similar gene expression patterns within the same germplasm but specificity between different germplasms. Integrative metabolomic-transcriptomic analysis identified 145 potential key genes associated with the saikosaponin biosynthesis pathway, including one acetyl-coenzyme A(CoA) acetyltransferase gene(ACAT), one 3-hydroxy-3-methylglutaryl-coenzyme A synthase gene(HMGS), two hydroxymethylglutaryl-CoA(HMG-CoA) reductase genes(HMG), one phosphomevalonate kinase gene(PMK), one 1-deoxy-D-xylose-5-phosphate synthase gene(CLA), one hydroxymethylbuten-1-aldol synthase gene(HDR), two farnesyl pyrophosphate synthase genes(FPPS), one squalene synthase gene(SQS), one β-amyrin synthase gene(BAS), 102 cytochrome P450(CYP450) gene family members, and 32 uridine diphosphate-glucuronosyltransferase(UGT) gene family members. ConclusionAmong the three cultivated types, YLC3 most closely resembles YLW in appearance, microscopic features, contents of major bioactive constituents, metabolomic and transcriptomic profiles. Yulin germplasm exhibits superior saponin synthesis capability compared to Daqing germplasm, and Yulin region is more suitable for the growth of B. scorzonerifolium. Based on these findings, it is recommended that artificial cultivation in northern Shaanxi and similar regions utilize the local Yulin germplasm source cultivated for at least three years.
2.Pathogen spectrum of diarrheal disease surveillance in Fengxian District, Shanghai, 2013‒2023
Meihua LIU ; Yuan ZHUANG ; Xiaohong XIE ; Hongwei ZHAO ; Yuan SHI ; Lijuan DING ; Yi HU ; Lixin TAO
Shanghai Journal of Preventive Medicine 2025;37(4):336-341
ObjectiveTo investigate the pathogenic spectrum and epidemiological characteristics of diarrheal disease in Fengxian District of Shanghai, and to provide scientific basis for the prevention and control of diarrheal diseases. MethodsBasic information of the initial adult cases visited diarrheal disease surveillance sentinel hospital in Fengxian District, Shanghai, was collected from August 2013 to 2023, and fecal samples were collected at 1∶5 sampling intervals to isolate and identify 5 kinds of diarrheagenic Escherichia coli (DEC), Salmonella (SAL), Vibrio parahaemolyticus, Campylobacter, Vibrio cholerae, Shigella and Yersinia enterocolitica (YE). Simultaneously, nucleic acid detection was performed for 3 kinds of rotavirus, 2 kinds of norovirus, intestinal adenovirus, astrovirus and sapovirus. ResultsA total of 1 861 cases of newly diagnosed diarrheal disease were reported, with the peak in July to August. Additionally, 704 surveillance samples were detected, with a total positive detection rate of 50.57%. The detection rates of bacterial, viral and mixed infection were 25.14%, 21.02% and 4.40%, respectively. Among the pathogens detected, DEC accounted for the highest (17.61%, 124/704), followed by norovirus (16.48%, 116/704), rotavirus (6.39%, 45/704), SAL (5.97%, 42/704) and Campylobacter (3.84%, 27/704). DEC detected were mainly enteroaggregative Escherichia coli and enterotoxigenic Escherichia coli, with no detection of Vibrio cholerae, Shigella and YE. The highest total pathogen detection rate was observed from June to September, and the detection peaks of norovirus were from March to June and from October to December, whereas that of DEC was from June to October. The detection rate of rotavirus peaked from January to February, but which was not detected between 2020‒2023. The SAL positive rate peak was in September, whereas that of Campylobacter was from July to September. ConclusionThe main pathogens detected in Fengxian District from 2013‒2019 are DEC, norovirus, rotavirus, SAL and Campylobacter. Different pathogens have different detection peaks, with bacteria predominating in summer and viruses in winter and spring. Prevention and control measures should be carried out according to the epidemiological characteristics of different seasons.
3.Chemical consitituents and hypoglycemic activity of Qinhuai No. 1 Rehmannia glutinosa
Meng YANG ; Zhi-you HAO ; Xiao-lan WANG ; Chao-yuan XIAO ; Jun-yang ZHANG ; Shi-qi ZHOU ; Xiao-ke ZHENG ; Wei-sheng FENG
Acta Pharmaceutica Sinica 2025;60(1):205-210
Eight compounds were isolated and purified from the ethyl acetate part of 70% acetone extract of
4.Influence of parental smoking and household secondhand smoke exposure on adolescents smoking initiation
SU Qi, XU Luting, SHI Jianhui, ZENG Xinying, LIU Ying, QI Mingxin, CAO Yuan, LIU Shiwei
Chinese Journal of School Health 2025;46(3):339-342
Objective:
To explore the impact of household tobacco smoke exposure on adolescents attempted smoking behavior, so as to provide a reference for tobacco control policy formulation and evaluation.
Methods:
From September to November 2023, a stratified cluster random sampling method was employed to select 7 841 middle and high school students from 10 monitoring sites (districts/counties) in Beijing for a questionnaire survey. Rao-Scott Chi square test was used to assess differences in proportions across subgroups, and complex sampling design based multivariate Logistic regression analysis was conducted to explore the influence of parental smoking and secondhand smoke (SHS) exposure at home on adolescents attempted smoking behavior.
Results:
About 47.17% of adolescents reported to have at least one parent smoked, with 42.36% reported of having only the father smoked, 0.73% reported of having only the mother smoked, and 4.08% reported of having both parents smoked. About 34.66% of middle and high school students were reported SHS exposure at home in the past 7 days, with 10.98%, 4.79% and 18.89% reported SHS exposure for 1-2, 3-4 and 5-7 days. Compared to adolescents with non smoking parents, those with a smoking father or both smoking parents had higher rates of attempted smoking [ OR (95% CI )=1.45(1.06-1.98), 3.73(2.18-6.37), P < 0.05 ]. Compared to adolescents without SHS exposure at home in the past 7 days, those exposed for 3-4 or 5- 7 days had higher rates of attempted smoking [ OR (95% CI )=2.21(1.27- 3.84 ), 2.46(1.58-3.83), P <0.01].
Conclusions
Household tobacco smoke exposure is associated with adolescent attempted smoking behavior. Parents should quit smoking and prohibit smoking at home to create a smoke free environment for adolescents.
5.Analysis of Differences in Secondary Metabolites Between Dendrobium nobile Bionic Wild Cultivated on Epiphytic Stones and Trees Based on Widely Targeted Metabolomics
Yifan SHI ; Changqing ZHOU ; Jiaojiao WANG ; Lin CHEN ; Hongping CHEN ; Fu WANG ; Yuan HU ; Youping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):218-224
ObjectiveTo explore the differences in the accumulation of secondary metabolites of Dendrobium nobile cultivated on epiphytic stones and trees, so as to elucidate the scientific connotation of "only those that grow on stones has superior quality", and provide a direction for the cultivation and quality evaluation of D. nobile. MethodsUltra-performance liquid chromatography-triple quadrupole/linear ion trap mass spectrometry(UPLC-QTRAP-MS/MS)-based widely targeted metabolomics was used to detect the metabolites of D. nobile cultivated on epiphytic stones and trees. And the combination of principal component analysis(PCA), hierarchical cluster analysis(HCA), and orthogonal partial least squares-discriminant analysis(OPLS-DA) was performed for multivariate statistical analysis of metabolites. Differential metabolites were screened by variable importance in the projection(VIP) value≥1 and log2fold change(FC)≥1 or ≤-1, and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis was conducted. ResultsA total of 1 267 metabolites were identified in the stems of D. nobile from the two cultivation modes, dominated by flavonoids(292), phenolic acids(284), and alkaloids(189). Through OPLS-DA screening, 473 differential metabolites were obtained. Compared to epiphytic tree-cultivated D. nobile, epiphytic stone-cultivated D. nobile exhibited upregulation of flavonoids, phenolic acids, alkaloids, lignans and coumarins, while quinones and terpenoids were down-regulated. The differential metabolites mainly included flavonoid glycosides and alkaloids, and these differential metabolites significantly contributed to characterizing the two cultivation patterns. KEGG enrichment analysis revealed significant enrichment in pathways of flavone and flavonol biosynthesis, flavonoid biosynthesis, tyrosine metabolism, and phenylalanine metabolism in epiphytic stone-cultivated D. nobile. ConclusionEpiphytic stone cultivation is beneficial for the accumulation of phenolic acids, flavonoids, and alkaloids in D. nobile, indicating that the "only those that grow on stones has superior quality" documented in the materia medica has certain scientific basis, and the findings also provide a reference for quality evaluation and discrimination research between epiphytic stone and tree cultivated D. nobile.
6.Process Optimization and Health Risk Assessment of Calcined Haematitum Based on QbD Concept
Yue YANG ; Jingwei ZHOU ; Jialiang ZOU ; Guorong MEI ; Yifan SHI ; Lei ZHONG ; Jiaojiao WANG ; Xuelian GAN ; Dewen ZENG ; Xin CHEN ; Lin CHEN ; Hongping CHEN ; Shilin CHEN ; Yuan HU ; Youping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):187-196
ObjectiveTo investigate the processing technology of calcined Haematitum based on the concept of quality by design(QbD) and to assess its health risk. MethodsTaking whole iron content, Fe2+ dissolution content and looseness as critical quality attributes(CQAs), and calcination temperature, calcination time, spreading thickness and particle size as critical process parameters(CPPs) determined by the failure mode and effect analysis(FMEA), the processing technology of calcined Haematitum was optimized by orthogonal test combined with analytic hierarchy process-criteria importance through intercriteria correlation(AHP-CRITIC) hybrid weighting method. The contents of heavy metals and harmful elements were determined by inductively coupled plasma mass spectrometry, and the health risk assessment was carried out by daily exposure(EXP), target hazard quotient(THQ) and lifetime cancer risk(LCR), and the theoretical value of the maximum limit was deduced. ResultsThe optimal processing technology for calcined Haematitum was calcination at 650 ℃, calcination time of 1 h, particle size of 0.2-0.5 cm, spreading thickness of 1 cm, and vinegar quenching for 1 time[Haematitum-vinegar(10:3)]. The contents of 5 heavy metals and harmful elements in 13 batches of calcined Haematitum were all decreased with reductions of up to 5-fold. The cumulative THQ of 2 batches of samples was>1, while the cumulative THQ of all batches of Haematitum was>1. The LCR of As in 1 batches of Haematitum was 1×10-6-1×10-4, and the LCR of the rest was<1×10-6, and the LCRs of calcined Haematitum were all<1×10-6, indicating that the carcinogenic risk of calcined Haematitum was low, but special attention should still be paid to Haematitum medicinal materials. Preliminary theoretical values of the maximum limits of Cu, As, Cd, Pb and Hg were formulated as 1 014, 25, 17, 27, 7 mg·kg-1. ConclusionThe optimized processing technology of calcined Haematitum is stable and feasible, and the contents of heavy metals and harmful elements are reduced after processing. Preliminary theoretical values of the maximum limits of Cu, As, Cd, Pb and Hg are formulated to provide a scientific basis for the formulation of standards for the limits of harmful elements in Haematitum.
7.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.
8.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.
9.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.
10.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.


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