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.Advances in the JAK2/STAT3 signaling pathway and its inhibitors in diffuse large B cell lymphoma
Chuanyang LU ; Qiuni CHEN ; Yuye SHI ; Yuan DENG ; Tingting JI ; Zhengyuan LIU ; Chunling WANG ; Liang YU
China Pharmacy 2026;37(5):682-688
Abnormal activation of the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway is involved in the pathogenesis of diffuse large B-cell lymphoma (DLBCL). In recent years, inhibitors targeting JAK2 and STAT3 have emerged as promising therapeutic candidates in DLBCL. This review summarizes the efficacy and safety profiles of JAK2 inhibitors (e.g., ruxolitinib) and STAT3 inhibitors (direct small-molecule inhibitors, the antisense oligonucleotide, and proteolysis targeting chimeras, etc.) in preclinical models and clinical trials. Accumulating evidence indicates that JAK2 and STAT3 inhibitors exhibit antitumor activity and are generally well tolerated in a subset of DLBCL patients. Meanwhile, the development of novel drug delivery systems has significantly enhanced the stability, bioavailability, and targeting ability of the compounds. Furthermore, JAK2 and STAT3 inhibitors may exhibit synergistic effects when combined with other therapy strategies (such as combinations with B-cell receptor signaling pathway inhibitors, immunomodulators, or other targeted drugs). However, current clinical applications are still in their early stages. Future research should concentrate on precision treatment strategies based on the genetic subtyping of DLBCL, and further refine the delivery systems for inhibitors as well as combination drug regimens to improve clinical outcomes.
3.Mechanism of MEK/Ras/Raf/ERK Signaling Pathway Modulated by Mimenghua Prescription on Inflammatory Response in Dry Eye Animal Model
Shi TAN ; Pei LIU ; Yuan ZHONG ; Sainan TIAN ; Pengfei JIANG ; Genyan QIN ; Qinghua PENG ; Jun PENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):211-221
ObjectiveThis paper aims to investigate the effects and mechanism of Mimenghua prescription in modulating the mitogen-activated protein kinase kinase (MEK)/rat sarcoma viral oncogene homolog (Ras)/rapidly accelerated fibrosarcoma kinase (Raf)/extracellular signal-regulated kinase (ERK) signaling pathway to inhibit inflammatory responses in a dry eye animal model. MethodsA total of 60 C57BL/6J mice (eight weeks old, half male and half female) were used in the experiment. Ten mice were randomly selected as the blank control group, while the remaining 50 were exposed to a controlled dry system and received instillation of 0.2% benzalkonium chloride (BAC) into the eyes for four weeks to establish a dry eye mouse model. After successful modeling, the mice were randomly divided into five groups: Model group, sodium hyaluronate group, and Mimenghua prescription groups with low dose (4.83 g·kg-1), medium dose (9.67 g·kg-1), and high dose (19.34 g·kg-1). The mice in the model group received an equal volume of normal saline via gavage for four weeks. The mice in the sodium hyaluronate group received instillation of sodium hyaluronate eye drops twice daily for 14 consecutive days. The tear secretion volume, tear film break-up time (TBUT), and corneal fluorescein staining were evaluated once every two weeks. After four weeks of administration, mice were euthanized, and their lacrimal gland tissues and corneas were harvested. Hematoxylin-eosin (HE) staining was used to assess histopathological morphology. Western blot was performed to detect the protein expression levels of MEK, Ras, Raf, and ERK. Enzyme-linked immunosorbent assay (ELISA) was used to measure the contents and expressions of MEK, Ras, Raf, ERK, and interleukin (IL)-1β in lacrimal gland and corneal tissues of the mice in each group. Quantitative real-time polymerase chain reaction (Real-time PCR) was employed to determine mRNA expression levels of MEK, Ras, Raf, and ERK. ResultsThe Mimenghua prescription groups and the sodium hyaluronate group exhibited significantly increased tear secretion volume (P<0.05) and prolonged TBUT (P<0.05) after treatment. Ocular surface damage of mice was visibly recovered. Western blot results indicated that protein expression levels of MEK, Ras, Raf, and ERK in the lacrimal gland and corneal tissues were significantly downregulated in the sodium hyaluronate group and Mimenghua prescription group with high dose (P<0.05). ELISA results showed that IL-1β levels were highest in the model group but significantly reduced in the sodium hyaluronate group and Mimenghua prescription groups (P<0.05). Both ELISA and Real-time PCR results demonstrated that the expression levels of MEK, Ras, Raf, and ERK in the lacrimal glands and corneal tissues were significantly elevated in the model group (P<0.05), but markedly downregulated in the sodium hyaluronate group and Mimenghua prescription groups (P<0.05), suggesting that Mimenghua prescription can decrease the expressions of MEK, Ras, Raf, and ERK in the lacrimal glands and corneal tissues. ConclusionMimenghua prescription can reduce inflammatory responses, increase tear secretion, prolong TBUT, and promote corneal recovery by inhibiting the MEK, Ras, Raf, and ERK signaling pathways in lacrimal gland and corneal tissues.
4.Clinical effect of functional genomic analysis combined with individualized drug selection in treatment of autosomal dominant polycystic kidney disease with congenital hepatic fibrosis: A case report
Kaidi ZHU ; Jianzeng ZHANG ; Hongyi LI ; Mengqi YUAN ; Ziying ZHANG ; Zhe XU ; Hongling LIU ; Fusheng WANG ; Xuechun LU ; Lei SHI
Journal of Clinical Hepatology 2026;42(7):1670-1676
Autosomal dominant polycystic kidney disease (ADPKD) is a systemic hereditary renal disorder and can affect multiple organs, and congenital hepatic fibrosis is one of the manifestations of liver involvement and is an important complication of ADPKD. Symptomatic management is currently the main treatment method for this disease, and disease-specific drugs such as tolvaptan have limited indications and cannot correct the underlying genetic defect. This article reports a case of ADPKD with congenital hepatic fibrosis, and sirolimus was identified as the individualized treatment regimen based on peripheral blood functional genomic analysis and drug sensitivity prediction platform. The patient achieved significant improvements in symptoms and quality of life after treatment, with a stable kidney volume. This case shows that functional genomics has a potential value in guiding individualized treatment of rare genetic disorders, which provides new treatment ideas and practice paths for similar patients.
5.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.
6.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.
7.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.
8.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.
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 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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