1.Bioinformatics Reveals Mechanism of Xiezhuo Jiedu Precription in Treatment of Ulcerative Colitis by Regulating Autophagy
Xin KANG ; Chaodi SUN ; Jianping LIU ; Jie REN ; Mingmin DU ; Yuan ZHAO ; Xiaomeng LANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):166-173
ObjectiveTo explore the potential mechanism of Xiezhuo Jiedu prescription in regulating autophagy in the treatment of ulcerative colitis (UC) by bioinformatics and animal experiments. MethodsThe differentially expressed genes (DEGs) in the colonic mucosal tissue of UC patients was obtained from the Gene Expression Omnibus (GEO), and those overlapped with autophagy genes were obtained as the differentially expressed autophagy-related genes (DEARGs). DEARGs were imported into Metascape and STRING, respectively, for gene ontology/Kyoto Encyclopedia of Genes and Genomics (GO/KEGG) enrichment analysis and protein-protein interaction (PPI) analysis. Finally, 15 key DEARGs were obtained. The core DEARGs were obtained by least absolute shrinkage and selection operator (LASSO) regression and receiver operating characteristic curve (ROC) analysis. The CIBERSORT deconvolution algorithm was used to analyze the immunoinfiltration of UC patients and the correlations between core DEARGs and immune cells. C57BL/6J mice were assigned into a normal group and a modeling group. The mouse model of UC was established by free drinking of 2.5% dextran sulfate sodium. The modeled mice were assigned into low-, medium-, and high-dose Xiezhuo Jiedu prescription and mesalazine groups according to the random number table method and administrated with corresponding agents by gavage for 7 days. The colonic mucosal morphology was observed by hematoxylin-eosin staining. The protein and mRNA levels of cysteinyl aspartate-specific proteinase 1 (Caspase-1), cathepsin B (CTSB), C-C motif chemokine-2 (CCL2), CXC motif receptor 4 (CXCR4), and hypoxia-inducing factor-1α (HIF-1α) in the colon tissue were determined by Western blot and real-time fluorescence quantitative polymerase chain reaction, respectively. ResultsThe dataset GSE87466 was screened from GEO and interlaced with autophagy genes. After PPI analysis, LASSO regression, and ROC analysis, the core DEARGs (Caspase-1, CCL2, CTSB, and CXCR4) were obtained. The results of immunoinfiltration analysis showed that the counts of NK cells, M0 macrophages, M1 macrophages, and dendritic cells in the colonic mucosal tissue of UC patients had significant differences, and core DEARGs had significant correlations with these immune cells. This result, combined with the prediction results of network pharmacology, suggested that the HIF-1α signaling pathway may play a key role in the regulation of UC by Xiezhuo Jiedu prescription. The animal experiments showed that Xiezhuo Jiedu prescription significantly alleviated colonic mucosal inflammation in UC mice. Compared with the normal group, the model group showed up-regulated protein and mRNA levels of caspase-1, CCL2, CTSB, CXCR4, and HIF-1α, which were down-regulated after treatment with Xiezhuo Jiedu prescription or mesalazine. ConclusionCaspase-1, CCL2, CTSB, and CXCR4 are autophagy genes that are closely related to the onset of UC. Xiezhuo Jiedu prescription can down-regulate the expression of core autophagy genes to alleviate the inflammation in the colonic mucosa of mice.
2.Effect of Yiqi Wenyang Huoxue Lishui Components on Cardiac Function and Mitochondrial Energy Metabolism in CHF Rats
Hui GAO ; Zeqi YANG ; Xin LIU ; Fan GAO ; Yangyang HAN ; Aiyangzi LU ; Xingchao LIU ; Qiuhong GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):27-36
ObjectiveTo investigate the effects of Yiqi Wenyang Huoxue Lishui components on the cardiac function and mitochondrial energy metabolism in the rat model of chronic heart failure (CHF) and explore the underlying mechanism. MethodsThe rat model of CHF was prepared by transverse aortic constriction (TAC). Eight of the 50 SD rats were randomly selected as the sham group, and the remaining 42 underwent TAC surgery. The 24 SD rats successfully modeled were randomized into model, trimetazidine (6.3 mg·kg-1), and Yiqi Wenyang Huoxue Lishui components (60 mg·kg-1 total saponins of Astragali Radix, 10 mg·kg-1 total phenolic acids of Salviae Miltiorrhizae Radix et Rhizoma, 190 mg·kg-1 aqueous extract of Lepidii Semen, and 100 mg·kg-1 cinnamaldehyde) groups. The rats were administrated with corresponding agents by gavage, and those in the sham and model groups were administrated with the same amount of normal saline at a dose of 10 mL·kg-1 for 8 weeks. Echocardiography was used to examine the cardiac function in rats. Enzyme-linked immunosorbent assay was employed to determine the serum levels of N-terminal pro-B-type natriuretic peptide (NT-ProBNP), hypersensitive troponin(cTnI), creatine kinase (CK), lactate dehydrogenase (LD), free fatty acids (FFA), superoxide dismutase (SOD), and malondialdehyde (MDA). The colorimetric assay was employed to measure the levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) in the myocardial tissue. The pathological changes in the myocardial tissue were observed by hematoxylin-eosin staining and Masson staining. The Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities in the myocardial tissue were determined by the colorimetric assay. The ultrastructural changes of myocardial mitochondria were observed by transmission electron microscopy. Western blot was employed to determine the protein levels of ATP synthase subunit delta (ATP5D), glucose transporter 4 (GLUT4), and carnitine palmitoyltransferase-1 (CPT-1). The mitochondrial complex assay kits were used to determine the activities of mitochondrial complexes Ⅰ, Ⅱ, Ⅲ, and Ⅳ. ResultsCompared with the sham group, the model group showed a loosening arrangement of cardiac fibers, fracture and necrosis of partial cardiac fibers, inflammatory cells in necrotic areas, massive blue fibrotic tissue in the myocardial interstitium, increased collagen fiber area and myocardial fibrosis, destroyed mitochondria, myofibril disarrangement, sparse myofilaments, and fractured and reduced cristae. In addition, the rats in the model group showed declined ejection fraction (EF) and fractional shortening (FS), risen left ventricular end-diastolic diameter (LVIDd), left ventricular end-systolic diameter (LVIDs), left ventricular end-diastolic posterior wall thickness (LVPWd), left ventricular end-systolic posterior wall thickness (LVPWs), left ventricular end-diastolic volume (LVVOLd), and left ventricular end-systolic volume (LVVOLs), elevated levels of NT-ProBNP, cTnI, CK, MDA, FFA, and LD, lowered level of SOD, down-regulated protein levels of GLUT4 and CPT-1, decreased activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase, and respiratory complexes Ⅰ-Ⅳ, and declined levels of ATP5D, ATP, ADP, and AMP (P<0.05, P<0.01). Compared with the model group, the Yiqi Wenyang Huoxue Lishui components and trimetazidine groups showed alleviated pathological damage of the mitochondria and mycardial tissue, risen EF and FS, declined LVIDd, LVIDs, LVPWd, LVPWs, LVVOLd, and LVVOLs, lowered levels of NT-ProBNP, cTnI, CK, MDA, FFA, and LD, elevated level of SOD, up-regulated protein levels of GLUT4 and CPT-1, increased activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase, and respiratory complexes Ⅰ-Ⅳ, and elevated levels of ATP5D, ATP, ADP, and AMP (P<0.05, P<0.01). ConclusionYiqi Wenyang Huoxue Lishui components can improve the cardiac function, reduce myocardial injury, regulate glucose and lipid metabolism, optimize the utilization of substrates, and alleviate the damage of mitochondrial structure and function, thus improving the energy metabolism of the myocardium in the rat model of CHF.
3.Effect of "Fahan" on Metabolites of Blumea balsamifera Analyzed by Non-targeted Metabolomics
Jiayuan CAO ; Xin XU ; Xiangsheng ZHANG ; Bingnan LIU ; Yongyao WEI ; Ke ZHONG ; Yuxin PANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):200-207
ObjectiveTo characterize the changes of metabolites of Blumea balsamifera in the process of sweating by non-targeted metabolomics, and to investigate the influence of sweating processing on the constituents of B. balsamifera. MethodsUltra performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Exactive Orbitrap-MS) metabolomics was used to identify the metabolites in no sweating group(F1), sweating 2 d group(F2) and sweating 4 d group(F3), the differences of metabolites between the groups were compared by principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA), and differential metabolites were screened according to the variable importance in the projection(VIP) value>1 and P<0.05, and the pathway enrichment of the differential metabolites was analyzed by Kyoto Encyclopedia of Genes and Genomes(KEGG). ResultsThe results of PCA and OPLS-DA showed a clear distinction between the three groups of samples, indicating significant differences in the compositions of the three groups of samples. A total of 433 differential metabolites were screened between the F1 and F2, with 154 up-regulated and 279 down-regulated, the significant up-regulated metabolites were tangeritin, 5-O-demethylnobiletin and so on, while the metabolites with significant down-regulation included alternariol, fortunellin, etc. A total of 379 differential metabolites were screened between the F2 and F3, with 150 up-regulated and 229 down-regulated, the significant up-regulated metabolites were isoimperatorin, helianyl octanoate and so on, and the significant down-regulated metabolites were hovenoside I, goyasaponin Ⅲ, etc. KEGG pathway enrichment analysis showed that tyrosine metabolism, isoquinoline alkaloid biosynthesis, phenylalanine, tyrosine and tryptophan biosynthesis, tryptophan metabolism, valine, leucine and isoleucine biosynthesis, pantothenate and coenzyme A biosynthesis may be the key pathways affecting metabolite differences of B. balsamifera after sweating treatment. ConclusionSweating can reduce the content of endophytic mycotoxins in B. balsamifera and has a great impact on the synthesis and metabolic pathways of total flavonoids and auxin. This study can provide a reference for the process research on the sweating conditions of B. balsamifera.
4.Yishen Huashi Granules Protect Kidneys of db/db Mice via p38 MAPK Signaling Pathway
Kaidong ZHOU ; Sitong WANG ; Ge JIN ; Yanmo CAI ; Xin ZHOU ; Yunhua LIU ; Xinxue ZHANG ; Min ZHANG ; Zongjiang ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):58-68
ObjectiveTo explore the mechanism of Yishen Huashi granules in alleviating renal tubular epithelial cell injury and relieving diabetic kidney disease by regulating the mitogen-activated protein kinase (MAPK) signaling pathway. MethodsThe db/db mice of 12 weeks old were randomly assigned into model , dapagliflozin (1.6 mg·kg-1), and Yishen Huashi granules (4.7 g·kg-1), and db/m mice were used as the control group. The general conditions of mice were observed, and fasting blood glucose and 24-h urinary protein and albumin-to-creatinine ratio (ACR) were measured at weeks 0 and 12 of administration. After 12 weeks of treatment, the levels of serum creatinine (SCr), blood urea (UREA), triglycerides (TG), total cholesterol (TC), and low density lipoprotein (LDL) were measured. The pathological changes in the renal tissue were observed by hematoxylin-eosin (HE) staining, Periodic acid-Schiff (PAS) staining, Mallory staining, and transmission electron microscopy. Real-time PCR was employed to determine the mRNA levels of monocyte chemotactic protein-1 (MCP-1) and CC chemokine receptor-2 (CCR2) in the renal tissue of mice. The immunohistochemical assay was employed to examine the expression of p38, phospho-p38 (p-p38), MCP-1, and CCR2 in the renal tissue of mice. Western blotting was employed to measure the protein levels of p-p38, p38, MCP-1, and CCR2 in the renal tissue of mice.HK-2 cells cultured in vitro were grouped as follows: negative control, high glucose(30 mmol·L-1), Yishen Huashi granule-containing serum, and SB203580. After 48 h of cell culture in each group, RNA were extracted and the levels of MCP-1, and CCR2 mRNA were determined by Real-time PCR,proteins were extracted and the levels of p38, p-p38, MCP-1, and CCR2 were determined by Western blot. ResultsThe in vivo experiments showed that before treatment, other groups had higher body weight, blood glucose level, 24 h urinary protein, and ACR than the control group (P<0.05,P<0.01). After 12 weeks of treatment, compared with the model group, the Yishen Huashi granules group showed improved general conditions, a decreasing trend in body weight, lowered levels of blood glucose, 24-h urinary protein, and ACR (P<0.01), reduced SCr and UREA (P<0.01), and declined levels of TC, TG, and LDL (P<0.05,P<0.01). Compared with the model group, the Yishen Huashi granules group showed alleviated damage and interstitial fibrosis in the renal tissue as well as reductions in glomerular foot process fusion and basement membrane thickening. Moreover, the Yishen Huashi granules group showed down-regulated mRNA levels of MCP-1 and CCR2 (P<0.01), reduced positive expression of p-p38, MCP-1, and CCR2 (P<0.01), and down-regulated protein levels of p-p38/p38, MCP-1, and CCR2 (P<0.05) in the renal tissue. The cell experiment showed that compared with the high glucose group, the Yishen Huashi granule-containing serum group showcased down-regulated mRNA levels of MCP-1 and CCR2 (P<0.01) and down-regulated protein levels of p-p38/p38, MCP-1, and CCR2(P<0.05,P<0.01). ConclusionYishen Huashi granules can regulate glucose-lipid metabolism, reduce 24 h urinary protein and ACR, improve the renal function, alleviate the renal tubule injury caused by high glucose, and protect renal tubule epithelial cells in db/db mice by reducing MCP-1/CCR2 activation via the p38 MAPK signaling pathway.
5.Analysis and evaluation of platelet bank establishment strategy from the perspective of donor loss
Zheng LIU ; Yamin SUN ; Xin PENG ; Yiqing KANG ; Ziqing WANG ; Jintong ZHU ; Juan DU ; Jianbin LI
Chinese Journal of Blood Transfusion 2025;38(2):238-243
[Objective] To analyze the loss rate of platelet donors and evaluate the strategies for establishing a platelet donor bank. [Methods] A total of 1 443 donors who joined the HLA and HPA gene donor bank for platelets in Henan Province from 2018 to 2020 were included in this study. Data on the total number of apheresis platelet donations, annual donation frequency, age at enrollment, donation habits (including the number of platelets donated per session and whether they had previously donated whole blood), and enrollment location were collected from the platelet donor information management system. Donor loss was determined based on the date of their last donation. The loss rates of different groups under various conditions were compared to assess the enrollment strategies. [Results] By the time the platelet bank was officially operational in 2022, 421 donors had been lost, resulting in an loss rate of 29% (421/1 443). By the end of 2023, the overall cumulative loss rate reached 52% (746/1 443). The loss rate was lower than the overall level in groups meeting any of the following conditions: total apheresis platelet donations exceeding 50, annual donation frequency of 10 or more, age at enrollment of 40 years or older, donation of more than a single therapeutic dose per session, or a history of whole blood donation two or more times. Additionally, loss rates varied across different enrollment locations, with higher enrollment numbers generally associated with higher loss rates. [Conclusion] Through a comprehensive analysis of donor loss, our center has adjusted its strategies for establishing the donor pool. These findings also provide valuable insights for other blood collection and supply institutions in building platelet donor banks.
6.Mechanism of Modified Danggui Shaoyaosan in Improving Inflammation and Apoptosis in Acne via Regulating JNK/p38 MAPK Pathway
Gongzhen CHEN ; Yuqi YANG ; Xin LIU ; Ting TANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):31-40
ObjectiveTo explore the therapeutic effects and mechanisms of modified Danggui Shaoyaosan on acne based on the c-Jun N-terminal kinase (JNK)/p38 mitogen-activated protein kinase (p38 MAPK) pathway. MethodsA rat ear acne model was established in SD rats, and the rats were divided into a blank group, a model group, and low-, medium-, and high-dose groups of modified Danggui Shaoyaosan (7.15, 14.30, 28.60 g·kg·d-1), with six rats in each group. After the administration for 14 consecutive days, morphological changes in the rats' auricles were observed, and hematoxylin-eosin (HE) staining was used to examine the pathological changes in the acne-affected ear tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the ear tissue. Quantitative reverse transcription polymerase chain reaction (Real-time PCR) was performed to detect the mRNA expression levels of Caspase-3, B cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), JNK, and p38 MAPK in the ear tissue. Additionally, Western blot analysis was conducted to assess the protein levels of Caspase-3, Bcl-2, Bax, JNK, and p38 MAPK in the ear tissue. The active components and key targets of modified Danggui Shaoyaosan in the treatment of acne were identified through network pharmacology analysis. Molecular docking was then employed to evaluate the interactions between the main active components and the key targets. ResultsThe results of the animal experiment demonstrated that compared with those in the blank group, rats in the model group exhibited redness, swelling, thickening, hardening, dryness, and roughness of the auricle. The surface showed sebaceous scales and desquamation, accompanied by acne-like lesions such as papule-like elevations or cysts. Histopathological changes included keratinization, epidermal thickening, dermal collagen fiber degeneration and necrosis, subcutaneous muscle degeneration and necrosis, inflammatory cell infiltration, and fibrous tissue proliferation. The mRNA and protein expression levels of IL-1β, TNF-α, Caspase-3, Bax, JNK, and p38 MAPK were significantly increased (P<0.01), while those of Bcl-2 were significantly decreased (P<0.01). In comparison to the model group, the modified Danggui Shaoyaosan groups showed marked improvement in acne-like lesions of the auricle, with varying degrees of histopathological damage reduction. Additionally, the mRNA and protein expression levels of IL-1β, TNF-α, Caspase-3, Bax, JNK, and p38 MAPK in the tissue were significantly decreased (P<0.05, P<0.01), while those of Bcl-2 were significantly increased (P<0.05, P<0.01). Network pharmacology analysis indicated that the key compounds in modified Danggui Shaoyaosan responsible for its effects in treating acne may include acacetin, kaempferol, luteolin, quercetin, wogonin, and baicalein. These compounds exerted their effects by modulating core targets such as TNF, IL-1β, Caspase-3, and Bcl-2, thereby alleviating inflammation and apoptosis and ultimately improving acne symptoms. ConclusionModified Danggui Shaoyaosan may exert its therapeutic effects on acne by inhibiting the activation of the JNK/p38 MAPK pathway, thereby alleviating inflammation and apoptosis.
7.Clinical Diseases Responding Specially to TCM Treatment: Psoriasis
Liu LIU ; Xiaoying SUN ; Mei MO ; Yaqiong ZHOU ; Bin LI ; Xiaoxiao ZHANG ; Xin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):260-268
Psoriasis is a common chronic inflammatory systemic disease in dermatology. Its high prevalence, recurrence rate, and numerous comorbidities impose a significant physical and mental burden on patients. With the continuous advancement of modern medicine, the emergence of biological agents has improved clinical efficacy, making it possible to overcome psoriasis, in addition to classical treatments. However, in clinical practice, adverse reactions, drug resistance, recurrence rates, and immune drift cannot be ignored. Traditional Chinese medicine (TCM) has a history of thousands of years in treating psoriasis, demonstrating good efficacy, high safety, and a low recurrence rate, but a standardized management system is lacking. Therefore, the 25th Clinical Diseases Responding Specially to TCM Treatment Series (Psoriasis) Youth Salon, hosted by the Chinese Association of Chinese Medicine and organized by the Youth Committee of the Chinese Association of Chinese Medicine, invited 29 experts and scholars from TCM, Western medicine, and interdisciplinary fields to actively discuss the "Advantages, Challenges, and Clinical Transformation of TCM and Western Medicine in the Diagnosis and Treatment of Psoriasis". The experts at the meeting concluded that the advantages of TCM in the treatment of psoriasis are as follows. Firstly, in the TCM-led treatment plan, TCM's understanding of psoriasis follows the principle of combining the differentiation of disease and syndrome. This approach distinguishes the basic contradiction from the current main contradiction and enables a clear grasp of the dynamic process of psoriasis development. Based on the system of syndrome differentiation and treatment, TCM intervention is applied to address the current main contradiction, and the optimal TCM treatment plan is formulated by combining internal and external treatments. Adhering to the principle of "what is visible outside must be addressed inside", TCM can prevent and treat psoriasis comorbidities early by differentiating syndrome types. Secondly, in the integrated TCM and Western medicine treatment plan, the combination of both methods not only enhances efficacy but also reduces the adverse reactions of immunosuppressants and biological agents, lowering the recurrence rate. This conference provides a reference for the diagnosis and treatment of psoriasis using TCM and integrated TCM and Western medicine, opening up new ideas for clinical and basic research and guiding future research directions.
8.Hepatitis E virus infection among blood donors in Ningbo
Mingxi PENG ; Yiyu LIU ; Huyan MAO ; Dan LIN ; Lu XIN ; Ning SHU ; Jianfeng HAN ; Feng DING
Chinese Journal of Blood Transfusion 2025;38(1):7-12
[Objective] To investigate the infection status and characteristics of HEV among voluntary blood donors in Ningbo, and to provide a basis for improving the blood screening strategy. [Methods] A total of 12 227 blood samples from voluntary blood donors in Ningbo from June 2022 to May 2023 were tested for HEV serology, enzymology, and nucleic acid testing. Furthermore, HEV gene sequencing was performed for genotyping analysis, and donors with reactive nucleic acid testing results were followed up to confirm their infection status. [Results] The reactivity rate of HEV Ag, anti-HEV IgM and anti-HEV IgG was 0.098%, 0.899% and 29.198%, respectively. There was no difference in the reactivity of anti-HEV IgM and anti-HEV IgG between genders, donation frequencies and donation types (P>0.05). The reactivity rate increased significantly with age (P<0.05). The rate of ALT disqualification (ALT>50U/L) was significantly higher than that in non-reactive samples (P<0.05). The HEV Ag reactivity rate (0.098%) was not correlated with gender, donation frequency, donation type or age. One HEV RNA positive case was found, with a positive rate of 0.008%(1/12 227). It was confirmed to be hepatitis E virus genotype 3 by sequencing analysis. Apart from HEV Ag reactivity, all other blood safety screening items were non-reactive, suggesting this case might be in the acute infection phase. The follow-up results showed that all indicators of the donor's previous blood donation were non-reactive. [Conclusion] Pre-donation ALT detection can reduce the risk of transfusion-transmitted HEV (TT-HEV) to a certain extent, and the effective way to prevent TT-HEV is to detect HEV RNA and serology of donor blood.
9.Molecular biological research and molecular homologous modeling of Bw.03 subgroup
Li WANG ; Yongkui KONG ; Huifang JIN ; Xin LIU ; Ying XIE ; Xue LIU ; Yanli CHANG ; Yafang WANG ; Shumiao YANG ; Di ZHU ; Qiankun YANG
Chinese Journal of Blood Transfusion 2025;38(1):112-115
[Objective] To study the molecular biological mechanism for a case of ABO blood group B subtype, and perform three-dimensional modeling of the mutant enzyme. [Methods] The ABO phenotype was identified by the tube method and microcolumn gel method; the ABO gene of the proband was detected by sequence-specific primer polymerase chain reaction (PCR-SSP), and the exon 6 and 7 of the ABO gene were sequenced and analyzed. Homologous modeling of Bw.03 glycosyltransferase (GT) was carried out by Modeller and analyzed by PyMOL2.5.0 software. [Results] The weakening B antigen was detected in the proband sample by forward typing, and anti-B antibody was detected by reverse typing. PCR-SSP detection showed B, O gene, and the sequencing results showed c.721 C>T mutation in exon 7 of the B gene, resulting in p. Arg 241 Trp. Compared with the wild type, the structure of Bw.03GT was partially changed, and the intermolecular force analysis showed that the original three hydrogen bonds at 241 position disappeared. [Conclusion] Blood group molecular biology examination is helpful for the accurate identification of ambiguous blood group. Homologous modeling more intuitively shows the key site for the weakening of Bw.03 GT activity. The intermolecular force analysis can explain the root cause of enzyme activity weakening.
10.Mechanism of Xinnao shutong capsule alleviating cerebral ischemia-reperfusion injury in rats by regulating ferroptosis
Huani LI ; Changhe LIU ; Xiaoyan GUO ; Xin ZHONG ; Wei ZHANG ; Wenjing GE
China Pharmacy 2025;36(3):306-311
OBJECTIVE To study the mechanism of Xinnao shutong capsule alleviating cerebral ischemia reperfusion injury (CIRI) in rats by regulating the ferroptosis pathway. METHODS SD rats were randomly divided into sham operation group, model group, Xinnao shutong low-dose, high-dose group (220, 440 mg/kg), Ginkgo biloba leaves extract group (positive control, 150 mg/kg). Each group of rats was orally administered with the corresponding medication/normal saline for 7 consecutive days. Transient occlusion of the middle cerebral artery was adopted to induce the CIRI model; the samples were taken 24 h after the operation; the cerebral infarction area of rats was detected, and the cerebral infarction rate was calculated. The pathological changes of brain tissues were observed, and the levels of lipid peroxide (LPO), malondialdehyde (MDA) and glutathione (GSH) in cerebral tissue were detected; mRNA and protein expressions of nuclear factor-erythroid 2-related factor 2 (Nrf2), heme oxygenase 1(HO-1) and glutathione peroxidase 4 (GPX4) were all detected in cerebral tissue of rats. RESULTS Compared with model group, the cerebral infarction rate, the content of total iron in cerebral tissue and serum level of LPO (except for Ginkgo biloba leaves extract group and Xinnao shutong low-dose group) were all decreased significantly in G. biloba leaves extract group and Xinnao shutong groups (P<0.05 or P<0.01); the serum level of GSH, the protein and mRNA expressions of Nrf2, HO-1 and GPX4 were all increased significantly (P<0.05 or P<0.01). The pathological damage to brain tissue was reduced, the number of nerve cells increased, the edema was alleviated, and the nuclear membrane was flattened. CONCLUSIONS Xinnao shutong capsule can inhibit ferroptosis and reduce CIRI, the mechanism of which may be associated with the activation of the Nrf2/HO-1/GPX4 signaling pathway.

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