1.Treatment Principles and Paradigm of Diabetic Microvascular Complications Responding Specifically to Traditional Chinese Medicine
Anzhu WANG ; Xing HANG ; Lili ZHANG ; Xiaorong ZHU ; Dantao PENG ; Ying FAN ; Min ZHANG ; Wenliang LYU ; Guoliang ZHANG ; Xiai WU ; Jia MI ; Jiaxing TIAN ; Wei ZHANG ; Han WANG ; Yuan XU ; .LI PINGPING ; Zhenyu WANG ; Ying ZHANG ; Dongmei SUN ; Yi HE ; Mei MO ; Xiaoxiao ZHANG ; Linhua ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):272-279
To explore the advantages of traditional Chinese medicine (TCM) and integrative TCM-Western medicine approaches in the treatment of diabetic microvascular complications (DMC), refine key pathophysiological insights and treatment principles, and promote academic innovation and strategic research planning in the prevention and treatment of DMC. The 38th session of the Expert Salon on Diseases Responding Specifically to Traditional Chinese Medicine, hosted by the China Association of Chinese Medicine, was held in Beijing, 2024. Experts in TCM, Western medicine, and interdisciplinary fields convened to conduct a systematic discussion on the pathogenesis, diagnostic and treatment challenges, and mechanism research related to DMC, ultimately forming a consensus on key directions. Four major research recommendations were proposed. The first is addressing clinical bottlenecks in the prevention and control of DMC by optimizing TCM-based evidence evaluation systems. The second is refining TCM core pathogenesis across DMC stages and establishing corresponding "disease-pattern-time" framework. The third is innovating mechanism research strategies to facilitate a shift from holistic regulation to targeted intervention in TCM. The fourth is advancing interdisciplinary collaboration to enhance the role of TCM in new drug development, research prioritization, and guideline formulation. TCM and integrative approaches offer distinct advantages in managing DMC. With a focus on the diseases responding specifically to TCM, strengthening evidence-based support and mechanism interpretation and promoting the integration of clinical care and research innovation will provide strong momentum for the modernization of TCM and the advancement of national health strategies.
2.Development and validation of an intelligent surveillance system for upper gastrointestinal high-risk patients
Mei DENG ; Guoen LYU ; Conghui SHI ; Jia LI ; Lianlian WU ; Jun LIU ; Honggang YU
Chinese Journal of Digestive Endoscopy 2025;42(3):190-196
Objective:To develop an intelligent surveillance system for identifying upper gastrointestinal high-risk patients and assigning surveillance intervals, and to verify its efficacy.Methods:The endoscopic and pathological reports of 23 035 patients undergoing endoscopy at Renmin Hospital of Wuhan University from January to October 2021 were collected retrospectively. A training set of 17 934 patients (January to August) and a test set of 5 101 patients (September to October) were established. Keywords in the endoscopic and pathological reports were extracted by the intelligent surveillance system, and high-risk patients were automatically identified and classified into 7 risk levels. Then the standardized surveillance intervals were assigned based on the guideline. Guideline-based surveillance intervals assigned by expert endoscopists based on endoscopic and pathological reports were used as the golden standard. The accuracy of the intelligent surveillance system was calculated. Of the patients within the test set, 189 were hospitalized and the surveillance intervals given by physicians could be obtained from the electronic health records. The accuracy of the intelligent surveillance system with that of physicians from different departments was compared. Then 67 patients were randomly selected from 189 patients by simple random sampling to evaluate the adjunctive effect of the system in assigning surveillance intervals among 3 endoscopists.Results:The overall accuracy of the intelligent surveillance system in identifying upper gastrointestinal high-risk patients was 99.94% (5 098/5 101), and that of assigning surveillance intervals to correctly included patients was 100.00% (534/534). The intelligent surveillance system achieved significantly higher accuracy compared with all physicians from different departments [98.94% (187/189) VS 35.45% (67/189), χ2=118.01, P<0.001] as well as physicians from department of gastroenterology [100.00% (117/117) VS 24.79% (29/117), χ2=86.01, P<0.001]. With the assistance of the intelligent surveillance system, the endoscopists' accuracy of assigning surveillance intervals to 67 patients was significantly improved [55.22% (111/201) VS 22.39% (45/201), χ2=58.68, P<0.001]. Conclusion:The intelligent surveillance system can accurately identify upper gastrointestinal high-risk patients and assign surveillance intervals according to risk levels, which can alleviate the workload of doctors and improve the follow-up rate of patients.
3.Mechanism of Zuoguiwan in Inhibiting Osteoclast Activation Induced by Breast Cancer via Regulating p38 MAPK/ERK Signaling Pathway
Jianjiang FU ; Yinlong MEI ; Junchao MA ; Xiaocui ZHU ; Wei WANG ; Hong LYU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):1-9
ObjectiveTo investigate the effects of Zuoguiwan on osteoclast activation induced by breast cancer and its mechanism. MethodsTo simulate breast cancer-induced osteoclastic bone metastasis, RAW264.7 cells were cultured in conditioned medium containing 50% supernatant of MDA-MB-231 breast cancer cells. The dosages of Zuoguiwan used in the experiment were sera containing 5% and 10% Zuoguiwan. Tartrate-resistant acid phosphatase (TRAP) staining was used to detect osteoclast activation. Enzyme-linked immunosorbent assay (ELISA) was used to measure Cathepsin K secretion from RAW264.7 cells. Real-time quantitative polymerase chain reaction (PCR) was used to detect the mRNA expression levels of osteocalcin (OCN) and bone sialoprotein (BSP). Immunoprecipitation was employed to detect the interaction between Runt-related transcription factor 2 (Runx2) and core binding factor β subunit (CBF-β). Western blot was used to assess the protein expression of Runx2, phosphorylated Runx2 (p-Runx2), extracellular signal-regulated kinases 1/2 (ERK1/2), p-ERK1/2, p38 mitogen-activated protein kinase (MAPK), p-p38 MAPK, and CBF-β. ResultsCompared with the blank group, the MDA-MB-231 cell supernatant group showed a significant increase in TRAP-positive cell counts and Cathepsin K secretion. Meanwhile, the expression levels of p-Runx2, Runx2-CBF-β interaction, BSP and OCN mRNA, p-p38 MAPK, and p-ERK1/2 proteins were significantly decreased (P<0.01). Compared with the MDA-MB-231 cell supernatant group, Zuoguiwan-containing sera significantly reduced TRAP-positive cell counts and Cathepsin K secretion (P<0.01), significantly increased p-Runx2, BSP and OCN mRNA expression, as well as p-p38 MAPK and p-ERK1/2 protein levels, and promoted the interaction between Runx2 and CBF-β (P<0.01). No significant change in Runx2 expression was observed. Compared to the blank group, the BVD-523 group showed significantly lower expression of p-p38 MAPK and p-ERK1/2 proteins (P<0.01). Compared with the BVD-523 group, both low and high concentration Zuoguiwan-containing sera groups showed significantly higher p-p38 MAPK expression (P<0.01), and the high concentration Zuoguiwan group also exhibited a significant increase in p-ERK1/2 expression (P<0.01), while no statistical difference was found in the low-dose group. ConclusionZuoguiwan inhibits osteoclast activation by inducing phosphorylation of the key transcriptional regulator Runx2 in intra-osteoclast bone formation, and this process is closely associated with the activation of the p38 MAPK/ERK signaling pathway.
4.Diagnostic value of cardiac color Doppler ultrasound for left ventricular hypertrophy complicated left heart failure
Qun-xing LYU ; Song-mei ZHANG ; Peng-sheng LIN ; Tan-shou CHEN
Chinese Journal of cardiovascular Rehabilitation Medicine 2025;34(2):184-189
Objective:To explore the diagnostic value of cardiac color Doppler ultrasonography for left ventricular hypertrophy(LVH)complicated left heart failure(LHF).Methods:A total of 117 LVH patients complicated LHF admitted in Ningde City Hospital Affiliated to Ningde Normal College between January 2019 and January 2023(heart failure group)and 100 healthy people(control group)who underwent general physical examination in our hospital simultaneously were selected for the study.Left ventricular ejection fraction(LVEF),left atrial diameter(LAD),left ventricular end-diastolic diameter(LVEDd),early diastolic peak flow velocity/late diastolic peak flow velocity(E/A)were measured by cardiac color Doppler ultrasound in two groups.Receiver operating charac-teristic(ROC)curve was used to analyze the diagnostic value of cardiac color Doppler ultrasound indexes for LVH complicated LHF.In addition,the 117 patients with LVH and LHF were divided into mild group(n=54)and moderate to severe group(n=63)according to cardiac function class.Spearman correlation analysis was used to an-alyze the associationof cardiac color ultrasound parameters with cardiac function class.Results:Compared with par-ticipants in control group,those in heart failure group had significant lower LVEF[(47.88±4.75)% vs.(69.81±5.64)%],and significant higher LAD[(44.03±4.88)mm vs.(27.56±2.76)mm vs.],LVEDd[(55.68±5.04)mm vs.(42.19±1.38)mm],E/A[(13.04±3.58)vs.(6.60±1.67)](P<0.001 all).ROC analysis indicated that the area under the curve(AUC)of the combination of various parameters of cardiac ultrasound for diagnosing LVH with LHF was significantly higher than those of the single tests(Combined detection:AUC=0.901,95%CI 0.853~0.937 vs LVEF:AUC=0.644,95%CI 0.577~0.708,LAD:AUC=0.703,95%CI 0.637~0.763,LVEDd:AUC=0.633,95%CI 0.565~0.697,E/A:AUC=0.748,95%CI 0.685~0.804,Z=7.062,5.764,7.292,4.864,P<0.001 all).Compared with patients in mild group,those in moderate to severe group had signifi-cant lower LVEF[(45.67±3.37)% vs.(50.47±4.86)%],and significant higher LAD[(46.31±4.42)mm vs.(41.36±3.98)mm],LVEDd[(60.09±1.75)mm vs.(50.53±1.41)mm]and E/A[(13.99±2.96)vs.(11.93±3.92)](P<0.01 all).Spearman correlation analysis indicated that LVEF was negatively correlated with cardiac function class(r=-0.474),while LAD(r=0.511),LVEDd(r=0.863),E/A(r=0.269)were positively corre-lated with cardiac function class(P<0.01 all).Conclusion:Cardiac color Doppler ultrasound could better diagnose LVH complicated LHF,and also could effectively predict the cardiac function of patients with LVH and LHF.
5.Efficacy of Guanxinning Tablets as an adjunctive treatment for coronary heart disease complicated by insomnia in older adult patients
Na HUANG ; Xiongsheng LYU ; Jianhua MEI
Chinese Journal of Primary Medicine and Pharmacy 2025;32(2):172-176
Objective:To investigate the efficacy of Guanxinning Tablets as an adjunctive treatment for coronary heart disease complicated by insomnia in older adult patients. Methods:A total of 80 patients with coronary heart disease complicated by insomnia, admitted to The Second People's Hospital of Lishui between December 2021 and December 2023, were selected for this study. A randomized, controlled trial design was used, with patients randomly assigned to either the control group or the observation group ( n = 40/group). The control group received conventional treatment along with basic sleep-promoting interventions, while the observation group received Guanxinning Tablets in addition to the treatment provided to the control group. The treatment duration for both groups was 8 weeks. The therapeutic effects, frequency and duration of angina attacks, Pittsburgh Sleep Quality Index score, Seattle Angina Questionnaire score, serum levels of interleukin-6, tumor necrosis factor-alpha, vascular endothelial growth factor, and the incidence of adverse reactions were compared between the two groups. Results:The clinical efficacy of the observation group was significantly better than that of the control group ( Z = 2.07, P < 0.05). After treatment, the frequency and duration of angina attacks in the observation group were (1.02 ± 0.31) times/d and (1.35 ± 0.27) min, both of which were significantly lower and shorter than those in the control group [(1.54 ± 0.40) times/d, (1.71 ± 0.36) min, t = -6.50, -5.06, both P < 0.05]. The Pittsburgh Sleep Quality Index score in the observation group was (5.93 ± 1.28), which was significantly lower than that in the control group [(7.33 ± 2.05), t = -3.66, P < 0.05]. The Seattle Angina Questionnaire scores for all dimensions in the observation group were higher than those in the control group ( t = 2.81, 2.30, 2.97, 4.76, 4.24, all P < 0.05). The levels of interleukin-6 and tumor necrosis factor-alpha in the observation group were (13.48 ± 3.60) mg/L and (15.53 ± 3.83) μg/L, respectively, both of which were significantly lower than those in the control group [(16.26 ± 4.51) mg/L, (20.38 ± 3.92) μg/L, t = -3.05, -5.60, both P < 0.05]. The vascular endothelial growth factor level in the observation group was (128.26 ± 16.67) ng/L, which was significantly higher than that in the control group [(105.78 ± 14.35) ng/L, t = 6.46, P < 0.05]. There was no significant difference in the incidence of adverse reactions between the two groups ( P > 0.05). Conclusions:Guanxinning Tablets, as an adjunctive treatment for coronary heart disease complicated by insomnia, can significantly improve outcomes in older adults by alleviating angina symptoms, enhancing sleep quality, and reducing inflammatory responses, without increasing the incidence of drug-related adverse reactions.
6.Diagnostic value of cardiac color Doppler ultrasound for left ventricular hypertrophy complicated left heart failure
Qun-xing LYU ; Song-mei ZHANG ; Peng-sheng LIN ; Tan-shou CHEN
Chinese Journal of cardiovascular Rehabilitation Medicine 2025;34(2):184-189
Objective:To explore the diagnostic value of cardiac color Doppler ultrasonography for left ventricular hypertrophy(LVH)complicated left heart failure(LHF).Methods:A total of 117 LVH patients complicated LHF admitted in Ningde City Hospital Affiliated to Ningde Normal College between January 2019 and January 2023(heart failure group)and 100 healthy people(control group)who underwent general physical examination in our hospital simultaneously were selected for the study.Left ventricular ejection fraction(LVEF),left atrial diameter(LAD),left ventricular end-diastolic diameter(LVEDd),early diastolic peak flow velocity/late diastolic peak flow velocity(E/A)were measured by cardiac color Doppler ultrasound in two groups.Receiver operating charac-teristic(ROC)curve was used to analyze the diagnostic value of cardiac color Doppler ultrasound indexes for LVH complicated LHF.In addition,the 117 patients with LVH and LHF were divided into mild group(n=54)and moderate to severe group(n=63)according to cardiac function class.Spearman correlation analysis was used to an-alyze the associationof cardiac color ultrasound parameters with cardiac function class.Results:Compared with par-ticipants in control group,those in heart failure group had significant lower LVEF[(47.88±4.75)% vs.(69.81±5.64)%],and significant higher LAD[(44.03±4.88)mm vs.(27.56±2.76)mm vs.],LVEDd[(55.68±5.04)mm vs.(42.19±1.38)mm],E/A[(13.04±3.58)vs.(6.60±1.67)](P<0.001 all).ROC analysis indicated that the area under the curve(AUC)of the combination of various parameters of cardiac ultrasound for diagnosing LVH with LHF was significantly higher than those of the single tests(Combined detection:AUC=0.901,95%CI 0.853~0.937 vs LVEF:AUC=0.644,95%CI 0.577~0.708,LAD:AUC=0.703,95%CI 0.637~0.763,LVEDd:AUC=0.633,95%CI 0.565~0.697,E/A:AUC=0.748,95%CI 0.685~0.804,Z=7.062,5.764,7.292,4.864,P<0.001 all).Compared with patients in mild group,those in moderate to severe group had signifi-cant lower LVEF[(45.67±3.37)% vs.(50.47±4.86)%],and significant higher LAD[(46.31±4.42)mm vs.(41.36±3.98)mm],LVEDd[(60.09±1.75)mm vs.(50.53±1.41)mm]and E/A[(13.99±2.96)vs.(11.93±3.92)](P<0.01 all).Spearman correlation analysis indicated that LVEF was negatively correlated with cardiac function class(r=-0.474),while LAD(r=0.511),LVEDd(r=0.863),E/A(r=0.269)were positively corre-lated with cardiac function class(P<0.01 all).Conclusion:Cardiac color Doppler ultrasound could better diagnose LVH complicated LHF,and also could effectively predict the cardiac function of patients with LVH and LHF.
7.Csde1 Mediates Neurogenesis via Post-transcriptional Regulation of the Cell Cycle.
Xiangbin JIA ; Wenqi XIE ; Bing DU ; Mei HE ; Jia CHEN ; Meilin CHEN ; Ge ZHANG ; Ke WANG ; Wanjing XU ; Yuxin LIAO ; Senwei TAN ; Yongqing LYU ; Bin YU ; Zihang ZHENG ; Xiaoyue SUN ; Yang LIAO ; Zhengmao HU ; Ling YUAN ; Jieqiong TAN ; Kun XIA ; Hui GUO
Neuroscience Bulletin 2025;41(11):1977-1990
Loss-of-function variants in CSDE1 have been strongly linked to neuropsychiatric disorders, yet the precise role of CSDE1 in neurogenesis remains elusive. In this study, we demonstrate that knockout of Csde1 during cortical development in mice results in impaired neural progenitor proliferation, leading to abnormal cortical lamination and embryonic lethality. Transcriptomic analysis revealed that Csde1 upregulates the transcription of genes involved in the cell cycle network. Applying a dual thymidine-labelling approach, we further revealed prolonged cell cycle durations of neuronal progenitors in Csde1-knockout mice, with a notable extension of the G1 phase. Intersection with CLIP-seq data demonstrated that Csde1 binds to the 3' untranslated region (UTR) of mRNA transcripts encoding cell cycle genes. Particularly, we uncovered that Csde1 directly binds to the 3' UTR of mRNA transcripts encoding Cdk6, a pivotal gene in regulating the transition from the G1 to S phases of the cell cycle, thereby maintaining its stability. Collectively, this study elucidates Csde1 as a novel regulator of Cdk6, sheds new light on its critical roles in orchestrating brain development, and underscores how mutations in Csde1 may contribute to the pathogenesis of neuropsychiatric disorders.
Animals
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Neurogenesis/genetics*
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Cell Cycle/genetics*
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Mice, Knockout
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Mice
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Neural Stem Cells/metabolism*
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DNA-Binding Proteins/metabolism*
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Cyclin-Dependent Kinase 6/genetics*
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Cell Proliferation
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3' Untranslated Regions
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Cerebral Cortex/embryology*
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RNA-Binding Proteins
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Mice, Inbred C57BL
8.Mechanistic of Yueju Wan volatile oil in inhibiting inflammation for antidepressant effects by regulating AGE/PI3K/Akt pathway.
Tan-Lu CHU ; Ze-Jun GUO ; Wei ZHANG ; Ling-Feng WANG ; Shu-Rui LYU ; Wan-Yu GUO ; Xiao-Ming ZHONG ; Feng-Mei QIU ; Zhen HUANG
China Journal of Chinese Materia Medica 2025;50(11):3147-3158
The antidepressant activity and molecular mechanisms of Yueju Wan volatile oil were investigated. The Yueju Wan volatile oil was extracted by using supercritical CO_2. Gas chromatography-mass spectrometry(GC-MS) combined with network pharmacology identified 28 chemical constituents in Yueju Wan volatile oil, primarily terpenes and lactones. A total of 123 overlapping targets were associated with depression, including core targets of interleukin-1β(IL-1β), signal transducer and activator of transcription 3(STAT3), and caspase-3(CASP3). These targets were mainly involved in the prolactin, advanced glycation end products/receptor(AGE/RAGE), and phosphoinositide 3-kinase/protein kinase B(PI3K/Akt) signaling pathways. A reserpine-induced depression mouse model was established to evaluate the therapeutic effects and mechanisms of Yueju Wan volatile oil. The effects of Yueju Wan volatile oil on depression-like behavior in mice were evaluated by analyzing body mass, body temperature index, tail suspension immobility time, forced swimming immobility time, and sucrose preference. Hematoxylin-eosin(HE) staining revealed neuronal protection of Yueju Wan volatile oil in the brain of mice. Enzyme-linked immunosorbent assay(ELISA) and Western blot were employed to detect the protein expression of AGEs, IL-1β, phosphorylated PI3K(p-PI3K), Akt, phosphorylated Akt(p-Akt), nuclear factor κB(NF-κB), and brain-derived neurotrophic factor(BDNF). Behavioral evaluation showed that Yueju Wan volatile oil could effectively control the decline of body mass and body temperature of depressed mice, reduce tail suspension and swimming immobility time, and enhance their preference for sucrose. Histopathological examination showed that Yueju Wan volatile oil could alleviate the neuronal damage in CA1 and dentate gyrus(DG) of the hippocampus of mice. ELISA and Western blot results showed that Yueju Wan volatile oil could significantly increase the protein expression levels of PI3K, Akt, and BDNF and significantly decrease the protein expression levels of AGEs, IL-1β, p-PI3K, p-Akt, and NF-κB in the hippocampus of mice. Furthermore, the p-PI3K/PI3K and p-Akt/Akt ratios were significantly decreased at medium and high doses. These findings suggest that the aromatherapy of Yueju Wan volatile oil can significantly improve reserpine-induced depression-like behavior in mice, which may be related to reducing the expression of neuronal membrane protein AGEs, reducing the phosphorylation levels of PI3K and Akt, inhibiting NF-κB entry into the nucleus, and alleviating the release of pro-inflammatory factors and nerve injury.
Animals
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Antidepressive Agents/chemistry*
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Mice
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Proto-Oncogene Proteins c-akt/immunology*
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Phosphatidylinositol 3-Kinases/immunology*
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Oils, Volatile/chemistry*
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Male
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Drugs, Chinese Herbal/chemistry*
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Signal Transduction/drug effects*
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Depression/metabolism*
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Glycation End Products, Advanced/immunology*
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Humans
9.Effects of ginsenoside Rb_1 on liver FXR pathway and liver and fecal bile acid profiles in rats induced by high-fat diet based on targeted metabolomics.
Xue LENG ; Yang LI ; Qi WANG ; Xin-Tong LI ; Mei-Jun LYU ; Yan-Na SUN
China Journal of Chinese Materia Medica 2025;50(16):4649-4658
A targeted metabolomics study was conducted on the bile acid profiles in the liver and feces of rats induced by a high-fat diet and intervened by ginsenoside Rb_1, along with the detection of FXR pathway gene expression in the liver, to explore and clarify its mechanism of action. The content of biochemical indicators in the serum were detected using an automatic biochemical analyzer. Hematoxylin and eosin(HE) staining and oil red O staining were used to detect pathological changes and lipid deposition in the liver. RT-PCR was used to detect the mRNA expression of FXR, small heterodimer partner(SHP), cholesterol 7 alpha-hydroxylase(CYP7A1), and sterol regulatory element-binding protein-1c(SREBP-1c) in the liver. Targeted bile acid metabolomics technology was employed to analyze changes in bile acid profiles in liver tissue and feces, and a correlation analysis was performed between key genes such as FXR, SHP, CYP7A1, SREBP-1c and differential bile acid metabolites. The results showed that ginsenoside Rb_1 significantly reduced the levels of total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), and high-density lipoprotein cholesterol(HDL-C) in the serum, alleviated the large fat vacuoles and lipid deposition in the liver, increased the expression of FXR mRNA in the liver, and decreased the expression of SREBP-1c mRNA. The expression of CYP7A1 and SHP mRNA was increased, but the differences were not statistically significant. Targeted bile acid metabolomics showed that ginsenoside Rb_1 could restore the levels of 9 bile acids in the liver and 8 bile acids in the feces. Ginsenoside Rb_1 also increased the percentage of taurocholic acid(TCA) in the liver(56.78%) and the percentage of 12-ketolithocholic acid(12-KLCA) in the feces(26.10%). Pathway enrichment analysis revealed two pathways involved in bile acid metabolism: primary bile acid biosynthesis and taurine and hypotaurine metabolism. Correlation analysis showed that FXR, SHP, CYP7A1, and SREBP-1c were positively correlated with multiple differential bile acids. These results suggest that ginsenoside Rb_1 may intervene in lipid metabolism disorders induced by a high-fat diet by regulating the FXR pathway and modulating bile acid profiles in the liver and feces.
Animals
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Bile Acids and Salts/metabolism*
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Rats
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Ginsenosides/pharmacology*
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Male
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Receptors, Cytoplasmic and Nuclear/genetics*
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Liver/drug effects*
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Diet, High-Fat/adverse effects*
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Metabolomics
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Rats, Sprague-Dawley
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Feces/chemistry*
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Cholesterol 7-alpha-Hydroxylase/metabolism*
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Sterol Regulatory Element Binding Protein 1/genetics*
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Humans
10.Mechanism of Liangfang Wenjing Decoction in Regulating Endoplasmic Reticulum Stress to Reduce Apoptosis and Alleviate Ovarian Microvascular Injury in Rats with Cold Coagulation and Blood Stasis Syndrome
Tianyuan LYU ; Xueyan MA ; Yue HU ; Liqun FENG ; Xiaodan SONG ; Lianmin MEI ; Xiumei CHENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):103-114
ObjectiveTo observe ovarian microvascular damage in rats with cold coagulation and blood stasis syndrome and to explore the mechanism by which Liangfang Wenjing decoction improves this condition in rats. MethodsFifty SPF female SD rats were randomly divided into a blank group, a model group, low-dose (8.1 g·kg-1) and high-dose groups (16.2 g·kg-1) of Liangfang Wenjing decoction, and a 4-phenylbutyric acid (0.1 g·kg-1) group, with 10 rats in each group. The ice-water bath method was employed to establish the rat model of cold coagulation and blood stasis syndrome. Concurrent with modeling, Liangfang Wenjing decoction was administered continuously for 21 days, once daily. The rats' syndrome manifestations and estrous cycles were recorded. The enzyme-linked immunosorbent assay (ELISA) was used to detect serum reproductive hormone levels and levels of endothelin-1 (ET-1), nitric oxide (NO), thrombomodulin (TM), and von Willebrand factor (vWF) in ovarian tissue. Prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), and fibrinogen (FIB) were measured. The ovarian microcirculatory blood perfusion was detected by laser speckle contrast imaging. Hematoxylin-eosin (HE) staining was performed to observe the ovarian histopathology, flow cytometry to detect ovarian apoptosis rate, and transmission electron microscopy to observe the ultrastructure of ovarian microvascular endothelial cells. Western blot was employed to detect the protein expression of endothelial nitric oxide synthase (eNOS), phosphorylated eNOS (p-eNOS), Caspase-3, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), glucose-regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), inositol-requiring enzyme1α (IRE1α), p-IRE1α, apoptosis signal-regulating kinase 1 (ASK1), p-ASK1, c-Jun N-terminal kinase (JNK), and p-JNK. Immunofluorescence was used to detect ovarian Bax and Bcl-2 expression in microvascular endothelial cells. ResultsCompared with the blank group, the model group showed signs of cold coagulation and blood stasis syndrome, prolonged estrus cycles, and reproductive hormone disorders. Histopathological results revealed a decrease in follicle counts at all stages and disorganized granulosa cell arrangement. Ovarian microcirculatory perfusion was significantly decreased (P<0.01). PT, APTT, and TT were reduced (P<0.05, P<0.01), while FIB levels were increased (P<0.05). In ovarian tissue, NO content was decreased, while ET-1, vWF, and TM levels were increased significantly (P<0.01). The apoptosis rate of ovarian cells was markedly increased (P<0.01). Furthermore, p-eNOS/eNOS and Bcl-2 were decreased (P<0.05), whereas Bax, cleaved-Caspase-3/Caspase-3, GRP78, CHOP, p-IRE1α/IRE1α, p-ASK1/ASK1, and p-JNK/JNK expression showed significant increases (P<0.05, P<0.01). Compared with the model group, Liangfang Wenjing decoction intervention alleviated the symptoms of cold coagulation and blood stasis, gradually restored the estrus cycle, and improved ovarian histopathology and endothelial cell ultrastructure. Microcirculatory blood perfusion was significantly elevated (P<0.05). NO content in ovarian tissue was elevated, while ET-1, vWF, and TM levels were significantly decreased (P<0.05, P<0.01). The p-eNOS/eNOS ratio and Bcl-2 expression were significantly elevated (P<0.05), while the expression of Bax, cleaved-Caspase-3/Caspase-3, GRP78, CHOP, p-IRE1α/IRE1α, p-ASK1/ASK1, and p-JNK/JNK was significantly decreased (P<0.05, P<0.01). ConclusionLiangfang Wenjing decoction may regulate the IRE1α/ASK1/JNK signaling pathway to inhibit endoplasmic reticulum stress, attenuate apoptosis, and improve microvascular endothelial injury in ovaries of rats with cold coagulation and blood stasis syndrome.

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