1.Analysis of Blood-absorbed Components and Their Metabolic Differences of Xiebaisan in Normal and Chronic Bronchitis Mice Based on UPLC-Q-Exactive Orbitrap MS
Peng PENG ; Jiaxin LI ; Xinyue YANG ; Fangle LIU ; Chenchen ZHU ; Chaozhan LIN ; Yufeng YAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):219-227
ObjectiveThis study aims to systematically analyze the blood-absorbed components and metabolic profiles of Xiebaisan(XBS) in normal and chronic bronchitis (CB) mice using ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS), while comparing differences between the two states. MethodsThirty female BABL/c mice were randomly divided into the normal group, the normal drug administration group, the CB group, the CB drug administration group and the dexamethasone group, with 6 mice in each group. The CB mouse model was established by inducing with ovalbumin (OVA). The mice in the normal drug administration group and the CB drug administration group started to be gavaged with XBS(13.2 g·kg-1) from the 21st day, and the dexamethasone group mice were simultaneously gavaged with dexamethasone (0.5 mg·kg-1) until the end of the 35th day of the experiment. Subsequently, serum samples were collected and evaluated for their efficacy, based on the pharmacological evaluation indicators, to determine the efficacy of XBS in treating CB. Then the UPLC-Q-Exactive Orbitrap MS was employed to identify and analyze the chemical constituents, blood-absorbed components, and metabolites of XBS. Chemometric analysis was conducted to reveal metabolic profile differences under "dual states". Concurrently, Real-time PCR technology was utilized to detect the expression levels of key liver metabolic enzymes CYP2E1, CYP3A1, UGT1A1, and UGT1A6. ResultsA total of 28 prototype components and 158 metabolites (including 48 phase Ⅰ metabolites and 110 phase Ⅱ metabolites) of XBS were unambiguously identified in the serum of normal mice. Additionally, a comprehensive characterization was performed on a total of 32 prototype components and 178 metabolites (including 50 phase Ⅰ metabolites and 128 phase Ⅱ metabolites) of XBS in the serum of CB mice. Among them, 27 prototype components were detected in both states, including 12 flavonoids, 2 alkaloids, 3 triterpenes, 4 organic acids, 3 amides, 1 stilbene and 2 other compounds. The chemometrics analysis revealed no significant difference in the prototype components and metabolites of XBS between normal and CB mice; however, there was a significant increase in the in-vivo exposure of XBS in CB mice. Compared to normal mice, the levels of phase Ⅰ metabolites such as oxidation, reduction and methylation of blood components of XBS as well as phase Ⅱ metabolites of glucuronidation showed significant changes in CB mice. Real-time PCR further confirmed that these alterations were attributed to the upregulation of CYP2E1 (P<0.05), CYP3A1 (P>0.05), UGT1A1 (P<0.01) and UGT1A6 (P<0.01) enzymes expression in the liver of CB mice. ConclusionThis study elucidated the disparities in the levels of the blood-absorbed components and metabolic profiles of XBS in normal and CB mice, especially in oxidation, reduction, methylation in phase Ⅰ metabolism and glucoaldehyde acidification in phase Ⅱ metabolism. And there are related to the differences in the expression levels of phase Ⅰ and phase Ⅱ metabolic enzymes CYP2E1, CYP3A1, UGT1A1 and UGT1A6 in the liver.
2.Influencing factors for calcium salt deposition in patients with alveolar echinococcosis
Zitong XIONG ; Zhiyi LIN ; Yanxin HUANG ; Fuzhong FANG ; Zhengzhan WU ; Zirui XIN ; Chunxia HU ; Jiayu ZHOU ; Yuan YAO ; Hongwei ZHANG
Journal of Clinical Hepatology 2026;42(2):372-379
ObjectiveTo investigate the imaging features of calcium salt deposition and serological markers in patients with alveolar echinococcosis through a retrospective analysis, as well as independent risk factors for the degree of calcium salt deposition in lesions, and to provide a basis for assessing disease process. MethodsA retrospective analysis was performed for the imaging and clinical data of 107 patients with alveolar echinococcosis who were admitted to The First Affiliated Hospital of Shihezi University from December 2023 to June 2025, and according to the volume of calcium salt deposition, they were divided into non-deposition group with 16 patients, mild deposition group with 52 patients, moderate deposition group with 16 patients, and severe deposition group with 23 patients. A one-way analysis of variance or the Kruskal-Wallis H test was used for comparison of continuous data between groups, and the χ2 test or Fisher’s exact test was used for comparison of categorical data between groups. The four groups were further combined into the low deposition group (no/mild deposition) and the high deposition group (moderate/severe deposition). A binary logistic regression analysis was used to investigate the independent influencing factors for calcium salt deposition, and a predictive model was established. The receiver operating characteristic (ROC) curve was used to assess the predictive performance of the model, and the Bootstrap method was used for internal validation. ResultsThere were significant differences between the four groups in sex distribution, involvement of other sites, white blood cell count, lymphocyte percentage, fibrinogen, uric acid, sodium ion, chloride ion, and calcium ion (all P<0.05). The univariate analysis showed that there were significant differences between the four groups in sex, involvement of other sites, white blood cell count, lymphocyte percentage, fibrinogen, alanine aminotransferase, albumin, creatinine, uric acid, sodium ion, chloride ion, and calcium ion (all P<0.1). The multi-collinearity diagnosis showed that the VIF values for all continuous variables ranged from 1.104 to 1.760, suggesting that collinearity did not affect modeling. An ordinal logistic regression model was established based on sex, involvement of other sites, calcium ion, lymphocyte percentage, and uric acid. The multivariate analysis showed that lymphocyte percentage (odds ratio [OR]=1.106, 95% confidence interval [CI]: 1.041 — 1.174, P=0.001) and blood calcium level (OR=0.005, 95%CI: 0.000 —0.230, P=0.007) were independent influencing factors for the degree of calcium salt deposition. The regression equation was established as Logit(P)=8.231 + 0.100 × lymphocyte percentage -5.344 × calcium ion. The ROC curve analysis showed that the model had an area under the ROC curve of 0.716, with a Youden index of 0.353, a sensitivity of 1.000, and a specificity of 0.353. The Hosmer-Lemeshow test showed that the model had poor calibration (χ2=20.688, P=0.008). The Bootstrap method with 1000 repeated samples showed that the estimated values of lymphocyte percentage (OR=1.106, 95%CI: 1.049 — 1.186, P=0.002) and calcium ion (OR=0.005, 95%CI: 0.000 — 0.214, P=0.010) were consistent with the original model, and the confidence intervals did not include 1, which further supported the reliability of the model. ConclusionBoth lymphocyte percentage and blood calcium level are independent influencing factors for calcium salt deposition in alveolar echinococcosis, and the degree of calcium salt deposition in alveolar echinococcosis lesions increases with the reduction in blood calcium level and the increase in lymphocyte percentage.
3.Modified Xiaoyaosan Alleviates Neuronal Dysfunction in Rat Model of Post-myocardial Infarction Depression by Regulating Mitochondrial Quality Control Through Drp1/PINK1/Parkin Signaling Pathway
Zhen ZHONG ; Dongsheng WEI ; Xinyue XIONG ; Lin LI ; Mingli YAO ; Xinnuan SHI ; Youming JIANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):20-31
ObjectiveTo investigate the effects of modified Xiaoyaosan (JJXYS) on behavioral abnormalities and hippocampal mitochondrial quality control (MQC) in the rat model of post-myocardial infarction depression (PMD) and preliminarily explore its potential mechanism. MethodsA rat model of PMD was established by left anterior descending coronary artery ligation combined with chronic unpredictable mild stress (CUMS). Rats were randomized into a control group, a model group, a fluoxetine (FLX, 10 mg·kg-1) group, and low-, medium-, and high-dose JJXYS (JJXYS-L/M/H, 1.12, 2.24, 4.48 g·kg-1, respectively) groups. Depressive-like behaviors were evaluated by body weight monitoring, sucrose preference test, open field test, and forced swimming test. Hematoxylin-eosin staining and Nissl staining were used to observe hippocampal histomorphology and neuronal changes. Enzyme-linked immunosorbent assay was conducted to determine the serum levels of 5-hydroxytryptamine (5-HT), dopamine (DA), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). The mRNA levels of MQC-related genes including peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC-1α), nuclear respiratory factor 1 (Nrf1), and transcription factor A, mitochondrial (TFAM) in the hippocampal tissue were measured by real-time PCR. The expression of proteins related to the dynamin-related protein 1 (Drp1)/PTEN-induced putative kinase 1 (PINK1)/Parkin signaling pathway was determined by Western blot. ResultsCompared with the control group, the model group showed restricted body weight gain, aggravated depressive-like behaviors, declined serum 5-HT and DA levels, evident hippocampal neuronal damage and reduced Nissl bodies, as well as downregulated expression of MQC-related genes and proteins (P<0.05). Compared with the model group, both FLX and JJXYS alleviated the above changes to varying degrees. Moreover, the JJXYS-M and JJXYS-H groups showed more pronounced effects, improving behavioral performance, restoring 5-HT and DA levels, alleviating hippocampal pathological injury, and upregulating the expression of PGC-1α/Nrf1/TFAM mRNA and Drp1/PINK1/Parkin signaling pathway-related proteins (P<0.05). ConclusionJJXYS can significantly alleviate depressive-like behaviors and neurotransmitter imbalance in the rat model of PMD by regulating hippocampal MQC and upregulating the Drp1/PINK1/Parkin-related pathway. This study provides experimental evidence for the intervention of PMD with JJXYS.
4.Association of polychlorinated biphenyl exposure with platelet parameters across different glycemic states: The moderating role of a healthy lifestyle
Zhuo CHEN ; Huilin LOU ; Taimeng CHEN ; Fangyuan LIN ; Xueyan WU ; Yao GUO ; Haoran XU ; Mengke CHENG ; Peihan CHEN ; Yilin ZHOU ; Zhenxing MAO ; Xin TANG
Journal of Environmental and Occupational Medicine 2026;43(5):535-541
Background Platelet parameters are important indicators of cardiovascular risk, and environmental pollutants such as polychlorinated biphenyls (PCBs) may impair platelet function through oxidative stress. Objective To investigate the differential effects of single and mixed exposure to PCBs on platelet parameters among individuals with normal glucose tolerance (NGT), impaired fasting glucose (IFG), and type 2 diabetes mellitus (T2DM), and to evaluate the potential modifying role of a healthy lifestyle. Methods This study included 2249 participants (including 707 with NGT, 759 with IFG, and 783 with T2DM). Plasma PCB concentrations were measured using triple quadrupole gaschromatography-tandem mass spectrometry. Generalized linear regression was used to assess the associations between individual PCB congeners and platelet parameters. Quantile g-computation (QGC) and Bayesian kernel machine regression (BKMR) models were used to evaluate the overall effects of PCBs mixture exposure on platelet parameters across different glycemic states, as well as its interaction with healthy lifestyle score (HLS). Results Generalized linear regression analyses showed significant differences in the effects of PCBs on platelet parameters across different glycemic states (P<0.05). After adjusting for confounders, PCBs mixture exposure was significantly associated with lower platelet counts (PLT) in individuals with NGT (b=−10.60, 95%CI: −16.48, −4.71) and IFG (b=−12.91, 95%CI: −18.90, −6.92), whereas no significant association was observed in individuals with T2DM (P=0.051). Mean platelet volume (MPV) and platelet-large cell ratio (P-LCR) increased significantly with higher PCBs exposure levels across all three groups (P<0.05). BKMR analysis showed a positive association between PCBs mixture exposure and P-LCR, with the strongest association observed in the NGT group. Furthermore, a significant interaction was observed between HLS and PCBs mixture exposure, and a higher HLS attenuated the effects of PCBs on P-LCR. Conclusion Glycemic glycemic states may modify the effects of PCBs on platelets. Individuals with NGT appear more sensitive to PCBs exposure, whereas the T2DM state may attenuate this effect. Moreover, healthy lifestyles, including not smoking, moderate alcohol consumption, maintaining moderate-to-high physical activity, a healthy diet, and an appropriate body mass index (BMI), may mitigate the adverse effects of most PCBs on platelet parameters.
5.Association of polychlorinated biphenyl exposure with platelet parameters across different glycemic states: The moderating role of a healthy lifestyle
Zhuo CHEN ; Huilin LOU ; Taimeng CHEN ; Fangyuan LIN ; Xueyan WU ; Yao GUO ; Haoran XU ; Mengke CHENG ; Peihan CHEN ; Yilin ZHOU ; Zhenxing MAO ; Xin TANG
Journal of Environmental and Occupational Medicine 2026;43(5):535-541
Background Platelet parameters are important indicators of cardiovascular risk, and environmental pollutants such as polychlorinated biphenyls (PCBs) may impair platelet function through oxidative stress. Objective To investigate the differential effects of single and mixed exposure to PCBs on platelet parameters among individuals with normal glucose tolerance (NGT), impaired fasting glucose (IFG), and type 2 diabetes mellitus (T2DM), and to evaluate the potential modifying role of a healthy lifestyle. Methods This study included 2249 participants (including 707 with NGT, 759 with IFG, and 783 with T2DM). Plasma PCB concentrations were measured using triple quadrupole gaschromatography-tandem mass spectrometry. Generalized linear regression was used to assess the associations between individual PCB congeners and platelet parameters. Quantile g-computation (QGC) and Bayesian kernel machine regression (BKMR) models were used to evaluate the overall effects of PCBs mixture exposure on platelet parameters across different glycemic states, as well as its interaction with healthy lifestyle score (HLS). Results Generalized linear regression analyses showed significant differences in the effects of PCBs on platelet parameters across different glycemic states (P<0.05). After adjusting for confounders, PCBs mixture exposure was significantly associated with lower platelet counts (PLT) in individuals with NGT (b=−10.60, 95%CI: −16.48, −4.71) and IFG (b=−12.91, 95%CI: −18.90, −6.92), whereas no significant association was observed in individuals with T2DM (P=0.051). Mean platelet volume (MPV) and platelet-large cell ratio (P-LCR) increased significantly with higher PCBs exposure levels across all three groups (P<0.05). BKMR analysis showed a positive association between PCBs mixture exposure and P-LCR, with the strongest association observed in the NGT group. Furthermore, a significant interaction was observed between HLS and PCBs mixture exposure, and a higher HLS attenuated the effects of PCBs on P-LCR. Conclusion Glycemic glycemic states may modify the effects of PCBs on platelets. Individuals with NGT appear more sensitive to PCBs exposure, whereas the T2DM state may attenuate this effect. Moreover, healthy lifestyles, including not smoking, moderate alcohol consumption, maintaining moderate-to-high physical activity, a healthy diet, and an appropriate body mass index (BMI), may mitigate the adverse effects of most PCBs on platelet parameters.
6.Effect of heat-sensitive moxibustion at "Feishu" (BL13) on immunoinflammatory response in allergic rhinitis rats based on PI3K/AKT signaling pathway.
Yicheng LI ; Jun XIONG ; Meng LIN ; Han HU ; Lijun YAO
Chinese Acupuncture & Moxibustion 2025;45(7):957-966
OBJECTIVE:
To observe the effect of heat-sensitive moxibustion at "Feishu" (BL13) on immunoinflammatory response in rats with allergic rhinitis (AR) based on phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway, so as to explore its underlying mechanism.
METHODS:
Thirty-two male SD rats were randomly divided into a blank group (6 rats) and a modeling group (26 rats). In the modeling group, AR model was prepared using systemic and local attack sensitization method with ovalbumin. The successfully-modeled rats were randomized into a model group (6 rats), a medication group (6 rats) and a moxibustion group (14 rats). In the moxibustion group, the suspending moxibustion was operated at bilateral "Feishu" (BL13), 40 min each time, once daily, for 21 consecutive days; during which, the temperature of the body and tail was recorded. During intervention, if the temperature of the body and tail increased by >1 ℃, the heat-sensitive reaction at the point was determined in the rats of the moxibustion group, and these rats were collected in a heat-sensitive moxibustion group (8 rats involved and 6 rats of them were randomly collected to ensure the sample-size consistency); and those without heat-sensitive moxibustion reaction were assigned to a traditional moxibustion group (6 rats). In the medication group, fluticasone propionate nasal spray was applied, 8 μL on each side, once daily and for 21 days. The behavioral score for AR symptoms after modeling and intervention, and the content of serum immunoglobulin E (IgE) after modeling were observed. After intervention, the histological morphology of the nasal mucosa was observed using HE staining, the positive expression of thymic stromal lymphopoietin (TSLP) in the nasal mucosa was detected using immunohistochemistry, the levels of IgE, interleukin (IL)-4, IL-5, IL-13 and interferon-γ (IFN-γ) were detected by ELISA, and the protein expression of the member 4 of tumor necrosis factor receptor superfamily (OX40), phosphorylated protein kinase B (p-AKT), phosphorylated phosphatidylinositol 3-kinase (p-PI3K) in nasal mucosa was detected by Western blotting.
RESULTS:
After modeling, the behavioral score of AR symptoms and serum IgE level in the modeling group were higher than those of the blank group (P<0.01), suggesting the success of AR modeling. After intervention, compared with the blank group, the behavioral score of AR symptoms was increased (P<0.01);the nasal mucosa structure was disordered, the inflammatory infiltration was severe; the positive expression of TSLP in the nasal mucosa increased (P<0.01), the levels of serum IgE, IL-4, IL-5, and IL-13 elevated (P<0.01), and the level of IFN-γ decreased (P<0.01); and the protein expression of OX40, p-AKT, and p-PI3K in the nasal mucosa increased (P<0.05) in the model group. Compared with the model group, the behavioral score of AR symptoms was reduced (P<0.01); the nasal mucosa structure, inflammatory infiltration, and vascular dilation were ameliorated to varying degrees; the positive expression of TSLP in the nasal mucosa decreased (P<0.01); the content of serum IgE, IL-4, IL-5, and IL-13 decreased (P<0.05), and that of IFN-γ increased (P<0.05) in the medication, traditional moxibustion, and heat-sensitive moxibustion groups. Compared with the model group, the protein expression of p-AKT was reduced in the medication and traditional moxibustion groups (P<0.05), the protein expression of OX40, p-AKT, and p-PI3K in the nasal mucosa decreased in the heat-sensitive moxibustion group (P<0.05). When compared with the medication group, the positive expression of TSLP in the nasal mucosa was reduced (P<0.05) in the heat-sensitive moxibustion group. In comparison with the traditional moxibustion group, the content of serum IL-13 was reduced and the content of IFN-γ elevated in the heat-sensitive moxibustion and the medication groups (P<0.05), the protein expression of p-PI3K reduced in the medication group (P<0.05), and the positive expression of TSLP and the protein expression of OX40 and p-PI3K in the nasal mucosa were reduced in the heat-sensitive moxibustion group (P<0.05).
CONCLUSION
Heat-sensitive moxibustion at "Feishu" (BL13) can alleviate the symptoms of AR rats, ameliorate the inflammatory infiltration and telangiectasia of nasal mucosa, and inhibit immunoinflammatory response, which may be obtained by regulating PI3K/AKT signal pathway.
Animals
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Moxibustion
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Male
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Rats
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Signal Transduction
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Rats, Sprague-Dawley
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Rhinitis, Allergic/genetics*
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Proto-Oncogene Proteins c-akt/immunology*
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Acupuncture Points
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Humans
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Phosphatidylinositol 3-Kinases/immunology*
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Phosphatidylinositol 3-Kinase/immunology*
7.Summary of the academic thoughts of Professor XU Nenggui on the "governor vessel regulating spirit" acupuncture method for post-stroke dysphagia.
Hongtao LI ; Lubao FENG ; Lizhi ZHANG ; Lulu YAO ; Lin WANG ; Xu Nenggui DIRECTOR
Chinese Acupuncture & Moxibustion 2025;45(8):1128-1133
Based on his extensive clinical experience and the team's mechanistic research, Professor XU Nenggui has proposed the academic concept that "the governor vessel governs the brain and treats cerebral viscus diseases", and established a novel acupuncture approach for encephalopathy treatment centered on the integrated theory of "governor vessel-brain-mind", and developed a staged acupuncture protocol of "governor vessel regulating spirit" for ischemic stroke. This article introduces the academic features of this method in treating post-stroke dysphagia from four aspects: theoretical framework, treatment principles and point selection, mechanistic research, and clinical case studies. In clinical application, the method emphasizes syndrome differentiation based on meridians, harmonization between the conception and governor vessels; precise acupoint selection to treat both form and spirit; stage-specific differentiation with targeted needling. Furthermore, Professor XU integrates basic research with clinical practice, focusing on the neurobiological mechanisms underlying the efficacy of acupuncture in treating post-stroke dysphagia.
Acupuncture Therapy/methods*
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Humans
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Deglutition Disorders/psychology*
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Stroke/complications*
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Meridians
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Acupuncture Points
8."Outpatient+inpatient"bundled integrated payment on the utilization of health services for ordinary tuberculosis patients:An empirical analysis based on a city in Hebei Province
Peng-yu XU ; Yao YAO ; Lin-du ZHAO
Chinese Journal of Health Policy 2025;18(2):8-15
Objective:To examine the impact of the"outpatient+inpatient"bundled integrated payment reform on the utilization of health services for patients with ordinary tuberculosis,and to evaluate its effectiveness in resource allocation and cost containment.Methods:Based on the Andersen model,clinical and health insurance data from a city in Hebei Province were analyzed.An empirical study was conducted on 5 656 patients using Ordinary Least Squares(OLS)regression and Propensity Score Matching(PSM)to assess outpatient and inpatient service utilization and related costs.Results:Patients in the post-reform group showed a significant increase in outpatient visits and outpatient costs(an average increase of approximately 4.809 visits and 266.368 yuan,respectively,P<0.001),while inpatient visits,inpatient costs,and total medical costs significantly decreased(reductions of approximately 0.302 visits,3 638.647 yuan,and 3 372.279 yuan,respectively,P<0.001).Conclusion:The bundled integrated payment model effectively promotes outpatient service utilization,optimizes inpatient resource allocation,and reduces medical costs.However,the sharp increase in outpatient workload may pose a risk of overutilization.Recommendations:Future initiatives should establish a dynamic monitoring and evaluation system that emphasizes both service quality and clinical outcomes in outpatient and inpatient settings.Furthermore,refining incentive and regulatory mechanisms is essential to ensure that cost control measures do not compromise medical quality and patient safety.
9.Study on the mechanism of Jiawei Jisheng Shenqi Decoction regulating HIF-1α/Notch pathway to improve hypoxia and antagonize liver cirrhosis
Yiling MO ; Xiaoling ZHOU ; Lin LIU ; Dongqi SUN ; Teng WU ; Yi LUO ; Bowen RUAN ; Yueming WANG ; Yao JIA
Chinese Journal of Comparative Medicine 2025;35(2):1-12
Objective To explore the mechanism of Jiawei Jisheng Shenqi Decoction in improving the hypoxic microenvironment and antagonizing liver cirrhosis.Methods In vivo experiments were conducted using a rat model of carbon tetrachloride(CCL4)-induced liver cirrhosis.Rats were divided into normal,model,colchicine,JWJSSQ low-dose,JWJSSQ medium-dose,and JWJSSQ high-dose group.Pathological changes in liver tissues in each group were examined by hematoxylin and eosin(HE)and Masson staining,changes in serum liver function were detected using test kits,levels of hyaluronic acid(HA),laminin(LN),procollagen Ⅲ(PC Ⅲ),and collagen typeⅣ(COL4)were detected by enzyme-linked immunosorbent assay(ELISA),and protein expression levels of hypoxia-inducible factor-1α(HIF-1α),Notch1,Jagged1,and α-smooth muscle actin(α-SMA)were detected by Western blot.In vitro experiments were conducted in HSC-T6 cells,and the optimal concentration of CoCl2(100 μ mol/L,200μmol/L,400 μmol/L,600 μmol/L and 800 μmol/L)in the cultured cells and the optimal concentration of drug-containing serum(5%,10%,15%,20%)were determined by Cell Counting Kit-8(CCK-8)assay.The migration ability of cells in each group was detected by scratch testing,and changes in the apoptosis rates were determined by flow cytometry.Protein expression levels of HIF-1α,Notch1,Jagged1,α-SMA,matrix metallopeptidase 9(MMP9),and tissue inhibitor of metalloproteinases 1(TIMP-1)were detected by Western blot.Results In the in vivo experiments,liver swelling,inflammatory cell infiltration,collagen deposition,and the appearance of pseudolobules were significantly increased in the model group compared with those in the normal group.Serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),HA,LN,PCⅢ,and COL4 were significantly increased and albumin(ALB)was significantly decreased in the model group,while liver levels of HIF-1α,Notch1,Jagged1,and α-SMA proteins were significantly increased(P<0.01).Liver swelling,inflammatory cell infiltration,and collagen deposition were significantly reduced in each treatment group compared with those in the model group,and the degree of fibrosis was reduced.Serum ALT,AST,HA,LN,PCⅢ,and COL4 were significantly decreased and ALB was significantly increased,while liver levels of HIF-1α,Notch1,Jagged1,and α-SMA proteins were also significantly decreased to varying degrees(P<0.05).In the in vitro experiments,hypoxia promoted HSC-T6 migration and reduced apoptosis,increased the protein expression levels of HIF-1α,Notch1,Jagged1,α-SMA,and TIMP-1,and reduced the expression levels of MMP9(P<0.01).Serum containing Jiawei Jisheng Shenqi Decoction inhibited HSC-T6 migration,promoted HSC-T6 apoptosis,lowered the expression of HIF-1α,Notch1,Jagged1,α-SMA,and TIMP-1 proteins,and enhanced the expression of MMP9 protein(P<0.01).The inhibitory effect of Jiawei Jisheng Shenqi on HSC-T6 cell activation was reversed by the HIF-1α agonist dimethyloxalylglycine.Conclusions Jiawei Jisheng Shenqi Decoction can improve the hypoxic microenvironment via the HIF-1α/Notch pathway,thereby exerting an anti-liver cirrhosis effect.
10.Effects of forkhead box protein A1 knockout on microRNA expression profiles of benzoapyrene malignant transformed cells THBEc1
Zhiyu LIU ; Yujin FU ; Yitong LIN ; Juanling FU ; Biyun YAO ; Peng ZHAO
Chinese Journal of Pharmacology and Toxicology 2025;39(3):169-182
OBJECTIVE To explore the potential mechanisms of forkhead box protein A1(FOXA1)in benzo[a]pyrene(BaP)-induced carcinogenesis by investigating the effect of FOXA1 by knockout on microRNA(miRNA)expression profiles in BaP malignant transformed cells THBEc1 and establishing regulatory networks between FOXA1,miRNA and their target genes.METHODS FOXA1 knockout THBEc1 cells THBEc1-ΔFOXA1-c34 and control cells THBEc1-ctrl were used as study models.Western blotting was employed to determine FOXA1 protein expression levels.Next-generation sequencing(NGS)tech-nology was used to identify differentially expressed miRNAs between THBEc1-ΔFOXA1-c34 and THBEc1-ctrl cells,with subsequent validation by RT-qPCR.Five databases(ENCORI,miRDB,mirDIP,miRWalk and TargetScan 8.0)were used in conjunction with NGS results of mRNA between THBEc1-ΔFOXA1-c34 and THBEc1-ctrl to predict different expressed genes(DEGs)regulated by the identified differentially expressed miRNAs.GO and KEGG enrichment analyses were conducted on the DEGs using the DAVID database.Interaction network analysis of the proteins encoded by the DEGs was performed using STRING 12.0 and Cytoscape 3.10.2 software.RESULTS No FOXA1 expression was detected in THBEc1-ΔFOXA1-c34 cells.A differential analysis of miRNA expressions revealed 33 miRNAs with a fold change of>2 or<0.5 and a false discovery rate of<0.05 between THBEc1-ΔFOXA1-c34 and THBEc1-ctrl cells,13 of which were down-regulated and 20 were up-regulated in THBEc1-ΔFOXA1-c34 cells.A regulatory network was formed by 11 down-regulated miRNAs and 32 up-regulated mRNAs,while a second network included 16 up-regulated miRNAs and 56 down-regulated mRNAs.The 27 differentially expressed miRNAs participated in various biological processes through the regulation of 88 DEGs,primarily associated with cell growth,proliferation,migration,apoptosis,angiogenesis,epithe-lial-mesenchymal transition,and signal transduction(TGF-β,Hippo,NF-kappa B and MAPK pathways).CONCLUSION The miRNA expression profile in BaP-malignant transformed THBEc1 cells is altered following FOXA1 knockout that may disrupt TGF-β and MAPK signaling pathways by changing miRNA expression levels,thereby inhibiting cell proliferation and migration.

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